Sample records for plants assay purification

  1. Phytochrome from green plants: Assay, purification, and characterization

    Energy Technology Data Exchange (ETDEWEB)

    Quail, P.H. (California Univ., Berkeley, CA (United States). Dept. of Plant and Soil Biology Agricultural Research Service, Albany, CA (United States). Plant Gene Expression Center)


    This funding period was directed at developing an in-depth molecular analysis of the low-abundance, 118,000 M{sub r} green-tissue phytochrome that had at that time been relatively recently identified as being distinct from the better characterized 124,000 M{sub r} phytochrome abundant in etiolated tissue. The specific objectives as stated in the original proposal were: (1) To generate monoclonal antibodies specific to the 118,000 M{sub r} green-tissue phytochrome. (2) To develop additional and improved procedures to permit progress toward the ultimate goal of purifying green-tissue phytochrome to homogeneity. (3) To initiate an alternative approach to determining the structural properties of green-tissue phytochrome by isolating and sequencing cDNA cones representing the 118,000 M{sub r} green-tissue polypeptide in Avena. This approach is based on and will test hypothesis that the 118,000 M{sub r} polypeptide is encoded by a gene(s) distinct from those encoding etiolated-tissue 124,000 M{sub r} phytochrome. (4) To utilize any such 118,000 M{sub r} phytochrome specific cDNA clones as hybridization probes to begin to investigate the structure, organization, and regulation of the corresponding gene(s) in Avena. (5) To begin to investigate the possible presence in other higher plant and algal species of sequences homologous to the 118,000 M{sub r} Avena polypeptide using the Avena clones at hybridization probes. Most of these objectives have been accomplished, at least in principle, although the major breakthrough establishing that phytochrome is encoded by a multigene family came from the use of Arabidopsis rather than Avena. Similarly, much of the characterization subsequent to this discovery has been performed in Arabidopsis and rise as model dicot and monocot systems, respectively, rather than Avena. 13 refs., 9 figs.

  2. Isolation, production, purification, assay and characterization of ...

    African Journals Online (AJOL)

    Isolation, production, purification, assay and characterization of fibrinolytic ... are isolated from Bacillus subtilis, β-haemolytic Streptococci and urine sample. ... recombinant E.coli containing short fragment genomic DNA of Pseudomonas sp.

  3. Purification of Water by Aquatic Plants


    Morimitsu, Katsuhito; Kawahigashi, Tatsuo


    [Abstract] Water quality purification of many water systems including those occurring in rivers depends to a great degree on water quality purification activities of aquatic plants and microbes. This paper presents a discussion of results, based on laboratory experiments, of purification by aquatic plants.

  4. 植物几丁质酶纯化、测定及应用研究进展%Review on purification, enzyme assay and application of plant chitinases

    Institute of Scientific and Technical Information of China (English)

    杨海霞; 邓建军; 张建; 赵广华


    植物几丁质酶是植物体中能够水解几丁质多聚体的一种致病性相关蛋白(Pathogenesis-related proteins).近年来对于几丁质酶的研究报道中,大量新型的植物几丁质酶被分离纯化,并建立了不同的酶活测定方法,在几丁质酶的结构及分类方面也逐步有了系统的研究.从几丁质酶的结构及分类,分离纯化以及已建立的酶活测定方法等方面取得的新进展进行了综述,并展望了几丁质酶在农业、食品生产及药用领域的应用前景.%Plant chitinases which hydrolyze the chitin is one of pathogenesis-related proteins and can be induced in resistance of plants to fungal pathogens. Thus, plant chitinases has a wide range of application as a antisepticise material. Recently, chitinases from different plants have been purified and their enzymatic activities have been assayed with varied methods. The structures and classifications, different methods on enzymatic activity assay and purification were summarized. Moreover, applications of chitinases such as in food and medicine field were prospected.

  5. Ionic behavior of treated water at a water purification plant


    Yanagida, Kazumi; Kawahigashi, Tatsuo


    [Abstract] Water at each processing stage in a water purification plant was extracted and analyzed to investigate changes of water quality. Investigations of water at each processing stage at the water purification plant are discussed herein.

  6. Ionic behavior of treated water at a water purification plant


    Yanagida, Kazumi; Kawahigashi, Tatsuo


    [Abstract] Water at each processing stage in a water purification plant was extracted and analyzed to investigate changes of water quality. Investigations of water at each processing stage at the water purification plant are discussed herein.

  7. Purification and kinase assay of PKN. (United States)

    Mukai, Hideyuki; Ono, Yoshitaka


    PKN is a serine/threonine protein kinase, which has a catalytic domain highly homologous to that of protein kinase C (PKC) in the carboxyl-terminal region and three repeats of the antiparallel coiled coil (ACC) domain in the amino-terminal region. Mammalian PKN has three isoforms each derived from different genes, PKN1 (PKNalpha/PRK1/PAK1), PKN2 (PRK2/PAK2/PKNgamma), and PKN3 (PKNbeta). PKN isoforms show different enzymatic properties and tissue distributions and have been implicated in various distinct cellular processes (reviewed in Mukai [2003]). This chapter discusses methods to prepare purified enzymes and to assay substrate phosphorylation activities.

  8. [Human angiogenin: expression, purification, biological assay]. (United States)

    Yang, H; Zhang, Y Q; Yan, Z; Han, W; Yao, L B; Su, C Z


    Angiogenin cDNA was obtained by RT-PCR, and cloned into the fusion expression vector pRSETB. The recombinant Angiogenin protein was fused with His6 at its N-terminal and expressed as inclusion body. The expression level was about 10% of the total bacteria protein. After dissolved in 8 mol/L urea, the recombinant protein was purified by Ni2(+)-NTA chelating resin, according to the high affinity of His6 with Ni2+. The biological assay indicated that purified rhANG could induced the new blood vessel formation of CAM and degraded tRNA in vitro.

  9. Direct LAMP Assay without Prior DNA Purification for Sex Determination of Papaya

    Directory of Open Access Journals (Sweden)

    Chi-Chu Tsai


    Full Text Available Papaya (Carica papaya L. is an economically important tropical fruit tree with hermaphrodite, male and female sex types. Hermaphroditic plants are the major type used for papaya production because their fruits have more commercial advantages than those of female plants. Sex determination of the seedlings, or during the early growth stages, is very important for the papaya seedling industry. Thus far, the only method for determining the sex type of a papaya at the seedling stage has been DNA analysis. In this study, a molecular technique—based on DNA analysis—was developed for detecting male-hermaphrodite-specific markers to examine the papaya’s sex type. This method is based on the loop-mediated isothermal amplification (LAMP and does not require prior DNA purification. The results show that the method is an easy, efficient, and inexpensive way to determine a papaya’s sex. This is the first report on the LAMP assay, using intact plant materials-without DNA purification-as samples for the analysis of sex determination of papaya. We found that using high-efficiency DNA polymerase was essential for successful DNA amplification, using trace intact plant material as a template DNA source.

  10. Direct LAMP Assay without Prior DNA Purification for Sex Determination of Papaya. (United States)

    Tsai, Chi-Chu; Shih, Huei-Chuan; Ko, Ya-Zhu; Wang, Ren-Huang; Li, Shu-Ju; Chiang, Yu-Chung


    Papaya (Carica papaya L.) is an economically important tropical fruit tree with hermaphrodite, male and female sex types. Hermaphroditic plants are the major type used for papaya production because their fruits have more commercial advantages than those of female plants. Sex determination of the seedlings, or during the early growth stages, is very important for the papaya seedling industry. Thus far, the only method for determining the sex type of a papaya at the seedling stage has been DNA analysis. In this study, a molecular technique-based on DNA analysis-was developed for detecting male-hermaphrodite-specific markers to examine the papaya's sex type. This method is based on the loop-mediated isothermal amplification (LAMP) and does not require prior DNA purification. The results show that the method is an easy, efficient, and inexpensive way to determine a papaya's sex. This is the first report on the LAMP assay, using intact plant materials-without DNA purification-as samples for the analysis of sex determination of papaya. We found that using high-efficiency DNA polymerase was essential for successful DNA amplification, using trace intact plant material as a template DNA source.

  11. A Cell Lysis and Protein Purification - Single Molecule Assay Devices for Evaluation of Genetically Engineered Proteins (United States)

    Nakyama, Tetsuya; Tabata, Kazuhito; Noji, Hiroyuki; Yokokawa, Ryuji

    We have developed two devices applicable to evaluate genetically engineered proteins in single molecule assay: on-chip cell lysis device, and protein purification - assay device. A motor protein, F1-ATPase expressed in E.coli, was focused in this report as a target protein. Cell lysis was simply performed by applying pulse voltage between Au electrodes patterned by photolithography, and its efficiency was determined by absorptiometry. The subsequent processes, purification and assay of extracted proteins, were demonstrated in order to detect F1-ATPase and to evaluate its activity. The specific bonding between his-tag in F1-ATPase and Ni-NTA coated on a glass surface was utilized for the purification process. After immobilization of F1-ATPase, avidin-coated microspheres and adenosine tri-phosphate (ATP) solution were infused sequentially to assay the protein. Microsphere rotation was realized by activity of F1-ATPase corresponding to ATP hydrolysis. Results show that the cell lysis device, at the optimum condition, extracts enough amount of protein for single molecule assay. Once cell lysate was injected to the purification - assay device, proteins were diffused in the lateral direction in a Y-shape microchannel. The gradient of protein concentratioin provides an optimal concentration for the assay i.e. the highest density of rotating beads. Density of rotating beads is also affected by the initial concentration of protein injected to the device. The optimum concentration was achieved by our cell lysis device not by the conventional method by ultrasonic wave. Rotation speed was analyzed for several microspheres assayed in the purification - assay device, and the results were compatible to that of conventional assay in which F1-ATPase was purified in bulk scale. In conclusion, we have demonstrated on-chip cell lysis and assay appropriate for the sequential analysis without any pretreatment. On-chip devices replacing conventional bioanalytical methods will be

  12. Plant lipases: partial purification of Carica papaya lipase. (United States)

    Rivera, Ivanna; Mateos-Díaz, Juan Carlos; Sandoval, Georgina


    Lipases from plants have very interesting features for application in different fields. This chapter provides an overview on some of the most important aspects of plant lipases, such as sources, applications, physiological functions, and specificities. Lipases from laticifers and particularly Carica papaya lipase (CPL) have emerged as a versatile autoimmobilized biocatalyst. However, to get a better understanding of CPL biocatalytic properties, the isolation and purification of individual C. papaya lipolytic enzymes become necessary. In this chapter, a practical protocol for partial purification of the latex-associated lipolytic activity from C. papaya is given.

  13. A Scintillator Purification Plant and Fluid Handling System for SNO+

    CERN Document Server

    Ford, Richard J


    A large capacity purification plant and fluid handling system has been constructed for the SNO+ neutrino and double-beta decay experiment, located 6800 feet underground at SNOLAB, Canada. SNO+ is a refurbishment of the SNO detector to fill the acrylic vessel with liquid scintillator based on Linear Alkylbenzene (LAB) and 2 g/L PPO, and also has a phase to load natural tellurium into the scintillator for a double-beta decay experiment with 130Te. The plant includes processes multi-stage dual-stream distillation, column water extraction, steam stripping, and functionalized silica gel adsorption columns. The plant also includes systems for preparing the scintillator with PPO and metal-loading the scintillator for double-beta decay exposure. We review the basis of design, the purification principles, specifications for the plant, and the construction and installations. The construction and commissioning status is updated.

  14. Indoor Air Purification by Potted Plants

    DEFF Research Database (Denmark)

    Dela Cruz, Majbrit

    Volatile organic compounds (VOC) are ubiquitous in the indoor environment and can affect human health negatively. Potted plants are a potential green technology solution for removal of VOCs. This PhD project aimed at reviewing current literature on VOC removal by potted plants, developing a dynam...... community structure was, however, not the same for two consecutive experiments indicating that several bacteria can degrade VOCs...

  15. [The extraction, purification and assaying of Gynostemma pentaphyllum polysaccharides]. (United States)

    Song, Shu-liang; Tang, Jin-bao; Ji, Ai-guo; Liang, Hao; Zhu, Peng; Wang, Wei-li


    By orthogonal design, and considering extracting efficiency and cost, optimizing the extract method of Gynostemma pentaphyllum polysaccharides. We purified the crude Gynostemma pentaphyllum polysaccharides initially, and assayed the polysaccharides content of Gynostemma pentaphyllum polysccharides. The content of Gynostemma pentaphyllum polysaccharides was sigificantly higher than the predecessor. It would provide conditions for the deep exploitation of Gynostemma pentaphyllum.

  16. Turnkey Helium Purification and Liquefaction Plant for DARWIN, Australia (United States)

    Lindemann, U.; Boeck, S.; Blum, L.; Kurtcuoglu, K.


    The Linde Group, through its Australian subsidiary BOC Limited, has signed an agreement with Darwin LNG Pty Ltd for the supply of feed-gas to Linde's new helium refining and liquefaction facility in Darwin, Australia. Linde Kryotechnik AG, located in Switzerland, has carried out the engineering and fabrication of the equipment for the turn key helium plant. The raw feed gas flow of 20'730 Nm3/h contains up to of 3 mol% helium. The purification process of the feed gas consists of partial condensation of nitrogen in two stages, cryogenic adsorption and finally catalytic oxidation of hydrogen followed by a dryer system. Downstream of the purification the refined helium is liquefied using a modified Bryton process and stored in a 30'000 gal LHe tank. For further distribution and export of the liquid helium there are two stations available for filling of truck trailers and containers. The liquid nitrogen, required for refrigeration capacity to the nitrogen removal stages in the purification process as well as for the pre-cooling of the pure helium in the liquefaction process, is generated on site during the feed gas purification process. The optimized process provides low power consumption, maximum helium recovery and a minimum helium loss.

  17. Single-Step Purification and Characterization of A Recombinant Serine Proteinase Inhibitor from Transgenic Plants. (United States)

    Jha, Shweta; Agarwal, Saurabh; Sanyal, Indraneel; Amla, D V


    Expression of recombinant therapeutic proteins in transgenic plants has a tremendous impact on safe and economical production of biomolecules for biopharmaceutical industry. The major limitation in their production is downstream processing of recombinant protein to obtain higher yield and purity of the final product. In this study, a simple and rapid process has been developed for purification of therapeutic recombinant α1-proteinase inhibitor (rα1-PI) from transgenic tomato plants, which is an abundant serine protease inhibitor in human serum and chiefly inhibits the activity of neutrophil elastase in lungs. We have expressed rα1-PI with modified synthetic gene in transgenic tomato plants at a very high level (≃3.2 % of total soluble protein). The heterologous protein was extracted with (NH4)2SO4 precipitation, followed by chromatographic separation on different matrices. However, only immunoaffinity chromatography resulted into homogenous preparation of rα1-PI with 54 % recovery. The plant-purified rα1-PI showed molecular mass and structural conformation comparable to native serum α1-PI, as shown by mass spectrometry and optical spectroscopy. The results of elastase inhibition assay revealed biological activity of the purified rα1-PI protein. This work demonstrates a simple and efficient one-step purification of rα1-PI from transgenic plants, which is an essential prerequisite for further therapeutic development.

  18. Recombinant Plasmodium falciparum glutamate rich protein; purification and use in enzyme-linked immunosorbent assay

    DEFF Research Database (Denmark)

    Dziegiel, M; Borre, Mette; Jepsen, S


    A method for purification of a recombinant Plasmodium falciparum protein produced in E. coli and its use in an enzyme-linked immunosorbent assay (ELISA) is described. The cloned gene fragment encodes GLURP,489-1271 the carboxy-terminal 783 amino acid residue portion of a 1271 amino acid residue P...... of the immunogenicity of a possible future P. falciparum vaccine utilizing epitopes from GLURP....

  19. Expression and affinity purification of recombinant proteins from plants (United States)

    Desai, Urvee A.; Sur, Gargi; Daunert, Sylvia; Babbitt, Ruth; Li, Qingshun


    With recent advances in plant biotechnology, transgenic plants have been targeted as an inexpensive means for the mass production of proteins for biopharmaceutical and industrial uses. However, the current plant purification techniques lack a generally applicable, economic, large-scale strategy. In this study, we demonstrate the purification of a model protein, beta-glucuronidase (GUS), by employing the protein calmodulin (CaM) as an affinity tag. In the proposed system, CaM is fused to GUS. In the presence of calcium, the calmodulin fusion protein binds specifically to a phenothiazine-modified surface of an affinity column. When calcium is removed with a complexing agent, e.g., EDTA, calmodulin undergoes a conformational change allowing the dissociation of the calmodulin-phenothiazine complex and, therefore, permitting the elution of the GUS-CaM fusion protein. The advantages of this approach are the fast, efficient, and economical isolation of the target protein under mild elution conditions, thus preserving the activity of the target protein. Two types of transformation methods were used in this study, namely, the Agrobacterium-mediated system and the viral-vector-mediated transformation system. Copyright 2002 Elsevier Science (USA).

  20. [Protoplasts isolation, purification and plant regeneration of Pinellia cordata]. (United States)

    Yang, Xian; Ma, Dan-Dan; Jiang, Fu-Sheng; Chen, Ni-Pi; Ding, Bin; Jin, Li-Xia; Qian, Chao-Dong; Ding, Zhi-Shan


    The main factors which affected the isolation, purification and cultivation of Pinellia cordata protoplasts from leaves were studied. The results indicated that the optimum enzyme solution for P. cordata leaves was 13% CPW + 1.0% Cellulose +0.1% Pectolase, at pH 6.0, temperature (25-28 degrees C ) for 4 h. The sucrose density gradient centrifugation was adopted to purificate the protoplasts collected, when 25% sucrose was used as mediator, centrifugating at 500 rpm for 10 min. When the protoplasts were shallow liquid and liquid-solid double layer cultured on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA + 13% mannitol at the density of 2.5 x 104 protoplasts/mL, or fed and nursed cultured at the density of 100-500 protoplasts/mL, cell division could be observed for 3 days; granular calli appeared for 30 days. Calli was proliferated on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA solidified by 0.55% agar, and differentiated and regenerated after 5-6 months. Plant generation of P. cordata is successfully established.

  1. A highly sensitive cell assay for validation of purification regimes of alginates. (United States)

    Leinfelder, U; Brunnenmeier, F; Cramer, H; Schiller, J; Arnold, K; Vásquez, J A; Zimmermann, U


    Among the hydrogels used for microencapsulation of cells and tissues, alginate has been and will continue to be one of the most important biomaterials. A mandatory requirement for clinical immunoisolated transplantations is the reproducible production of biocompatible alginate. As shown here for alginates extracted from freshly collected algal stipes, the current assays used for validation of the quality of the alginate are not sufficient to screen for impurities arising from spores of gram-positive bacteria (and related contaminants). To assess the quality of alginate, we have developed a cell assay based on the induction of apoptosis in Jurkat cells. This assay allows in combination with the "modified mixed lymphocyte" assay a rapid and sensitive screening for any fibrosis-inducing impurities in alginate samples (even during the purification regime) as demonstrated by transplantation experiments performed in parallel with BB rats (exhibiting an elevated macrophage activity). The results clearly demonstrate that the quality of the input algal material is of key relevance for the production of transplantation-grade alginate.

  2. A sialic acid assay in isolation and purification of bovine k-casein glycomacropeptide: a review. (United States)

    Nakano, Takuo; Ozimek, Lech


    Sialic acid is a carbohydrate moiety of k-casein glycomacropeptide (GMP), which is a 64 amino acid residue C-terminal sialylated phosphorylated glycopeptide released from k-casein by the action of chymosin during cheese making. GMP lacks aromatic amino acids including phenylalanine, tyrosine, and tryptophan. Because of its unique amino acid composition and various biological activities, GMP is thought to be a potential ingredient for dietetic foods (e.g., a food for PKU patients) and pharmaceuticals. Thus, increased attention has been given to the development of techniques to purify GMP. In this review, techniques of GMP purification described in patents and scientific research papers were introduced. A sialic acid assay is the important method to track GMP isolation and purification processes, for which the thiobarbituric acid reaction with 1-propanol as a chromophore extracting solvent is an inexpensive, practical and specific technique. Sephacryl S-200 gel filtration chromatography, cellulose acetate electrophoresis, and sodium dodecyl sulfate polyacrylamide gel electrophoresis are the major techniques to identify sialic acid specific to GMP. Sephacryl S-200 chromatography and cellulose acetate electrophoresis are also used to detect GMP sialic acid in whey pearmeate and whey added commercial margarine samples. Future research includes development of an economical industrial scale method to produce high purity GMP.

  3. A simple purification and activity assay of the coagulant protein from Moringa oleifera seed. (United States)

    Ghebremichael, Kebreab A; Gunaratna, K R; Henriksson, Hongbin; Brumer, Harry; Dalhammar, Gunnel


    Use of extracts from Moringa oleifera (MO) is of great interest for low-cost water treatment. This paper discusses water and salt extraction of a coagulant protein from the seed, purification using ion exchange, its chemical characteristics, coagulation and antimicrobial properties. The coagulant from both extracts is a cationic protein with pI greater than 9.6 and molecular mass less than 6.5 kDa. Mass spectrometric analysis of the purified water extract indicated that it contained at least four homologous proteins, based on MS/MS peptide sequence data. The protein is thermoresistant and remained active after 5h heat treatment at 95 degrees C. The coagulant protein showed both flocculating and antibacterial effects of 1.1--4 log reduction. With samples of high turbidity, the MO extract showed similar coagulation activity as alum. Cecropin A and MO extract were found to have similar flocculation effects for clay and microorganisms. Simple methods for both the purification and assay of MO coagulating proteins are presented, which are necessary for large-scale water treatment applications.

  4. Electrophoretic purification of radioiodinated follicle-stimulating hormone for radioligand receptor assay and radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Schneyer, A.L.; Sluss, P.M.; Bosukonda, D.; Reichert, L.E. Jr.


    A method is described for electrophoretic purification of (/sup 125/I)human (h) FSH after radioiodination that improves radioligand binding to FSH membrane receptors. Lactoperoxidase-iodinated hFSH was separated from reaction products by electrophoresis on 7.5% polyacrylamide tube gels (PAGE). Material eluted from 3-mm gel slices was analyzed for incorporation of /sup 125/I and binding to antibody (RIA) or receptor (RRA), and by sodium dodecyl sulfate-PAGE for protein composition. Sodium dodecyl sulfate-PAGE analysis of individual PAGE fractions demonstrated that iodinated proteins, both higher and lower in apparent mol wt than intact FSH, were separated by PAGE, but not by gel filtration chromatography (Sephadex G-25). PAGE purification of radioligand resulted in significantly greater (compared to gel filtration) RRA sensitivity and specificity. Maximum binding of PAGE-purified (/sup 125/I)hFSH to excess calf tests membrane receptors was 45%, with a specific activity of approximately 26 microCi/micrograms, as determined by the method of self-displacement. Maximum binding to excess hFSH antisera (NIH anti-hFSH 4) was 80-85%. This allowed a useful final dilution of 1:120,000, thereby facilitating development of a sensitive and specific RIA with this antiserum. These data indicate that PAGE separation of intact (/sup 125/I)hFSH from other iodinated proteins results in improved radioligand binding, assay sensitivity, and assay specificity. In addition, PAGE-purified lactoperoxidase-iodinated hFSH is suitable for use in both RIA and RRA.

  5. Fungicide resistance assays for fungal plant pathogens. (United States)

    Secor, Gary A; Rivera, Viviana V


    Fungicide resistance assays are useful to determine if a fungal pathogen has developed resistance to a fungicide used to manage the disease it causes. Laboratory assays are used to determine loss of sensitivity, or resistance, to a fungicide and can explain fungicide failures and for developing successful fungicide recommendations in the field. Laboratory assays for fungicide resistance are conducted by measuring reductions in growth or spore germination of fungi in the presence of fungicide, or by molecular procedures. This chapter describes two techniques for measuring fungicide resistance, using the sugarbeet leaf spot fungus Cercospora beticola as a model for the protocol. Two procedures are described for fungicides from two different classes; growth reduction for triazole (sterol demethylation inhibitor; DMI) fungicides, and inhibition of spore germination for quinone outside inhibitor (QoI) fungicides.

  6. Extraction and purification methods in downstream processing of plant-based recombinant proteins. (United States)

    Łojewska, Ewelina; Kowalczyk, Tomasz; Olejniczak, Szymon; Sakowicz, Tomasz


    During the last two decades, the production of recombinant proteins in plant systems has been receiving increased attention. Currently, proteins are considered as the most important biopharmaceuticals. However, high costs and problems with scaling up the purification and isolation processes make the production of plant-based recombinant proteins a challenging task. This paper presents a summary of the information regarding the downstream processing in plant systems and provides a comprehensible overview of its key steps, such as extraction and purification. To highlight the recent progress, mainly new developments in the downstream technology have been chosen. Furthermore, besides most popular techniques, alternative methods have been described.

  7. Presence of Acanthamoeba water purification plants in southern England

    Institute of Scientific and Technical Information of China (English)

    Shanmuganathan V; Khan NA


    Objective:To identify the prevalence of Acanthamoeba in drinking water treatment plants during the course of the purification processes.Methods:Samples were taken from two drinking water purification plants and moni-tored for the presence of Acanthamoeba in order to estimate the removal capacity of treatment methods em-ployed.Water samples were collected at each step in the purification,during the one year survey,and ana-lysed for the presence of Acanthamoeba plating on bacterial-seeded plates.Results:The results showed that amoebae were present in surface raw waters in 100 % of the samples tested.Acanthamoeba spp.were iso-lated from 71 % and 57 % of the water samples collected from post flat-bottom clarifier 1 and post-sedimenta-tion plant respectively.Considering the outflow drinking waters,the removal capacity was 100 % in both puri-fication plants monitored.The occurrence of Acanthamoeba was not associated with seasonality.Conclusion:These findings confirm that water purification plants employing methods of flocculation,sedimentation,and fil-tration in combination with activated charcoal filtration,ozonisation and chlorination exhibited sufficient Acan-thamoeba removal capacity and the presence of amoebae in the tap water may be due to older plumbing,water storage tanks,tap water hygiene,and /or environmental settings.

  8. The SNO+ Scintillator Purification Plant and Projected Sensitivity to Solar Neutrinos in the Pure Scintillator Phase (United States)

    Pershing, Teal; SNO+ Collaboration


    The SNO+ detector is a neutrino and neutrinoless double-beta decay experiment utilizing the renovated SNO detector. In the second phase of operation, the SNO+ detector will contain 780 tons of organic liquid scintillator composed of 2 g/L 2,5-diphenyloxazole (PPO) in linear alkylbenzene (LAB). In this phase, SNO+ will strive to detect solar neutrinos in the sub-MeV range, including CNO production neutrinos and pp production neutrinos. To achieve the necessary detector sensitivity, a four-part scintillator purification plant has been constructed in SNOLAB for the removal of ionic and radioactive impurities. We present an overview of the SNO+ scintillator purification plant stages, including distillation, water extraction, gas stripping, and metal scavenger columns. We also give the projected SNO+ sensitivities to various solar-produced neutrinos based on the scintillator plant's projected purification efficiency.

  9. The use of comet assay in plant toxicology: recent advances

    Directory of Open Access Journals (Sweden)

    Conceição LV Santos


    Full Text Available The systematic study of genotoxicity in plants induced by contaminants and other stress agents has been hindered to date by the lack of reliable and robust biomarkers. The comet assay is a versatile and sensitive method for the evaluation of DNA damages and DNA repair capacity at single-cell level. Due to its simplicity and sensitivity, and the small number of cells required to obtain robust results, the use of plant comet assay has drastically increased in the last decade. For years its use was restricted to a few model species, e.g. Allium cepa, Nicotiana tabacum, Vicia faba, or Arabidopsis thaliana but this number largely increased in the last years. Plant comet assay has been used to study the genotoxic impact of radiation, chemicals including pesticides, phytocompounds, heavy metals, nanoparticles or contaminated complex matrices. Here we will review the most recent data on the use of this technique as a standard approach for studying the genotoxic effects of different stress conditions on plants. Also, we will discuss the integration of information provided by the comet assay with other DNA-damage indicators, and with cellular responses including oxidative stress, cell division or cell death. Finally, we will focus on putative relations between transcripts related with DNA damage pathways, DNA replication and repair, oxidative stress and cell cycle progression that have been identified in plant cells with comet assays demonstrating DNA damage.

  10. Large-scale purification of synaptophysin and quantification with a newly established enzyme-linked immunosorbent assay. (United States)

    Schlaf, G; Göddecke, M; Wolff, J R; Felgenhauer, K; Mäder, M


    Synaptophysin (SYP I), an integral membrane protein, was purified on a large scale (0.55 - 2.7 mg) from isolated small synaptic vesicles (SSV) of porcine cortex. In order to achieve this, a conventional purification procedure which consists of size exlusion chromatography, hydrophobic interaction chromatography and chromatofocusing has been developed. This procedure was compared with purification of SYP I by immunoaffinity chromatography. The elution patterns of both procedures were monitored using sodium dodecylsulfate gel electrophoresis (SDS-PAGE) with subsequent Coomassie blue staining of proteins and simultaneous immunoblotting with SYP I-specific antibody. Contaminating proteins with relative molecular masses (M(r)) very similar to SYP I could be removed during the process of purification, demonstrating that the 38 kDa protein found after Triton X-100 lysis of enriched SSV does not exclusively represent SYP I. A specific antiserum was raised in rabbits using a highly purified preparation of SYP I. This antiserum was used in combination with a monoclonal antibody to establish a specific and sensitive enzyme-linked immunosorbent assay (ELISA) which allowed rapid and reliable quantification of this hydrophobic membrane protein in all purification steps, starting with Triton X-100-lysed brain homogenates. Using this ELISA, the concentration of SYP I in highly purified SSV was determined to be 5.8% of solubilized protein.

  11. Valorization technics by means of vermiculture for fatty wastes resulting from wastes water purification plants

    Energy Technology Data Exchange (ETDEWEB)

    Vignoles, C. (Service Assainissement, 31 - Toulouse (France))

    Fats, scums and other floating organic wastes extracted from waste water purification plants have always caused important problems of treatment to specialists. Municipal and technical services of Toulouse have elaborated an original valorization process. Results are simultaneously spectacular for environment and economically reasonable. One may think that this natural method is bound to experience interesting developments in the future.

  12. Utilization of red mud for the purification of waste waters from nuclear power plants

    Energy Technology Data Exchange (ETDEWEB)

    Luka, Mikelic; Visnja, Orescanin; Stipe, Lulic [Rudjer Boskovic Institute, Lab. for radioecology, Zagreb (Croatia)


    Sorption of the radionuclides and heavy metals from low level liquid radioactive waste on the coagulant produced from bauxite waste (red mud and waste base) was presented. Research was conducted on composite annual samples of waste water collected in the Waste Monitor Tank (W.M.T.) from Kro Nuclear Power Plant during each month. Activities of radionuclide in W.M.T. were measured before and after purification using high purity germanium detector. Also, elemental concentrations in W.M.T. before and after purification were measured by source excited energy dispersive X-ray fluorescence (E.D.X.R.F.). It has been showed that activated red mud is excellent purification agent for the removal of radionuclides present in low level liquid radioactive waste. Removal efficiency was 100% for the radionuclides {sup 58}Co and {sup 60}Co 100%, and over 60% for {sup 134}Cs and {sup 137}Cs. (authors)

  13. Enzymatic assay for calmodulins based on plant NAD kinase activity

    Energy Technology Data Exchange (ETDEWEB)

    Harmon, A.C.; Jarrett, H.W.; Cormier, M.J.


    NAD kinase with increased sensitivity to calmodulin was purified from pea seedlings (Pisum sativum L., Willet Wonder). Assays for calmodulin based on the activities of NAD kinase, bovine brain cyclic nucleotide phosphodiesterase, and human erythrocyte Ca/sup 2 -/-ATPase were compared for their sensitivities to calmodulin and for their abilities to discriminate between calmodulins from different sources. The activities of the three enzymes were determined in the presence of various concentrations of calmodulins from human erythrocyte, bovine brain, sea pansy (Renilla reniformis), mung bean seed (Vigna radiata L. Wilczek), mushroom (Agaricus bisporus), and Tetrahymena pyriformis. The concentrations of calmodulin required for 50% activation of the NAD kinase (K/sub 0.5/) ranged from 0.520 ng/ml for Tetrahymena to 2.20 ng/ml for bovine brain. The A/sub 0.5/ s ranged from 19.6 ng/ml for bovine brain calmodulin to 73.5 ng/ml for mushroom calmodulin for phosphodiesterase activation. The K/sub 0.5/'s for the activation of Ca/sup 2 +/-ATPase ranged from 36.3 ng/mol for erythrocyte calmodulin to 61.7 ng/ml for mushroom calmodulin. NAD kinase was not stimulated by phosphatidylcholine, phosphatidylserine, cardiolipin, or palmitoleic acid in the absence or presence of Ca/sup 2 +/. Palmitic acid had a slightly stimulatory effect in the presence of Ca/sup 2 +/ (10% of maximum), but no effect in the absence of Ca/sup 2 +/. Palmitoleic acid inhibited the calmodulin-stimulated activity by 50%. Both the NAD kinase assay and radioimmunoassay were able to detect calmodulin in extracts containing low concentrations of calmodulin. Estimates of calmodulin contents of crude homogenates determined by the NAD kinase assay were consistent with amounts obtained by various purification procedures. 30 references, 1 figure, 4 tables.

  14. Spectrophotometric validation of assay method for selected medicinal plant extracts


    Matthew Arhewoh; Augustine O. Okhamafe


    Objective: To develop UV spectrophotometric assay validation methods for some selected medicinal plant extracts.Methods: Dried, powdered leaves of Annona muricata (AM) and Andrographis paniculata (AP) as well as seeds of Garcinia kola (GK) and Hunteria umbellata (HU) were separately subjected to maceration using distilled water. Different concentrations of the extracts were scanned spectrophotometrically to obtain wavelengths of maximum absorbance. The different extracts were then subjected t...

  15. Purification of Intact Plant Protoplasts by Flotation at 1g

    Directory of Open Access Journals (Sweden)

    John Graham


    Full Text Available From a standard plant tissue digest adjusted to a density of 1.07 g/ml, protoplasts can be harvested by flotation through a low density barrier (1.03 g/ml. The delicate nature of these bodies is suited to this flotation strategy which can be carried out at 1g.

  16. Potential of using plant extracts for purification of shallow well water in Malawi (United States)

    Pritchard, M.; Mkandawire, T.; Edmondson, A.; O'Neill, J. G.; Kululanga, G.

    There has been very little scientific research work into the use of plant extracts to purify groundwater. Research studies on the purification of groundwater have mainly been carried out in developed countries and have focused on water purification systems using aluminium sulphate (a coagulant) and chlorine (a disinfectant). Such systems are expensive and not viable for rural communities due to abject poverty. Shallow well water, which is commonly available throughout Africa, is often grossly contaminated and usually consumed untreated. As a result, water-related diseases kill more than 5 million people every year worldwide. This research was aimed at examining natural plant extracts in order to develop inexpensive ways for rural communities to purify their groundwater. The study involved creating an inventory of plant extracts that have been used for water and wastewater purification. A prioritisation system was derived to select the most suitable extracts, which took into account criteria such as availability, purification potential, yield and cost of extraction. Laboratory trials were undertaken on the most promising plant extracts, namely: Moringa oleifera, Jatropha curcas and Guar gum. The extracts were added to water samples obtained from five shallow wells in Malawi. The trials consisted of jar tests to assess the coagulation potential and the resulting effect on physico-chemical and microbiological parameters such as temperature, pH, turbidity and coliforms. The results showed that the addition of M. oleifera, J. curcas and Guar gum can considerably improve the quality of shallow well water. Turbidity reduction was higher for more turbid water. A reduction efficiency exceeding 90% was achieved by all three extracts on shallow well water that had a turbidity of 49 NTU. A reduction in coliforms was about 80% for all extracts. The pH of the water samples increased with dosage, but remained within acceptable levels for drinking water for all the extracts

  17. Extraction and Purification of Depigmenting Agents from Chinese Plants

    Institute of Scientific and Technical Information of China (English)


    Depigmenting agents were solvent-extracted and purified by preparative and analytical HPLC from three Chinese plants; Chrysanthemum morifolium Ramat( Xizang Caijuhua), Rhodiola sachalinensis, and Terminalia chebula Retzius. Four fractions obtained from the ethyl ether layer of C. m. Rama and two fractions from the ethyl acetate layer of Rhodiola salientness show depigmenting effects. At δ 200, the ethyl acetate layers of Chrysanthemum morifolium Ramat, Rhodiola sachalinensis and the methanol extract of Terminalia chebula Retzius, can inhibit the melanin production of mouse B16 melanoma cells by 92%, 60% and 90%, respectively, whereas 46% inhibition was observed by commercially available depigmenting agents(arbutin). These results show the potential of these three Chinese plants as a novel resource for depigmenting agents in the cosmetic industry.

  18. Protocol: a rapid and economical procedure for purification of plasmid or plant DNA with diverse applications in plant biology

    Directory of Open Access Journals (Sweden)

    Li Li


    Full Text Available Abstract Research in plant molecular biology involves DNA purification on a daily basis. Although different commercial kits enable convenient extraction of high-quality DNA from E. coli cells, PCR and agarose gel samples as well as plant tissues, each kit is designed for a particular type of DNA extraction work, and the cost of purchasing these kits over a long run can be considerable. Furthermore, a simple method for the isolation of binary plasmid from Agrobacterium tumefaciens cells with satisfactory yield is lacking. Here we describe an easy protocol using homemade silicon dioxide matrix and seven simple solutions for DNA extraction from E. coli and A. tumefaciens cells, PCR and restriction digests, agarose gel slices, and plant tissues. Compared with the commercial kits, this protocol allows rapid DNA purification from diverse sources with comparable yield and purity at negligible cost. Following this protocol, we have demonstrated: (1 DNA fragments as small as a MYC-epitope tag coding sequence can be successfully recovered from an agarose gel slice; (2 Miniprep DNA from E. coli can be eluted with as little as 5 μl water, leading to high DNA concentrations (>1 μg/μl for efficient biolistic bombardment of Arabidopsis seedlings, polyethylene glycol (PEG-mediated Arabidopsis protoplast transfection and maize protoplast electroporation; (3 Binary plasmid DNA prepared from A. tumefaciens is suitable for verification by restriction analysis without the need for large scale propagation; (4 High-quality genomic DNA is readily isolated from several plant species including Arabidopsis, tobacco and maize. Thus, the silicon dioxide matrix-based DNA purification protocol offers an easy, efficient and economical way to extract DNA for various purposes in plant research.

  19. Protocol: a rapid and economical procedure for purification of plasmid or plant DNA with diverse applications in plant biology. (United States)

    Li, Jian-Feng; Li, Li; Sheen, Jen


    Research in plant molecular biology involves DNA purification on a daily basis. Although different commercial kits enable convenient extraction of high-quality DNA from E. coli cells, PCR and agarose gel samples as well as plant tissues, each kit is designed for a particular type of DNA extraction work, and the cost of purchasing these kits over a long run can be considerable. Furthermore, a simple method for the isolation of binary plasmid from Agrobacterium tumefaciens cells with satisfactory yield is lacking. Here we describe an easy protocol using homemade silicon dioxide matrix and seven simple solutions for DNA extraction from E. coli and A. tumefaciens cells, PCR and restriction digests, agarose gel slices, and plant tissues. Compared with the commercial kits, this protocol allows rapid DNA purification from diverse sources with comparable yield and purity at negligible cost. Following this protocol, we have demonstrated: (1) DNA fragments as small as a MYC-epitope tag coding sequence can be successfully recovered from an agarose gel slice; (2) Miniprep DNA from E. coli can be eluted with as little as 5 mul water, leading to high DNA concentrations (>1 mug/mul) for efficient biolistic bombardment of Arabidopsis seedlings, polyethylene glycol (PEG)-mediated Arabidopsis protoplast transfection and maize protoplast electroporation; (3) Binary plasmid DNA prepared from A. tumefaciens is suitable for verification by restriction analysis without the need for large scale propagation; (4) High-quality genomic DNA is readily isolated from several plant species including Arabidopsis, tobacco and maize. Thus, the silicon dioxide matrix-based DNA purification protocol offers an easy, efficient and economical way to extract DNA for various purposes in plant research.

  20. Gas purification downstream of waste incineration plants. Gasreinigung hinter Muellverbrennungsanlagen

    Energy Technology Data Exchange (ETDEWEB)

    Hoelter, H.; Igelbuescher, H.; Gresch, H.; Dewert, H.


    HCl, HF and also SO/sub 2/ as well as most heavy metals can be separated in dry filters of refuse incineration plants by adding lime hydrate, but the gaseous heavy metals, such as mercury, can only be removed in a secondary wet separator. A secondary wet separator is installed for the separation of the residual acid emission components which are still in the flue gas after passing through the dry filter. Calcium hydroxide solutions and, additionally, an active substance of trimercapto-s-triazine are added to the scrubbing water cycle of this separator.

  1. A simple in vitro acylation assay based on optimized HlyA and HlyC purification. (United States)

    Thomas, Sabrina; Smits, Sander H J; Schmitt, Lutz


    HlyA is a toxin secreted by uropathogenic Escherichia coli strains. HlyA belongs to the repeats in the toxin protein family and needs (i) a posttranslational, fatty acylation at two internal lysines by the acyltransferase HlyC and (ii) extracellular ion binding to achieve its active conformation. Both processes are not fully understood and experiments are often limited due to the low amounts of protein available. Here, we present an optimized purification protocol for the proteins involved in HlyA activation as well as a quick and nonradioactive assay for in vitro HlyA acylation. These may simplify future experiments, e.g., activity scanning and characterization of HlyA or HlyC mutants as demonstrated with single and double HlyA lysine mutants. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. Spectrophotometric validation of assay method for selected medicinal plant extracts

    Directory of Open Access Journals (Sweden)

    Matthew Arhewoh


    Full Text Available Objective: To develop UV spectrophotometric assay validation methods for some selected medicinal plant extracts.Methods: Dried, powdered leaves of Annona muricata (AM and Andrographis paniculata (AP as well as seeds of Garcinia kola (GK and Hunteria umbellata (HU were separately subjected to maceration using distilled water. Different concentrations of the extracts were scanned spectrophotometrically to obtain wavelengths of maximum absorbance. The different extracts were then subjected to validation studies following international guidelines at the respective wavelengths obtained.Results: The results showed linearity at peak wavelengths of maximum absorbance of 292, 280, 274 and 230 nm for GK, HU, AM and AP, respectively. The calibration curves for the different concentrations of the extract gave R2 values ranging from 0.9831 for AM to 0.9996 for AP the inter-day and intra-day precision study showed that the relative standard deviation (% was ≤ 10% for all the extracts.Conclusion: The aqueous extracts and isolates of these plants can be assayed and monitored using these wavelengths.

  3. A cost-effective ELP-intein coupling system for recombinant protein purification from plant production platform.

    Directory of Open Access Journals (Sweden)

    Li Tian

    Full Text Available BACKGROUND: Plant bioreactor offers an efficient and economical system for large-scale production of recombinant proteins. However, high cost and difficulty in scaling-up of downstream purification of the target protein, particularly the common involvement of affinity chromatography and protease in the purification process, has hampered its industrial scale application, therefore a cost-effective and easily scale-up purification method is highly desirable for further development of plant bioreactor. METHODOLOGY/PRINCIPAL FINDINGS: To tackle this problem, we investigated the ELP-intein coupling system for purification of recombinant proteins expressed in transgenic plants using a plant lectin (PAL with anti-tumor bioactivity as example target protein and rice seeds as production platform. Results showed that ELP-intein-PAL (EiP fusion protein formed novel irregular ER-derived protein bodies in endosperm cells by retention of endogenous prolamins. The fusion protein was partially self-cleaved in vivo, but only self-cleaved PAL protein was detected in total seed protein sample and deposited in protein storage vacuoles (PSV. The in vivo uncleaved EiP protein was accumulated up to 2-4.2% of the total seed protein. The target PAL protein could be purified by the ELP-intein system efficiently without using complicated instruments and expensive chemicals, and the yield of pure PAL protein by the current method was up to 1.1 mg/g total seed protein. CONCLUSION/SIGNIFICANCE: This study successfully demonstrated the purification of an example recombinant protein from rice seeds by the ELP-intein system. The whole purification procedure can be easily scaled up for industrial production, providing the first evidence on applying the ELP-intein coupling system to achieve cost-effective purification of recombinant proteins expressed in plant bioreactors and its possible application in industry.

  4. Purification of Active Myrosinase from Plants by Aqueous Two-Phase Counter-Current Chromatography (United States)

    Wade, Kristina L.; Ito, Yoichiro; Ramarathnam, Aarthi; Holtzclaw, W. David; Fahey, Jed W.


    Introduction Myrosinase (thioglucoside glucohydrolase; E.C., is a plant enzyme of increasing interest and importance to the biomedical community. Myrosinase catalyses the formation of isothiocyanates such as sulforaphane (frombroccoli) and 4-(α-l-rhamnopyranosyloxy)benzyl isothiocyanate (from moringa), which are potent inducers of the cytoprotective phase-2 response in humans, by hydrolysis of their abundant glucosinolate (β-thioglucoside N-hydroxysulphate) precursors. Objective To develop an aqueous two-phase counter-current chromatography (CCC) system for the rapid, three-step purification of catalytically active myrosinase. Methods A high-concentration potassium phosphate and polyethylene glycol biphasic aqueous two-phase system (ATPS) is used with a newly developed CCC configuration that utilises spiral-wound, flat-twisted tubing (with an ovoid cross-section). Results Making the initial crude plant extract directly in the ATPS and injecting only the lower phase permitted highly selective partitioning of the myrosinase complex before a short chromatography on a spiral disk CCC. Optimum phase retention and separation of myrosinase from other plant proteins afforded a 60-fold purification. Conclusion Catalytically active myrosinase is purified from 3-day broccoli sprouts, 7-day daikon sprouts, mustard seeds and the leaves of field-grown moringa trees, in a CCC system that is predictably scalable. PMID:25130502

  5. A study of naturally occurring radon in Swedish water purification plants.



    Radon dissolved in drinking-water can be transferred into the indoor air and is one of the main transfer pathways for radon. At water purification plants, large quantities of water are treated and there is a risk that radon degasses from the water and enters into the indoor air. Hence, there is a risk for elevated radon levels in the indoor air at these facilities. This study aims to investigate the general impact of water treatment processes on the radon concentration in water and its transf...

  6. Air purification in industrial plants producing automotive rubber components in terms of energy efficiency (United States)

    Grzebielec, Andrzej; Rusowicz, Artur; Szelągowski, Adam


    In automotive industry plants, which use injection molding machines for rubber processing, tar contaminates air to such an extent that air fails to enter standard heat recovery systems. Accumulated tar clogs ventilation heat recovery exchangers in just a few days. In the plant in which the research was conducted, tar contamination causes blockage of ventilation ducts. The effect of this phenomenon was that every half year channels had to be replaced with new ones, since the economic analysis has shown that cleaning them is not cost-efficient. Air temperature inside such plants is often, even in winter, higher than 30°C. The air, without any means of heat recovery, is discharged outside the buildings. The analyzed plant uses three types of media for production: hot water, cold water at 14°C (produced in a water chiller), and compressed air, generated in a unit with a rated power consumption of 180 kW. The aim of the study is to determine the energy efficiency improvement of this type of manufacturing plant. The main problem to solve is to provide an air purification process so that air can be used in heat recovery devices. The next problem to solve is to recover heat at such a temperature level that it would be possible to produce cold for technological purposes without air purification. Experimental studies have shown that air purification is feasible. By using one microjet head, a total of 75% of tar particles was removed from the air; by using 4 heads, a purification efficiency of 93% was obtained. This method of air purification causes air temperature to decrease from 35°C to 20°C, which significantly reduces the potential for heat recovery. The next step of the research was designing a cassette-plate heat exchanger to exchange heat without air purification. The economic analysis of such a solution revealed that replacing the heat exchanger with a new one even once a year was not cost-efficient. Another issue examined in the context of energy efficiency was

  7. Air purification in industrial plants producing automotive rubber components in terms of energy efficiency

    Directory of Open Access Journals (Sweden)

    Grzebielec Andrzej


    Full Text Available In automotive industry plants, which use injection molding machines for rubber processing, tar contaminates air to such an extent that air fails to enter standard heat recovery systems. Accumulated tar clogs ventilation heat recovery exchangers in just a few days. In the plant in which the research was conducted, tar contamination causes blockage of ventilation ducts. The effect of this phenomenon was that every half year channels had to be replaced with new ones, since the economic analysis has shown that cleaning them is not cost-efficient. Air temperature inside such plants is often, even in winter, higher than 30°C. The air, without any means of heat recovery, is discharged outside the buildings. The analyzed plant uses three types of media for production: hot water, cold water at 14°C (produced in a water chiller, and compressed air, generated in a unit with a rated power consumption of 180 kW. The aim of the study is to determine the energy efficiency improvement of this type of manufacturing plant. The main problem to solve is to provide an air purification process so that air can be used in heat recovery devices. The next problem to solve is to recover heat at such a temperature level that it would be possible to produce cold for technological purposes without air purification. Experimental studies have shown that air purification is feasible. By using one microjet head, a total of 75% of tar particles was removed from the air; by using 4 heads, a purification efficiency of 93% was obtained. This method of air purification causes air temperature to decrease from 35°C to 20°C, which significantly reduces the potential for heat recovery. The next step of the research was designing a cassette-plate heat exchanger to exchange heat without air purification. The economic analysis of such a solution revealed that replacing the heat exchanger with a new one even once a year was not cost-efficient. Another issue examined in the context of


    Directory of Open Access Journals (Sweden)

    Carmen Alice Teacă


    Full Text Available Environmental pollution has a harmful action on bioresources, including agricultural crops. It is generated through many industrial activities such as mining, coal burning, chemical technology, cement production, pulp and paper industry, etc. The toxicity of different industrial wastes and heavy metals excess was evaluated using crop plant assays (germination and hydroponics seedlings growth tests. Experimental data regarding the germination process of wheat (from two cultivars and rye seeds in the presence of industrial wastes (thermal power station ash, effluents from a pre-bleaching stage performed on a Kraft cellulose – chlorinated lignin products or chlorolignin, along with use of an excess of some heavy metals (Zn and Cu are presented here. Relative seed germination, relative root elongation, and germination index (a factor of relative seed germination and relative root elongation were determined. Relative root elongation and germination index were more sensitive indicators of toxicity than seed germination. The toxic effects were also evaluated in hydroponics experiments, the sensitivity of three crop plant species, namely Triticum aestivum L. (wheat, Secale cereale (rye, and Zea mays (corn being compared. Physiological aspects, evidenced both by visual observation and biometric measurements (mean root, aerial part and plant length, as well as the cellulose and lignin content were examined.

  9. Symbiotic properties of Bradyrhizobium sp. (Lupinus assayed on serradella plants

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    Mieczysława Deryło


    Full Text Available Physiological and symbiotic properties of Bradyrhizobium sp. (Lupinus nodule isolates were compared to the standard slow-growing Bradyrhizobium sp. (Lupinus strain USDA 3045. Lupine nodules isolates showed typical characteristics for bradyrhizobial strains and nodulated small seed legume, serradella (Ornithopus sativus, in tube test. We observed a permanent physiological segregation of the effective (Fix' and ineffective (Fix- symbiotic phenotype for all tested bradyrhizobial strains during the growth of serradella in plant tube test. The ultrastructural differences between Fix* and Fix serradella nodules were observed. Rapid and visible nodulation as well as easy assay of the reduction of acetylene make serradella a convenient system for studies of Bradyrhizobium sp. (Lupinus strains in laboratory conditions.

  10. [Selection of winter plant species for wetlands constructed as sewage treatment systems and evaluation of their wastewater purification potentials]. (United States)

    Chen, Yong-hua; Wu, Xiao-fu; Chen, Ming-li; Jiang, Li-juan; Li, Ke-lin; Lei, Dian; Wang, Hai-bin


    In order to establish an evaluation system for selection of winter wetland plants possessing high wastewater purification potentials in subtropics areas, designed sewage treatment experiments were carried out by introducing into the constructed wetlands 25 species of winter wetland plants. Cluster analysis was performed by including harmful environment-resistant enzyme and substrate enzyme activities into the commonly applied plant screening and assessment indexes system. The obtained results indicated that there were significant differences among the tested winter plants in their root length and vigor, leaf malonaldehyde (MDA), biomass, average nitrogen and phosphorus concentration and uptake, and urease and phosphoric acid enzyme activities in the root areas. Based on the established evaluation system, the tested plants were clustered into 3 groups. The plants in the 1st group possessing high purification potentials are Oenanthe javanica, Brassicacapestris, Juncus effusu, Saxifragaceae, Iris pseudoacorus, Osmanthus fragrans and Iris ensata; those in the 2nd group possessing moderate purification potentials are Brassica oleracea var acephala, Calendula officinalis, Aucuba japonica, Ligustrum lucidu, Beta vulgaris, Rhododendron simsii and Ilex latifolia; and those in the 3rd group with low purification potentials are Brassica oleracea var acephala, Calistephus chinensis, Rosa chinensis, Antirrhinums, Liriope palatyphylla, Zephyranthes candida, Fatshedera lizei, Petunia hybrida, Ilex quihoui, Dianthus caryophyllus and Loropetalum chinensis.

  11. [Selection and purification potential evaluation of woody plant in vertical flow constructed wetlands in the subtropical area]. (United States)

    Chen, Yong-Hua; Wu, Xiao-Fu; Hao, Jun; Chen, Ming-Li; Zhu, Guang-Yu


    In order to solve the problem that wetland herbaceous plants tend to die during winter in subtropics areas, selection and purification potential evaluation experiments were carried out by introducing into the constructed wetlands 16 species of woody wetland plants. Cluster analysis was performed by including the morphological characteristics, physiological characteristics, as well as nitrogen and phosphorus accumulation of the woody wetland plants. The results indicated that there were significant differences among the tested woody plants in their survival rate, height increase, root length increase and vigor, Chlorophyll content, Superoxide dismutase, Malonaldehyde, Proline, Peroxidase, biomass, average concentration and accumulation of nitrogen and phosphorus. Based on the established evaluation system, the tested plants were clustered into 3 groups. The plants in the 1st group possessing high purification potentials are Nerium oleander and Hibiscus syriacus. Those in the 2nd group possessing moderate purification potentials are Trachycarpus fortune, Llex latifolia Thunb., Gardenia jasminoides, Serissa foetida and Ilex crenatacv Convexa. And those in the 3rd group with low purification potentials are Jasminum udiflorum, Hedera helix, Ligustrum vicaryi, Ligustrum lucidum, Buxus sempervives, Murraya paniculata, Osmanthus fragrans, Mahoniafortune and Photinia serrulata.

  12. High yield purification of Plasmodium falciparum merozoites for use in opsonizing antibody assays. (United States)

    Hill, Danika L; Eriksson, Emily M; Schofield, Louis


    Plasmodium falciparum merozoite antigens are under development as potential malaria vaccines. One aspect of immunity against malaria is the removal of free merozoites from the blood by phagocytic cells. However assessing the functional efficacy of merozoite specific opsonizing antibodies is challenging due to the short half-life of merozoites and the variability of primary phagocytic cells. Described in detail herein is a method for generating viable merozoites using the E64 protease inhibitor, and an assay of merozoite opsonin-dependent phagocytosis using the pro-monocytic cell line THP-1. E64 prevents schizont rupture while allowing the development of merozoites which are released by filtration of treated schizonts.  Ethidium bromide labelled merozoites are opsonized with human plasma samples and added to THP-1 cells. Phagocytosis is assessed by a standardized high throughput protocol. Viable merozoites are a valuable resource for assessing numerous aspects of P. falciparum biology, including assessment of immune function. Antibody levels measured by this assay are associated with clinical immunity to malaria in naturally exposed individuals. The assay may also be of use for assessing vaccine induced antibodies.  

  13. Occurrence of selected pharmaceuticals at drinking water purification plants in Japan and implications for human health. (United States)

    Simazaki, Dai; Kubota, Reiji; Suzuki, Toshinari; Akiba, Michihiro; Nishimura, Tetsuji; Kunikane, Shoichi


    The present study was performed to determine the occurrence of 64 pharmaceuticals and metabolites in source water and finished water at 6 drinking water purification plants and 2 industrial water purification plants across Japan. The analytical methods employed were sample concentration using solid-phase extraction cartridges and instrumental analysis by liquid chromatography with tandem mass spectrometry (LC-MS/MS), liquid chromatography with mass spectrometry (LC/MS), or trimethylsilyl derivatization followed by gas chromatography with mass spectrometry (GC/MS). Thirty-seven of the 64 target substances were detected in the source water samples. The maximum concentrations in the source water were mostly below 50 ng/L except for 13 substances. In particular, residual concentrations of iopamidol (contrast agent) exceeded 1000 ng/L at most facilities. Most of the residual pharmaceuticals and metabolites in the source water samples were removed in the course of conventional and/or advanced drinking water treatments, except for 7 pharmaceuticals and 1 metabolite, i.e., amantadine, carbamazepine, diclofenac, epinastine, fenofibrate, ibuprofen, iopamidol, and oseltamivir acid. The removal ratios of the advanced water treatment processes including ozonation and granular activated carbon filtration were typically much higher than those of the conventional treatment processes. The margins of exposure estimated by the ratio of daily minimum therapeutic dose to daily intake via drinking water were substantial, and therefore the pharmacological and physiological impacts of ingesting those residual substances via drinking water would be negligible. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Centrifugal gas-phase transition magnetophoresis (GTM)--a generic method for automation of magnetic bead based assays on the centrifugal microfluidic platform and application to DNA purification. (United States)

    Strohmeier, Oliver; Emperle, Alexander; Roth, Günter; Mark, Daniel; Zengerle, Roland; von Stetten, Felix


    Transportation of magnetic beads between different reagents plays a crucial role in many biological assays e.g. for purification of biomolecules or cells where the beads act as a mobile solid support. Therefore, usually a complex set-up either for fluidic processing or for manipulation of magnetic beads is required. To circumvent these drawbacks, we present a facile and automated method for the transportation of magnetic beads between multiple microfluidic chambers on a centrifugal microfluidic cartridge "LabDisk". The method excels by requiring only one stack of stationary permanent magnets, a specific microfluidic layout without actively controlled valves and a predefined frequency protocol for rotation of the LabDisk. Magnetic beads were transported through three fluidically separated chambers with a yield of 82.6% ± 3.6%. Bead based DNA purification from a dilution series of a Listeria innocua lysate and from a lambda phage DNA standard was demonstrated where the three chambers were used for binding, washing and elution of DNA. Recovery of L. innocua DNA was up to 68% ± 24% and for lambda phage DNA 43% ± 10% compared to manual reference purification in test tubes. Complete purification was conducted automatically within 12.5 min. Since all reagents can be preloaded onto the LabDisk prior to purification, no further hands-on steps are required during processing. Due to its modular and generic character, the presented method could also be adapted to other magnetic bead based assays e.g. to immunoassays or protein affinity purification, solely requiring the adjustment of number and volumes of the fluidic chambers.

  15. Purification of Protein Chaperones and Their Functional Assays with Intermediate Filaments. (United States)

    Perng, Ming-Der; Huang, Yu-Shan; Quinlan, Roy A


    Intermediate filament (IF) scaffolds facilitate small heat shock protein (sHSP) function, while IF function is sHSP dependent. sHSPs interact with IFs and the importance of this interaction is to maintain the individuality of the IFs and to modulate interfilament interactions both in networks and in assembly intermediates. Mutations in both sHSPs and their interacting IF proteins phenocopy each other in the human diseases they cause. This establishes a key functional relationship between these two very distinct protein families, and it also evidences the role of this cytoskeleton-chaperone complex in the cellular stress response. In this chapter, we describe the detailed experimental protocols for the preparation of purified IF proteins and sHSPs to facilitate the study in vitro of their functional interactions. In addition, we describe the detailed biochemical procedures to assess the effect of sHSP on the assembly of IFs, the binding to IFs, and the prevention of noncovalent filament-filament interactions using in vitro cosedimentation, electron microscopy, and viscosity assays. These assays are valuable research tools to study and manipulate sHSP-IF complexes in vitro and therefore to determine the structure-function detail of this complex, and how it contributes to cellular, tissue, and organismal homeostasis and the in vivo stress response.

  16. Expression, Purification and Activity Assay of the Recombinant Protein of Catechol-O-Methyltransferase from Chinese White Shrimp (Fenneropenaeus chinensis

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    Dian-Xiang Li


    Full Text Available Problem statement: We have previously cloned a gene of Chinese white shrimp Catechol O-Methyltransferase (designated Fc-COMT and characterized the gene expression pattern. In this study, expression and purification as well as activity assay of the recombinant Fc-COMT was further conducted. Approach: Using pET-30a (+ as a prokaryotic expression vector, the recombinant Fc- COMT was expressed in the supernatant of Escherichia coli lysate and easily purified by His-Bind resin chromatography. SDS-PAGE analysis showed that the molecular mass of recombinant Fc-COMT was approximately 30,000 Da, in good agreement with the software-predicted molecular weight. The enzymatic activity of recombinant Fc-COMT was tested using Dihydroxybenzoic Acid (DHBAc as a substrate. Results: The methyl products of DHBAc, Vanillic Acid (VA and Isovanillic Acid (IVA, were detected in the enzymatic reaction mixture with recombinant Fc-COMT by High Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS. Conclusion: The recombinant Fc-COMT has catalytic activity of transferring methyl group from S-Adenosyl-L-Methionine (SAM to the 3’ hydroxyl or 4’ hydroxyl group of benzyl ring of DHBAc.

  17. Use of biotin-labeled nucleic acids for protein purification and agarose-based chemiluminescent electromobility shift assays. (United States)

    Rodgers, J T; Patel, P; Hennes, J L; Bolognia, S L; Mascotti, D P


    We have employed biotin-labeled RNA to serve two functions. In one, the biotin tethers the RNA to streptavidin-agarose beads, creating an affinity resin for protein purification. In the other, the biotin functions as a label for use in a modified chemiluminescent electromobility shift assay (EMSA), a technique used to detect the formation of protein-RNA complexes. The EMSA that we describe avoids the use not only of radioactivity but also of neurotoxic acrylamide by using agarose as the gel matrix in which the free nucleic acid is separated from protein-nucleic acid complexes. After separation of free from complexed RNA in agarose, the RNA is electroblotted to positively charged nylon. The biotin-labeled RNA is readily bound by a streptavidin-alkaline phosphatase conjugate, allowing for very sensitive chemiluminescent detection ( approximately 0.1-1.0 fmol limit). Using our system, we were able to purify both known iron-responsive proteins (IRPs) from rat liver and assess their binding affinity to RNA containing the iron-responsive element (IRE) using the same batch of biotinylated RNA. We show data indicating that agarose is especially useful for cases when large complexes are formed, although smaller complexes are even better resolved.

  18. Protein body-inducing fusions for high-level production and purification of recombinant proteins in plants. (United States)

    Conley, Andrew J; Joensuu, Jussi J; Richman, Alex; Menassa, Rima


    For the past two decades, therapeutic and industrially important proteins have been expressed in plants with varying levels of success. The two major challenges hindering the economical production of plant-made recombinant proteins include inadequate accumulation levels and the lack of efficient purification methods. To address these limitations, several fusion protein strategies have been recently developed to significantly enhance the production yield of plant-made recombinant proteins, while simultaneously assisting in their subsequent purification. Elastin-like polypeptides are thermally responsive biopolymers composed of a repeating pentapeptide 'VPGXG' sequence that are valuable for the purification of recombinant proteins. Hydrophobins are small fungal proteins capable of altering the hydrophobicity of their respective fusion partner, thus enabling efficient purification by surfactant-based aqueous two-phase systems. Zera, a domain of the maize seed storage protein γ-zein, can induce the formation of protein storage bodies, thus facilitating the recovery of fused proteins using density-based separation methods. These three novel protein fusion systems have also been shown to enhance the accumulation of a range of different recombinant proteins, while concurrently inducing the formation of protein bodies. The packing of these fusion proteins into protein bodies may exclude the recombinant protein from normal physiological turnover. Furthermore, these systems allow for quick, simple and inexpensive nonchromatographic purification of the recombinant protein, which can be scaled up to industrial levels of protein production. This review will focus on the similarities and differences of these artificial storage organelles, their biogenesis and their implication for the production of recombinant proteins in plants and their subsequent purification.

  19. Expression, Purification and Activity Assay of Two New Recombinant Antagonists of Fibrinogen Receptor

    Directory of Open Access Journals (Sweden)

    Jianbo Yang


    Full Text Available The gene sequence of Decorsin which is extracted from a kind of North American leeches was synthesized. Two recombinant proteins, Annexin V plus Decorsin (AnnV-D39 and Annexin V plus the carboxyl terminal 27 amino acid residues of Decorsin(AnnV-D27, were constructed. And a 10 amino acids linker peptide of GGGGSGGGGS was inserted between Annexin V and Decorsin in AnnV-D39. Using pET-28(a+ as an expressing vector, both two recombinant proteins were expressed in E. Coli BL21(DE3 with high efficiency as inclusion bodies. The expression products were purified by DEAE-Cellulose 52 and Sepharose CL-4B chromatography under denaturing condition. Platelet Aggregation Assay (PAA shows that AnnV-D39 has good anti-platelet aggregation activity. However, AnnV-D27 shows no such activities in any PAA test. AnnV-D39 shows good anti-platelet aggregation activity as a new antagonist of fibrinogen receptor, while Annv-D27 needs re-modification

  20. Phytotechnological purification of water and bio energy utilization of plant biomass (United States)

    Stom, D. I.; Gruznych, O. V.; Zhdanova, G. O.; Timofeeva, S. S.; Kashevsky, A. V.; Saksonov, M. N.; Balayan, A. E.


    The aim of the study was to explore the possibility of using the phytomass of aquatic plants as the substrate in the microbial fuel cells and selection of microorganisms suitable for the generation of electricity on this substrate. The conversion of chemical energy of phytomass of aquatic plants to the electrical energy was carried out in a microbial fuel cells by biochemical transformation. As biological agents in the generation of electricity in the microbial fuel cells was used commercial microbial drugs “Doctor Robic 109K” and “Vostok-EM-1”. The results of evaluation of the characteristics of electrogenic (amperage, voltage) and the dynamics of the growth of microorganisms in the microbial fuel cells presents in the experimental part. As a source of electrogenic microorganisms is possible to use drugs “Dr. Robic 109K” and “Vostok-EM-1” was established. The possibility of utilization of excess phytomass of aquatic plants, formed during the implementation of phytotechnological purification of water, in microbial fuel cells, was demonstrated. The principal possibility of creating hybrid phytotechnology (plant-microbe cells), allowing to obtain electricity as a product, which can be used to ensure the operation of the pump equipment and the creation of a full cycle of resource-saving technologies for water treatment, was reviewed.

  1. Generic chromatography-based purification strategies accelerate the development of downstream processes for biopharmaceutical proteins produced in plants. (United States)

    Buyel, Johannes F; Fischer, Rainer


    Plants offer a valuable alternative to cultured mammalian cells for the production of recombinant biopharmaceutical proteins. However, the target protein typically represents only a minor fraction of the total protein in the initial plant extract, which means that the development of product-specific chromatography-based purification strategies is often laborious and expensive. To address this challenge, we designed a generic downstream process that is suitable for the purification of recombinant proteins with diverse properties from plant production platforms. This was achieved by focusing on the binding behavior of tobacco host cell proteins (HCPs) to a broad set of chromatography resins under different pH and conductivity conditions. Strong cation exchanger and salt-tolerant anion exchanger resins exhibited the best resolution of tobacco HCPs among the 13 tested resins, and their selectivity was easy to manipulate through the adjustment of pH and conductivity. The advantages, such as direct capture of a target protein from leaf extract, and limitations, such as low binding capacity, of various chromatography ligands and resins are discussed. We also address the most useful applications of the chromatography ligands, namely recovery of proteins with a certain pI, in a downstream process that aims to purify diverse plant-derived biopharmaceutical proteins. Based on these results, we describe generic purification schemes that are suitable for acidic, neutral, and basic target proteins, as a first step toward the development of industrial platform processes.

  2. [Methods of hygromycin B phosphotransferase activity assay in transgenic plant]. (United States)

    Zhuo, Qin; Yang, Xiaoguang


    Hygromycin B phosphotransferase (HPT) is a widely used selectable marker protein of transgenic plant. Detection of its activity is critical to studies on the development of various transgenic plants, silence of inserted gene, marker-free system development and safety assessment of transgenic food. In this paper, several methods for detecting the activity of this enzyme were reviewed.

  3. In vitro CLE peptide bioactivity assay on plant roots (United States)

    Plant CLAVATA3/ESR (CLE)-related proteins play diverse roles in plant growth and development including regulating the development of root meristem. Mature CLE peptides are typically 12-13 amino acids (aa) in length that are derived from the conserved C-termini of their precursor proteins. Genes enco...

  4. Development of an automated scoring system for plant comet assay

    Directory of Open Access Journals (Sweden)

    Bertrand Pourrut


    -\tnucleus density: increase the density of nuclei is of importance to increase scoring reliability (Sharma et al., 2012. In conclusion, increasing plant nucleus extraction yield and automated scoring of nuclei do represent big challenges. However, our promising preliminary results open up the perspective of an automated high-throughput scoring of plant nuclei.

  5. Functional heterologous expression and purification of a mammalian methyl-CpG binding domain in suitable yield for DNA methylation profiling assays. (United States)

    Boyd, Mary E; Heimer, Brandon W; Sikes, Hadley D


    DNA methylation is a major epigenetic modification in mammalian cells, and patterns involving methylation of cytosine bases, known as CpG methylation, have been implicated in the development of many types of cancer. Methyl binding domains (MBDs) excised from larger mammalian methyl-CpG-binding proteins specifically recognize methyl-cytosine bases of CpG dinucleotides in duplex DNA. Previous molecular diagnostic studies involving MBDs have employed Escherichia coli for protein expression with either low soluble yields or the use of time-consuming denaturation-renaturation purification procedures to improve yields. Efficient MBD-based diagnostics require expression and purification methods that maximize protein yield and minimize time and resource expenditure. This study is a systematic optimization analysis of MBD expression using both SDS-PAGE and microscopy and it provides a comparison of protein yield from published procedures to that from the conditions found to be optimal in these experiments. Protein binding activity and specificity were verified using a DNA electrophoretic mobility shift assay, and final protein yield was improved from the starting conditions by a factor of 65 with a simple, single-step purification.

  6. Purification, characterization and stability of barley grain peroxidase BP1, a new type of plant peroxidase

    DEFF Research Database (Denmark)

    Rasmussen, Christine B; Henriksen, Anette; Abelskov, A. Katrine


    The major peroxidase of barley grain (BP 1) has enzymatic and spectroscopic properties that are very differeant from those of other known plant peroxidases (EC and can therefore contribute to the understanding of the many physiological functions ascribed to these enzymes. To study...... the structure-function relationships of this unique model peroxidase, large-scale and laboratory-scale purifications have been developed. The two batches of pure BP 1 obtained were identical in their enzymatic and spectral properties, and confirmed that BP 1 is different from the prototypical horseradish...... peroxidase isoenzyme C (HRP C). However, when measuring the specific activity of BP 1 at pH 4.0 in the presence of 1 mM CaCl2, the enzyme was as competent as HRP C at neutral pH towards a variety of substrates (mM mg(-1) min(-1)): coniferyl alcohol (930+/-48), caffeic acid (795+/-53), ABTS (2,2(1)-azino...

  7. Ammonia nitrogen removal from acetylene purification wastewater from a PVC plant by struvite precipitation. (United States)

    Zhu, Lei; Dong, DeMing; Hua, XiuYi; Guo, ZhiYong; Liang, DaPeng

    Acetylene purification wastewater (APW) usually contains high concentrations of ammonia nitrogen (NH4-N), which is generated during the production of acetylene in a polyvinylchloride manufacturing plant. In this study, a struvite precipitation method was selected to remove NH4-N from the APW. Laboratory-scale batch experiments were performed to investigate the effects of the initial APW pH, phosphate (PO4(3-)) concentration, magnesium (Mg(2+)) concentration, and sources of PO4(3-) and Mg(2+) on NH4-N removal. The results indicated that the initial APW pH had a significant effect on the removal of NH4-N, while the other factors had relatively minor effect. The NH4-N could be effectively removed at an optimum initial APW pH of 9.5, when Na2HPO4·12H2O and MgSO4·7H2O were both applied to NH4-N at a ratio of 1.2. Under these conditions, the efficiency of removal of NH4-N, total nitrogen and chemical oxygen demand were 85%, 84% and 18%, respectively. The X-ray diffraction analysis indicated that the precipitates were dominated by struvite. The scanning electron microscopy analysis of the precipitates showed a typical morphology of stick-like and prismatic crystals with coarse surface. The energy dispersive spectroscopy analysis indicated that the precipitates contained P, O, Mg and Ca.

  8. Optimization of a homogeneous assay for kinase inhibitors in plant extracts. (United States)

    Dufau, Isabelle; Lazzari, Anne; Samson, Arnaud; Pouny, Isabelle; Ausseil, Frederic


    To identify natural and original kinase inhibitors from plant extracts, we have developed and compared a heterogeneous enzyme-linked immunosorbent assay (ELISA) and a homogeneous time-resolved fluorescence (HTRF, Cisbio International, Bagnols/Cèze, France) assay. Kinase affinity for the ATP substrate was determined in both assays, and the same [ATP]/ATP Km ratio was used in each case to enable the identification of ATP competitive and noncompetitive inhibitors. Assays were then used to screen the same collection of chemical compounds and plant extracts. The intra-assay correlation analysis of each technology showed a very good screening precision in HTRF and an acceptable one in ELISA. When the two methods were compared, a poor correlation was obtained with a higher hit rate in the ELISA. We then performed a detailed study of the ELISA hits and showed that they also presented a strong antioxidant activity, associated with high adsorption into microplate wells, which interfered with the horseradish peroxidase-based detection system. These hits were then flagged as false-positives. We also showed that many plant extracts presented this kind of activity and that this interference could explain the lack of correlation between the assays. These findings suggest that assay design should be carefully adapted to the substances to be screened and that interferences should be extensively considered before any assay development process and comparison studies. In spite of a few interferences, our results showed that a homogeneous-phase assay like the HTRF assay could be more efficiently used for plant extract screening than a heterogeneous-phase assay like ELISA.

  9. Purification and assays of Rhodobacter capsulatus RegB-RegA two-component signal transduction system. (United States)

    Swem, Lee R; Swem, Danielle L; Wu, Jiang; Bauer, Carl E


    Two-component signal-transduction systems, composed of a histidine-sensor kinase and a DNA-binding response regulator, allow bacteria to detect environmental changes and adjust cellular physiology to live more efficiently in a broad distribution of niches. Although many two-component signal-transduction systems are known, a limited number of signals that stimulate these systems have been discovered. This chapter describes the purification and characterization of the predominant two-component signal-transduction system utilized by Rhodobacter capsulatus, a nonsulfur purple photosynthetic bacterium. Specifically, we explain the overexpression, detergent solubilization, and purification of the full-length membrane-spanning histidine-sensor kinase RegB. We also provide a method to measure autophosphorylation of RegB and discern the effect of its signal molecule, ubiquinone, on autophosphorylation levels. In addition we describe the overexpression and purification of the cognate response regulator RegA and a technique used to visualize the phosphotransfer reaction from RegB to RegA.

  10. An assay for secologanin in plant tissues based on enzymatic conversion into strictosidine

    DEFF Research Database (Denmark)

    Hallard, Didier; van der Heijden, Robert; Contin, Adriana


    The secoiridoid glucoside secologanin is the terpenoid building block in the biosynthesis of terpenoid indole alkaloids. A method for its determination in plant tissues and cell suspension cultures has been developed. This assay is based on the condensation of secologanin with tryptamine, yielding...... of STR for secologanin, in combination with a sensitive and selective HPLC system, allows a simple extraction of secologanin from plant tissue. The detection limit of this methos is 15 ng secologanin. Using this assay, secologanin contents were determined in tissues of various plant species; Lonicera...... xylosteum hairy roots were found to contain 1 % of secologanin on a dry weight basis....

  11. Development of Rapid Isothermal Amplification Assays for Detection of Phytophthora spp. in Plant Tissue. (United States)

    Miles, Timothy D; Martin, Frank N; Coffey, Michael D


    Several isothermal amplification techniques recently have been developed that are tolerant of inhibitors present in many plant extracts, which can reduce the need for obtaining purified DNA for running diagnostic assays. One such commercially available technique that has similarities with real-time polymerase chain reaction (PCR) for designing primers and a labeled probe is recombinase polymerase amplification (RPA). This technology was used to develop two simple and rapid approaches for detection of Phytophthora spp.: one genus-specific assay multiplexed with a plant internal control and the other species-specific assays for Phytophthora ramorum and P. kernoviae. All assays were tested for sensitivity (ranging from 3 ng to 1 fg of DNA) and specificity using DNA extracted from more than 136 Phytophthora taxa, 21 Pythium spp., 1 Phytopythium sp., and a wide range of plant species. The lower limit of linear detection using purified DNA was 200 to 300 fg of DNA in all pathogen RPA assays. Six different extraction buffers were tested for use during plant tissue maceration and the assays were validated in the field by collecting 222 symptomatic plant samples from over 50 different hosts. Only 56 samples were culture positive for Phytophthora spp. whereas 91 were positive using the Phytophthora genus-specific RPA test and a TaqMan real-time PCR assay. A technique for the generation of sequencing templates from positive RPA amplifications to confirm species identification was also developed. These RPA assays have added benefits over traditional technologies because they are rapid (results can be obtained in as little as 15 min), do not require DNA extraction or extensive training to complete, use less expensive portable equipment than PCR-based assays, and are significantly more specific than current immunologically based methods. This should provide a rapid, field-deployable capability for pathogen detection that will facilitate point-of-sample collection processing

  12. Application of the comet assay in studies of programmed cell death (PCD) in plants



    Programmed cell death (PCD) in plants is an intensively investigated process. One of the main characteristics of PCD in both animal and plant organisms is the non-random, internucleosomal fragmentation of nuclear DNA, usually analysed using total DNA gel electrophoresis or TUNEL method. In this paper we present application of the "comet assay" (Single Cell Gel Electrophoresis) for detection of nDNA degradation in studies of PCD during plant life cycle. We analyzed three types of tissue: anthe...

  13. Purification and characterization of oligonucleotide binding (OB)-fold protein from medicinal plant Tinospora cordifolia. (United States)

    Amir, Mohd; Haque, Md Anzarul; Wahiduzzaman; Dar, Mohammad Aasif; Islam, Asimul; Ahmad, Faizan; Hassan, Md Imtaiyaz


    The oligonucleotide binding fold (OB-fold) is a small structural motif present in many proteins. It is originally named for its oligonucleotide or oligosaccharide binding properties. These proteins have been identified as essential for replication, recombination and repair of DNA. We have successfully purified a protein contains OB-fold from the stem of Tinospora cordifolia, a medicinal plants of north India. Stems were crushed and centrifuged, and fraction obtained at 60% ammonium sulphate was extensively dialyzed and applied to the weak anion exchange chromatography on Hi-Trap DEAE-FF in 50mM Tris-HCl buffer at pH 8.0. Eluted fractions were concentrated and applied to gel filtration column to get pure protein. We observed a single band of 20-kDa on SDS-PAGE. Finally, the protein was identified as OB-fold by MALDI-TOF. The purified OB-fold protein was characterized for its secondary structural elements using circular dichroism (CD) in the far-UV region. Generally the OB-fold has a characteristic feature as five-stranded beta-sheet coiled to form a closed beta- barrel. To estimate its chemical stability, guanidinium chloride-induced denaturation curve was followed by observing changes in the far-UV CD as a function of the denaturant concentration. Analysis of this denaturation curve gave values of 8.90±0.25kcalmol(-1) and 3.78±0.18M for ΔGD° (Gibbs free energy change at 25°C) and Cm (midpoint of denaturation), respectively. To determine heat stability parameters of OB-fold protein, differential scanning calorimetry was performed. Calorimetric values of ΔGD°, Tm (midpoint of denaturation), ΔHm (enthalpy change at Tm), and ΔCp (constant-pressure heat capacity change) are 9.05±0.27kcalmol(-1), 85.2±0,3°C, 105±4kcalmol(-1) and 1.6±0.08kcalmol(-1)K(-1). This is the first report on the isolation, purification and characterization of OB-fold protein from a medicinal plant T. cordifolia.

  14. Optimization of Ammonium Sulfate Concentration for Purification of Colorectal Cancer Vaccine Candidate Recombinant Protein GA733-FcK Isolated from Plants. (United States)

    Park, Se-Ra; Lim, Chae-Yeon; Kim, Deuk-Su; Ko, Kisung


    A protein purification procedure is required to obtain high-value recombinant injectable vaccine proteins produced in plants as a bioreactor. However, existing purification procedures for plant-derived recombinant proteins are often not optimized and are inefficient, with low recovery rates. In our previous study, we used 25-30% ammonium sulfate to precipitate total soluble proteins (TSPs) in purification process for recombinant proteins from plant leaf biomass which has not been optimized. Thus, the objective in this study is to optimize the conditions for plant-derived protein purification procedures. Various ammonium sulfate concentrations (15-80%) were compared to determine their effects on TSPs yield. With 50% ammonium sulfate, the yield of precipitated TSP was the highest, and that of the plant-derived colorectal cancer-specific surface glycoprotein GA733 fused to the Fc fragment of human IgG tagged with endoplasmic reticulum retention signal KDEL (GA733(P)-FcK) protein significantly increased 1.8-fold. SDS-PAGE analysis showed that the purity of GA733(P)-FcK protein band appeared to be similar to that of an equal dose of mammalian-derived GA733-Fc (GA733(M)-Fc). The binding activity of purified GA733(P)-FcK to anti-GA733 mAb was as efficient as the native GA733(M)-Fc. Thus, the purification process was effectively optimized for obtaining a high yield of plant-derived antigenic protein with good quality. In conclusion, the purification recovery rate of large quantities of recombinant protein from plant expression systems can be enhanced via optimization of ammonium sulfate concentration during downstream processes, thereby offering a promising solution for production of recombinant GA733-Fc protein in plants.

  15. Investigations of plant-derived products with the in vitro comet assay

    Directory of Open Access Journals (Sweden)

    Luc Verschaeve


    It is impossible to perform a wide range of tests for screening purposes and often only the Ames assay is performed, which is insufficient. Furthermore, this test is most probably not the best choice when plant extracts need to be tested, because they often contain high amounts of histidine and have antibacterial properties. We have participated in many screening programs of medicinal plants and used different genotoxicity tests (mainly Ames assay, Vitotox test, micronucleus test and comet assay. Or results revealed that a combination of the Vitotox test and comet assay provides sufficiently reliable data with respect to genotoxicity as well as antigenotoxicity. This holds true for the testing of (medicinal plant extracts but also other plant derived products, for example those aimed at identifying novel TB chemotherapeutic drugs. The investigation of smoke and smoke compounds as enhancers of seed germination and smoke treated plants provides another example in which the comet assay proved to be valuable in the assessment of potential adverse health effects resulting from such treatment.

  16. Serine transhydroxymethylase: a simplified radioactive assay; purification and stabilization of enzyme activity employing Affi-Gel Blue. (United States)

    Braman, J C; Black, M J; Mangum, J H


    An improved radioactive assay has been developed for serine transhydroxymethylase. This assay involves the direct measurement of the [14C]HCHO which is generated when [3- 14C]-serine is employed as the substrate. The new assay eliminates the need for a solvent extraction of a [14C]HCHO-dimedon adduct which is the basis of the assay devised by Taylor and Weissbach. The enzyme has been purified employing Affi-Gel Blue. The purified enzyme retains full activity when bound to this affinity chromatography matrix and can be stored in this state at 4 degrees indefinitely.

  17. Purification and Structural Analysis of a Selective Toxin Fraction Produced by the Plant Pathogen Setosphaeria turcica

    Institute of Scientific and Technical Information of China (English)

    ZHANG Li-hui; DONG Jin-gao; WANG Chao-hua; LI Zheng-ping


    Thirteen fractions from the pathogenic plant fungus Setosphaeria turcica race 1 were separated and collected using high performance liquid chromatography (HPLC). Their toxic activities were assayed through leaf puncturing on corn differentials (OH43, OH43Htl, OH43Ht2, and OH43HtN), and the results revealed that eight fractions were toxic and fraction 6 was specifically toxic to OH43Htl, which could be taken as a gene-selective toxin fraction. Fraction 6 was finely purified via HPLC and condensed by freeze desiccation. Its chemical structure was analyzed with EI-MS, IR, HMBC, 1H-NMR, and two-dimensional NMR. The results suggested that fraction 6 contained an unsaturated double bond, carbonyl and methylene groups with molecular weight of 142.

  18. Use of a tandem affinity purification assay to detect interactions between West Nile and dengue viral proteins and proteins of the mosquito vector. (United States)

    Colpitts, Tonya M; Cox, Jonathan; Nguyen, Annie; Feitosa, Fabiana; Krishnan, Manoj N; Fikrig, Erol


    West Nile and dengue viruses are (re)emerging mosquito-borne flaviviruses that cause significant morbidity and mortality in man. The identification of mosquito proteins that associate with flaviviruses may provide novel targets to inhibit infection of the vector or block transmission to humans. Here, a tandem affinity purification (TAP) assay was used to identify 18 mosquito proteins that interact with dengue and West Nile capsid, envelope, NS2A or NS2B proteins. We further analyzed the interaction of mosquito cadherin with dengue and West Nile virus envelope protein using co-immunoprecipitation and immunofluorescence. Blocking the function of select mosquito factors, including actin, myosin, PI3-kinase and myosin light chain kinase, reduced both dengue and West Nile virus infection in mosquito cells. We show that the TAP method may be used in insect cells to accurately identify flaviviral-host protein interactions. Our data also provides several targets for interrupting flavivirus infection in mosquito vectors.

  19. Plant compounds enhance the assay sensitivity for detection of active Bacillus cereus toxin. (United States)

    Rasooly, Reuven; Hernlem, Bradley; He, Xiaohua; Friedman, Mendel


    Bacillus cereus is an important food pathogen, producing emetic and diarrheal syndromes, the latter mediated by enterotoxins. The ability to sensitively trace and identify this active toxin is important for food safety. This study evaluated a nonradioactive, sensitive, in vitro cell-based assay, based on B. cereus toxin inhibition of green fluorescent protein (GFP) synthesis in transduced monkey kidney Vero cells, combined with plant extracts or plant compounds that reduce viable count of B. cereus in food. The assay exhibited a dose dependent GFP inhibition response with ~25% inhibition at 50 ng/mL toxin evaluated in culture media or soy milk, rice milk or infant formula, products associated with food poisonings outbreak. The plant extracts of green tea or bitter almond and the plant compounds epicatechin or carvacrol were found to amplify the assay response to ~90% inhibition at the 50 ng/mL toxin concentration greatly increasing the sensitivity of this assay. Additional studies showed that the test formulations also inhibited the growth of the B. cereus bacteria, likely through cell membrane disruption. The results suggest that the improved highly sensitive assay for the toxin and the rapid inactivation of the pathogen producing the toxin have the potential to enhance food safety.

  20. Plant Compounds Enhance the Assay Sensitivity for Detection of Active Bacillus cereus Toxin

    Directory of Open Access Journals (Sweden)

    Reuven Rasooly


    Full Text Available Bacillus cereus is an important food pathogen, producing emetic and diarrheal syndromes, the latter mediated by enterotoxins. The ability to sensitively trace and identify this active toxin is important for food safety. This study evaluated a nonradioactive, sensitive, in vitro cell-based assay, based on B. cereus toxin inhibition of green fluorescent protein (GFP synthesis in transduced monkey kidney Vero cells, combined with plant extracts or plant compounds that reduce viable count of B. cereus in food. The assay exhibited a dose dependent GFP inhibition response with ~25% inhibition at 50 ng/mL toxin evaluated in culture media or soy milk, rice milk or infant formula, products associated with food poisonings outbreak. The plant extracts of green tea or bitter almond and the plant compounds epicatechin or carvacrol were found to amplify the assay response to ~90% inhibition at the 50 ng/mL toxin concentration greatly increasing the sensitivity of this assay. Additional studies showed that the test formulations also inhibited the growth of the B. cereus bacteria, likely through cell membrane disruption. The results suggest that the improved highly sensitive assay for the toxin and the rapid inactivation of the pathogen producing the toxin have the potential to enhance food safety.

  1. Application of the comet assay in studies of programmed cell death (PCD in plants

    Directory of Open Access Journals (Sweden)

    Maria Charzyńska


    Full Text Available Programmed cell death (PCD in plants is an intensively investigated process. One of the main characteristics of PCD in both animal and plant organisms is the non-random, internucleosomal fragmentation of nuclear DNA, usually analysed using total DNA gel electrophoresis or TUNEL method. In this paper we present application of the "comet assay" (Single Cell Gel Electrophoresis for detection of nDNA degradation in studies of PCD during plant life cycle. We analyzed three types of tissue: anther tapetum, endosperm and mesophyll which were prepared in different ways to obtain a suspension of viable cells (without cell walls. The comet assay gives a possibility of examination of the nDNA degradation in individual cell. This method is significant for studies of the plant tissue differentiation and senescence especially in the cases when it is not possible to isolate large number of cells at the same developmental stage.

  2. Purification of recombinant human and Drosophila septin hexamers for TIRF assays of actin-septin filament assembly. (United States)

    Mavrakis, M; Tsai, F-C; Koenderink, G H


    Septins are guanine nucleotide-binding proteins that are conserved from fungi to humans. Septins assemble into heterooligomeric complexes and higher-order structures with key roles in various cellular functions including cell migration and division. The mechanisms by which septins assemble and interact with other cytoskeletal elements like actin remain elusive. A powerful approach to address this question is by cell-free reconstitution of purified cytoskeletal proteins combined with fluorescence microscopy. Here, we describe procedures for the purification of recombinant Drosophila and human septin hexamers from Escherichia coli and reconstitution of actin-septin coassembly. These procedures can be used to compare assembly of Drosophila and human septins and their coassembly with the actin cytoskeleton by total internal reflection fluorescence microscopy.

  3. Plant uptake-assisted round-the-clock photocatalysis for complete purification of aquaculture wastewater using sunlight. (United States)

    Bian, Zhenfeng; Cao, Fenglei; Zhu, Jian; Li, Hexing


    A novel reactor equipped with solar batteries, Bi2O3/TiO2 film photocatalyst, and celery plant was designed and used for purification of aquaculture wastewater. The Bi2O3/TiO2 film photocatalyst started photocatalytic degradation of organonitrogen compounds under irradiation of sunlight. Meanwhile, the solar batteries absorbed and converted excess sunlight into electric energy and then started UV lamps at night, leading to round-the-clock photocatalysis. Subsequently, the inorganic nitrogen species including NH4(+), NO2(-), and NO3(-) resulting from photocatalytic degradation of the organonitrogen compounds could subsequently be uptaken by the celery plant as the fertilizer to reduce the secondary pollution. It was found that, after 24 h circulation, both organonitrogen compounds and NO2(-) species were completely removed, while NH4(+) and NO3(-) contents also decreased by 30% and 50%, respectively. The reactor could be used repetitively, showing a good potential in practical application.

  4. Application of the chloramphenicol acetyltransferase (CAT) diffusion assay to transgenic plant tissues. (United States)

    Peach, C; Velten, J


    Chloramphenicol acetyltransferase (CAT) activity was quantified in crude extracts from tobacco callus tissues using a modification of a previously reported diffusion assay. We describe here the alterations necessary in applying this rapid and simple assay procedure to plant materials. Due to the high concentration of nonspecific oxidases present in most plant tissues, some type of protective agent is required to maintain enzyme activity. We have tested beta-mercaptoethanol, cysteine, dithiothreitol, ascorbic acid and polyvinyl pyrrolidone as protective agents within the initial extraction buffer. We also investigated the effect of heat (60 degrees C, 10 min) and 5 mM EDTA on CAT activity. The highest CAT activity was obtained using 5 mM cysteine plus 5 mM EDTA in 40 mM Tris-HCl (pH 7.8) as the initial extraction buffer followed by a heat treatment. Using this buffer, CAT activity was stable on ice for more than two hours. In our hands, total acetyl-coenzyme A concentration within the assay mixture was found to be saturating at 250 microM and the Km determined to be 100 microM. Assays performed using the same crude plant extract indicate that 1) duplicate assays show less than 1.5% variation in activities and 2) CAT activity increases linearly with respect to volume of extract used.

  5. Development of rapid isothermal amplification assays for Phytophthora species from plant tissue (United States)

    Several isothermal amplification techniques recently have been developed that are tolerant of inhibitors present in many plant extracts, which can reduce the need for obtaining purified DNA for running diagnostic assays. One such commercially available technique that has similarities with real time ...

  6. Toxicity of selected plant volatiles in microbial and mammalian short-term assays

    NARCIS (Netherlands)

    Stammati, A.; Bonsi, P.; Zucco, F.; Moezelaar, R.; Alakomi, H.L.; von Wright, A.


    In this study, several short-term microbial and mammalian in vitro assays were used to evaluate cytotoxicity and genotoxicity of four plant volatiles showing antifungal activity: cinnamaldehyde, carvacrol, thymol and S(+)-carvone. All inhibited viability and proliferation of Hep-2 cells in a dose-de

  7. Toxicity of selected plant volatiles in microbial and mammalian short-term assays

    NARCIS (Netherlands)

    Stammati, A.; Bonsi, P.; Zucco, F.; Moezelaar, R.; Alakomi, H.L.; Wright, von A.


    In this study, several short-term microbial and mammalian in vitro assays were used to evaluate cytotoxicity and genotoxicity of four plant volatiles showing antifungal activity: cinnamaldehyde, carvacrol, thymol and S(+)-carvone. All inhibited viability and proliferation of Hep-2 cells in a dose-de

  8. The comet assay in higher terrestrial plant model: Review and evolutionary trends. (United States)

    Lanier, Caroline; Manier, Nicolas; Cuny, Damien; Deram, Annabelle


    The comet assay is a sensitive technique for the measurement of DNA damage in individual cells. Although it has been primarily applied to animal cells, its adaptation to higher plant tissues significantly extends the utility of plants for environmental genotoxicity research. The present review focuses on 101 key publications and discusses protocols and evolutionary trends specific to higher plants. General consensus validates the use of the percentage of DNA found in the tail, the alkaline version of the test and root study. The comet protocol has proved its effectiveness and its adaptability for cultivated plant models. Its transposition in wild plants thus appears as a logical evolution. However, certain aspects of the protocol can be improved, namely through the systematic use of positive controls and increasing the number of nuclei read. These optimizations will permit the increase in the performance of this test, namely when interpreting mechanistic and physiological phenomena.

  9. Membrane Distillation and Applications for Water Purification in Thermal Cogeneration. Pilot Plant Trials

    Energy Technology Data Exchange (ETDEWEB)

    Kullab, Alaa; Martin, Andrew


    Water treatment is an important auxiliary process in all thermal cogeneration plants. In this context membrane distillation (MD) is a novel technology that is potentially advantageous to technologies like reverse osmosis in the following ways: ability to utilize low-grade heat; reduced sensitivity to fluctuations in pH or salt concentrations; and lower capital and operation and maintenance costs (assumed in the case of fully-developed technology only). This research is a continuation of a Varmeforsk prestudy (report no. 909) and encompasses field trials at Idbaecken Combined Heat and Power (CHP) Facility (Nykoeping). Target groups for this study include environmental engineers with particular interest in emerging water purification technologies. The test rig consisted of a five-module MD unit capable of producing 1-2 m3/day purified water. District heating supply was employed for heating; feed stocks include municipal water and flue gas condensate. Field trials can be divided into three phases: (1) parametric study of yield; (2) long term operation with municipal water as feed stock; and (3) evaluation of flue gas condensate as a feed stock. Testing commenced in the beginning of April 2006. The performance of MD concerning production rate is highly dependent on the feed stock temperature, flow rate and temperature difference across the membrane. Initial results for municipal water feed stocks showed that product water fluxes were in line with previous experiments, thus confirming the findings made in the prestudy. Connecting several MD modules in series has the advantage of reducing the electrical energy consumption needed for recirculation; the penalty comes in less efficient operation from flux point of view. This is more critical in the case of low flow rates, and hence much careful design studies are needed to optimize the system. Regarding the long term performance, the test period lasted for 13 days on a continuous operation basis before the first flux

  10. Membrane Distillation and Applications for Water Purification in Thermal Cogeneration. Pilot Plant Trials

    Energy Technology Data Exchange (ETDEWEB)

    Kullab, Alaa; Martin, Andrew


    Water treatment is an important auxiliary process in all thermal cogeneration plants. In this context membrane distillation (MD) is a novel technology that is potentially advantageous to technologies like reverse osmosis in the following ways: ability to utilize low-grade heat; reduced sensitivity to fluctuations in pH or salt concentrations; and lower capital and operation and maintenance costs (assumed in the case of fully-developed technology only). This research is a continuation of a Varmeforsk prestudy (report no. 909) and encompasses field trials at Idbaecken Combined Heat and Power (CHP) Facility (Nykoeping). Target groups for this study include environmental engineers with particular interest in emerging water purification technologies. The test rig consisted of a five-module MD unit capable of producing 1-2 m3/day purified water. District heating supply was employed for heating; feed stocks include municipal water and flue gas condensate. Field trials can be divided into three phases: (1) parametric study of yield; (2) long term operation with municipal water as feed stock; and (3) evaluation of flue gas condensate as a feed stock. Testing commenced in the beginning of April 2006. The performance of MD concerning production rate is highly dependent on the feed stock temperature, flow rate and temperature difference across the membrane. Initial results for municipal water feed stocks showed that product water fluxes were in line with previous experiments, thus confirming the findings made in the prestudy. Connecting several MD modules in series has the advantage of reducing the electrical energy consumption needed for recirculation; the penalty comes in less efficient operation from flux point of view. This is more critical in the case of low flow rates, and hence much careful design studies are needed to optimize the system. Regarding the long term performance, the test period lasted for 13 days on a continuous operation basis before the first flux

  11. Detection of anti-tuberculosis activity in some folklore plants by radiometric BACTEC assay. (United States)

    Gupta, V K; Shukla, C; Bisht, G R S; Saikia, D; Kumar, S; Thakur, R L


    The anti-tubercular drugs are less effective because of the emergence of multi-drug resistant (MDR) and extensively drug resistant (XDR) strains of M. tuberculosis, so plants being an alternative source of anti-microbial compounds. The aim of this study was to investigate anti-tuberculosis potential of the plants using Mycobacterium smegmatis as a rapid screening model for detection of anti-mycobacterial activity and further to evaluate the active plants for anti-tuberculosis activity against M. tuberculosis using radiometric BACTEC assay. The 15 plants were screened for anti-mycobacterial activity against M. smegmatis by the disk diffusion assay. The ethanolic extracts of Mallotus philippensis, Vitex negundo, Colebrookea oppositifolia, Rumex hastatus, Mimosa pudica, Kalanchoe integra and Flacourtia ramontchii were active against M. smegmatis in primary screening. The anti-tuberculosis potential was identified in the leaves extracts of Mallotus philippensis by radiometric BACTEC assay. The ethanolic extract of M. philippensis showed anti-tuberculosis activity against virulent and avirulent strains of M. tuberculosis H(37) Rv and M. tuberculosis H(37) Ra with minimum inhibitory concentration 0·25 and 0·125 mg ml(-1), respectively. The inhibition in growth index values of M. tuberculosis was observed in the presence of ethyl acetate fraction at a minimum concentration of 0·05 mg ml(-1). We found that BACTEC radiometric assay is a valuable method for detection of anti-tuberculosis activity of the plant extracts. The results indicate that ethanolic extract and ethyl acetate fraction of M. philippensis exhibited significant anti-mycobacterial activity against M. tuberculosis. These findings provide scientific evidence to support the traditional medicinal uses of M. philippensis and indicate a promising potential of this plant for the development of anti-tuberculosis agent. © 2010 The Authors. Letters in Applied Microbiology © 2010 The Society for Applied

  12. Molecular characterization of the bacterial communities in the different compartments of a full-scale reverse-osmosis water purification plant

    NARCIS (Netherlands)

    Bereschenko, L.A.; Heilig, G.H.J.; Nederlof, M.M.; Loosdracht, van M.C.M.; Stams, A.J.M.; Euverink, G.J.W.


    The origin, structure, and composition of biofilms in various compartments of an industrial full-scale reverse-osmosis (RO) membrane water purification plant were analyzed by molecular biological methods. Samples were taken when the RO installation suffered from a substantial pressure drop and decre

  13. Molecular Characterization of the Bacterial Communities in the Different Compartments of a Full-Scale Reverse-Osmosis Water Purification Plant

    NARCIS (Netherlands)

    Bereschenko, L.A.; Heilig, G.H.J.; Nederlof, M.M.; Loosdrecht, M.C.M. van; Stams, A.J.M.; Euverink, G.J.W.


    The origin, structure, and composition of biofilms in various compartments of an industrial full-scale reverse-osmosis (RO) membrane water purification plant were analyzed by molecular biological methods. Samples were taken when the RO installation suffered from a substantial pressure drop and decre

  14. One-step non-chromatography purification of a low abundant fucosylated protein from complex plant crude extract

    Directory of Open Access Journals (Sweden)

    Lindsay Arnold


    Full Text Available Effective methods for isolation and purification of glycoproteins and other glycoconjugates are important to biopharmaceutical industry and diagnostic industry. They are also critical to an emerging field of glycoproteomics. In this work, we applied the newly-developed affinity ligand, a fusion protein of elastic like polymer (ELP and a bacterial lectin, in an affinity precipitation process to purify soybean peroxidase (SBP based on the presence of fucoseon the protein surface. We addressed, in particular, the challenge of purifying a low abundant protein from a complex dilute crude plant extract. The novel affinity precipitation developed in this work was very promising. One step binding and precipitation resulted in >95% recovery yield directly from crude extract and a 22.7 fold purification, giving a specific activity of 420 U/mg. The SBP isolated using this affinity precipitation meets or exceeds the quality specifications of reagent grade products by Sigma. We showed that the recovery yield had a strong dependence on the molar ratio of ligand to target fucosylated protein, with a ratio of three giving nearly full recovery, which could be predicted based on the total fucose content per protein molecule and the number of binding site per ligand molecule. We additionally developed a method of ligand regeneration and investigated its reuse. A simple wash with pH buffer was shown to be effective to regenerate the binding capacity for the ligand, and the ligand could be used for 10 times, giving an averaged 80% isolation yield based on initial input of soybean peroxidase. Taken together, an effective method of affinity precipitation was developed, which could be used to enrich a low abundant target glycoprotein from a complex mixture with a high recovery yield. The high selectivity for fucosylated protein and its ease of operation make this method particularly useful for purification of low abundant glycoprotein from natural sources. This work

  15. A modified MS2 bacteriophage plaque reduction assay for the rapid screening of antiviral plant extracts

    Directory of Open Access Journals (Sweden)

    Ian Cock


    Full Text Available Introduction: Traditional methods of screening plant extracts and purified components for antiviral activity require up to a week to perform, prompting the need to develop more rapid quantitative methods to measure the ability of plant based preparations to block viral replication. We describe an adaption of an MS2 plaque reduction assay for use in S. aureus. Results: MS2 bacteriophage was capable of infecting and replicating in B. cereus, S. aureus and F+ E. coli but not F- E. coli. Indeed, both B. cereus and S. aureus were more sensitive to MS2 induced lysis than F+ E. coli. When MS2 bacteriophage was mixed with Camellia sinensis extract (1 mg/ml, Scaevola spinescens extract (1 mg/ml or Aloe barbadensis juice and the mixtures inoculated into S. aureus, the formation of plaques was reduced to 8.9 ± 3.8%, 5.4 ± 2.4% and 72.7 ± 20.9% of the untreated MS2 control values respectively. Conclusions: The ability of the MS2 plaque reduction assay to detect antiviral activity in these known antiviral plant preparations indicates its suitability as an antiviral screening tool. An advantage of this assay compared with traditionally used cytopathic effect reduction assays and replicon based assays is the more rapid acquisition of results. Antiviral activity was detected within 24 h of the start of testing. The MS2 assay is also inexpensive and non-pathogenic to humans making it ideal for initial screening studies or as a simulant for pathogenic viruses.

  16. Purification and assay of cell-invasive form of calmodulin-sensitive adenylyl cyclase from Bordetella pertussis

    Energy Technology Data Exchange (ETDEWEB)

    Masure, H.R.; Donovan, M.G.; Storm, D.R.


    An invasive form of the CaM-sensitive adenylyl cyclase from Bordetella pertussis can be isolated from bacterial culture supernatants. This isolation is achieved through the use of QAE-Sephadex anion-exchange chromatography. It has been demonstrated that the addition of exogenous Ca{sup 2}{sup +} to the anion-exchange gradient buffers will affect elution from the column and will thereby affect the isolation of invasive adenylyl cyclase. This is probably due to a Ca2(+)-dependent interaction of the catalytic subunit with another component in the culture supernatant. Two peaks of adenylyl cyclase activity are obtained. The Pk1 adenylyl cyclase preparation is able to cause significant increases in intracellular cAMP levels in animal cells. This increase occurs rapidly and in a dose-dependent manner in both N1E-115 mouse neuroblastoma cells and human erythrocytes. The Pk2 adenylyl cyclase has catalytic activity but is not cell invasive. This material can serve, therefore, as a control to ensure that the cAMP which is measured is, indeed, intracellular. A second control is to add exogenous CaM to the Pk1 adenylyl cyclase preparation. The 45-kDa catalytic subunit-CaM complex is not cell invasive. Although the mechanism for membrane translocation of the adenylyl cyclase is unknown, there is evidence that the adenylyl cyclase enters animal cells by a mechanism distinct from receptor-mediated endocytosis. Calmodulin-sensitive adenylyl cyclase activity can be removed from preparations of the adenylyl cyclase that have been subjected to SDS-polyacrylamide gel electrophoresis. This property of the enzyme has enabled purification of the catalytic subunit to apparent homogeneity. The purified catalytic subunit from culture supernatants has a predicted molecular weight of 45,000. This polypeptide interacts directly with Ca{sup 2}{sup +} and this interaction may be important for its invasion into animal cells.

  17. Production, purification and assay of thrombopoietin. Progress report, August 1, 1975--July 31, 1976. [X radiation, /sup 35/S

    Energy Technology Data Exchange (ETDEWEB)

    McDonald, T.P.


    Experiments were conducted which indicate that thrombopoietin can be detected by both a bioassay and an immunoassay in sera of patients with various platelet production disorders. Other studies have shown that the kidney appears to have a vital role in thrombopoietin production; the mechanism of how platelet-specific antisera causes thrombocytopenia has been investigated. Also, an investigation has been made into the preparation of assay mice and the different methods for the measurement of thrombopoietin. These studies indicate that mice in rebound-thrombocytosis are more sensitive to exogenous TSF than normal or platelet transfused mice. Also, % /sup 35/S incorporation into platelets of assay mice is the most sensitive measurement of thrombopoiesis. The effects of hypoxia on platelet production was also investigated along with the release of thrombopoietin in animals exposed to RAMPS and whole-body x-irradiation.

  18. Purification and Characterization of Two Voltage-Dependent Anion Channel Isoforms from Plant Seeds1 (United States)

    Abrecht, Helge; Wattiez, Ruddy; Ruysschaert, Jean-Marie; Homblé, Fabrice


    Mitochondria were isolated from imbibed seeds of lentil (Lens culinaris) and Phaseolus vulgaris. We copurified two voltage-dependent anion channel from detergent solubilized mitochondria in a single purification step using hydroxyapatite. The two isoforms from P. vulgaris were separated by chromatofocusing chromatography in 4 m urea without any loss of channel activity. Channel activity of each isoform was characterized upon reconstitution into diphytanoyl phosphatidylcholine planar lipid bilayers. Both isoforms form large conductance channels that are slightly anion selective and display cation selective substates. PMID:11080295

  19. Heat-precipitation allows the efficient purification of a functional plant-derived malaria transmission-blocking vaccine candidate fusion protein. (United States)

    Beiss, Veronique; Spiegel, Holger; Boes, Alexander; Kapelski, Stephanie; Scheuermayer, Matthias; Edgue, Gueven; Sack, Markus; Fendel, Rolf; Reimann, Andreas; Schillberg, Stefan; Pradel, Gabriele; Fischer, Rainer


    Malaria is a vector-borne disease affecting more than two million people and accounting for more than 600,000 deaths each year, especially in developing countries. The most serious form of malaria is caused by Plasmodium falciparum. The complex life cycle of this parasite, involving pre-erythrocytic, asexual and sexual stages, makes vaccine development cumbersome but also offers a broad spectrum of vaccine candidates targeting exactly those stages. Vaccines targeting the sexual stage of P. falciparum are called transmission-blocking vaccines (TBVs). They do not confer protection for the vaccinated individual but aim to reduce or prevent the transmission of the parasite within a population and are therefore regarded as an essential tool in the fight against the disease. Malaria predominantly affects large populations in developing countries, so TBVs need to be produced in large quantities at low cost. Combining the advantages of eukaryotic expression with a virtually unlimited upscaling potential and a good product safety profile, plant-based expression systems represent a suitable alternative for the production of TBVs. We report here the high level (300 μg/g fresh leaf weight (FLW)) transient expression in Nicotiana benthamiana leaves of an effective TBV candidate based on a fusion protein F0 comprising Pfs25 and the C0-domain of Pfs230, and the implementation of a simple and cost-effective heat treatment step for purification that yields intact recombinant protein at >90% purity with a recovery rate of >70%. The immunization of mice clearly showed that antibodies raised against plant-derived F0 completely blocked the formation of oocysts in a malaria transmission-blocking assay (TBA) making F0 an interesting TBV candidate or a component of a multi-stage malaria vaccine cocktail.

  20. Sensitivity of two in vitro assays for evaluating plant activity against the infective stage of Haemonchus contortus strains. (United States)

    Al-Rofaai, A; Rahman, W A; Abdulghani, Mahfoudh


    The sensitivity of larval paralysis assay (LPA) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide-formazan (MTT-formazan) assay was compared to evaluate the anthelmintic activity of plant extracts. In this study, the methanolic extract of Azadirachta indica (neem) was evaluated for its activity against the infective-stage larvae (L(3)) of susceptible and resistant Haemonchus contortus strains using the two aforementioned assays. In both in vitro assays, the same serial concentrations of the extract were used, and the median lethal concentrations were determined to compare the sensitivity of both assays. The results revealed a significant difference (P formazan assay. The MTT-formazan assay is more feasible for practical applications because it measured the L(3) mortality more accurately than LPA. This study may help find a suitable assay for investigating the anthelmintic activity of plant extracts against trichostrongylid nematodes.

  1. Process for purification of monoclonal antibody expressed in transgenic Lemna plant extract using dextran-coated charcoal and hexamer peptide affinity resin. (United States)

    Naik, Amith D; Menegatti, Stefano; Reese, Hannah R; Gurgel, Patrick V; Carbonell, Ruben G


    The production of therapeutic proteins using transgenic plants offers several advantages, including low production cost, absence of human pathogens, presence of glycosylation mechanisms, and the ability to fold complex therapeutic proteins into their proper conformation. However, impurities such as phenolic compounds and pigments encountered during purification are quite different from those faced during purification from mammalian cell culture supernatants. This paper deals with the development of a pretreatment and affinity separation process for the purification of a monoclonal antibody from transgenic Lemna plant extract. A pretreatment step is described using dextran-coated charcoal for the removal of pigments and phenolic compounds without reducing the antibody concentration. Then, the peptide affinity ligand HWRGWV coupled to a commercial polymethacrylate resin is used for the capture and purification of MAb from the pretreated plant extract. The final yield and purity of the MAb obtained were 90% and 96% respectively. The performance of the hexamer peptide resin after the pretreatment step was found to be similar to that obtained with a commercial Protein A resin.

  2. Plant Growth and Water Purification of Porous Vegetation Concrete Formed of Blast Furnace Slag, Natural Jute Fiber and Styrene Butadiene Latex

    Directory of Open Access Journals (Sweden)

    Hwang-Hee Kim


    Full Text Available The purpose of this study is to investigate porous vegetation concrete formed using the industrial by-products blast furnace slag powder and blast furnace slag aggregates. We investigated the void ratio, compressive strength, freeze–thaw resistance, plant growth and water purification properties using concretes containing these by-products, natural jute fiber and latex. The target performance was a compressive strength of ≥12 MPa, a void ratio of ≥25% and a residual compressive strength of ≥80% following 100 freeze–thaw cycles. Using these target performance metrics and test results for plant growth and water purification, an optimal mixing ratio was identified. The study characterized the physical and mechanical properties of the optimal mix, and found that the compressive strength decreased compared with the default mix, but that the void ratio and the freeze–thaw resistance increased. When latex was used, the compressive strength, void ratio and freeze–thaw resistance all improved, satisfying the target performance metrics. Vegetation growth tests showed that plant growth was more active when the blast furnace slag aggregate was used. Furthermore, the use of latex was also found to promote vegetation growth, which is attributed to the latex forming a film coating that suppresses leaching of toxic components from the cement. Water purification tests showed no so significant differences between different mixing ratios; however, a comparison of mixes with and without vegetation indicated improved water purification in terms of the total phosphorus content when vegetation had been allowed to grow.

  3. Total Measurement Uncertainty for the Plutonium Finishing Plant (PFP) Segmented Gamma Scan Assay System

    CERN Document Server

    Fazzari, D M


    This report presents the results of an evaluation of the Total Measurement Uncertainty (TMU) for the Canberra manufactured Segmented Gamma Scanner Assay System (SGSAS) as employed at the Hanford Plutonium Finishing Plant (PFP). In this document, TMU embodies the combined uncertainties due to all of the individual random and systematic sources of measurement uncertainty. It includes uncertainties arising from corrections and factors applied to the analysis of transuranic waste to compensate for inhomogeneities and interferences from the waste matrix and radioactive components. These include uncertainty components for any assumptions contained in the calibration of the system or computation of the data. Uncertainties are propagated at 1 sigma. The final total measurement uncertainty value is reported at the 95% confidence level. The SGSAS is a gamma assay system that is used to assay plutonium and uranium waste. The SGSAS system can be used in a stand-alone mode to perform the NDA characterization of a containe...

  4. Biological conversion assay using Clostridium phytofermentans to estimate plant feedstock quality

    Directory of Open Access Journals (Sweden)

    Lee Scott J


    Full Text Available Abstract Background There is currently considerable interest in developing renewable sources of energy. One strategy is the biological conversion of plant biomass to liquid transportation fuel. Several technical hurdles impinge upon the economic feasibility of this strategy, including the development of energy crops amenable to facile deconstruction. Reliable assays to characterize feedstock quality are needed to measure the effects of pre-treatment and processing and of the plant and microbial genetic diversity that influence bioconversion efficiency. Results We used the anaerobic bacterium Clostridium phytofermentans to develop a robust assay for biomass digestibility and conversion to biofuels. The assay utilizes the ability of the microbe to convert biomass directly into ethanol with little or no pre-treatment. Plant samples were added to an anaerobic minimal medium and inoculated with C. phytofermentans, incubated for 3 days, after which the culture supernatant was analyzed for ethanol concentration. The assay detected significant differences in the supernatant ethanol from wild-type sorghum compared with brown midrib sorghum mutants previously shown to be highly digestible. Compositional analysis of the biomass before and after inoculation suggested that differences in xylan metabolism were partly responsible for the differences in ethanol yields. Additionally, we characterized the natural genetic variation for conversion efficiency in Brachypodium distachyon and shrub willow (Salix spp.. Conclusion Our results agree with those from previous studies of lignin mutants using enzymatic saccharification-based approaches. However, the use of C. phytofermentans takes into consideration specific organismal interactions, which will be crucial for simultaneous saccharification fermentation or consolidated bioprocessing. The ability to detect such phenotypic variation facilitates the genetic analysis of mechanisms underlying plant feedstock quality.

  5. Purification Simulation With Vapor Permeation and Distillation-Adsorption In Bioethanol Plant

    Directory of Open Access Journals (Sweden)

    Misri Gozan


    Full Text Available High purity of Bioethanol is required in biofuel mixing with gasoline (EXX. In bioethanol production line, the azeotropic property of ethanol-water becomes the barrier for purification process. This study examined two bioethanol separation processes by support of simulation tools, Superpro Designer 9.0 software. Ethanol purity and a low costeconomical process were the major considerations. Purification method of vapor permeation membrane technology was compared with distillation-adsorption method. Data from previous lab experiments and some literatures were used. The results showed that distillation-adsorption method is more economical compared to vapor permeation technology. Payback period of the simulation is 3.9 years and 4.3 years to distillation adsorption and vapor permeation respectively with each IRR value is 20.23% and 17.89%. Initial investment value of vapor permeation is 9.6% higher than distillation method. Significant difference observed in operating costs, since more units involved in vapor permeation require more labors to operate.

  6. Rapid Point-of-Care Isothermal Amplification Assay for the Detection of Malaria without Nucleic Acid Purification. (United States)

    Modak, Sayli S; Barber, Cheryl A; Geva, Eran; Abrams, William R; Malamud, Daniel; Ongagna, Yhombi Serge Yvon


    Malaria remains one of the most prevalent infectious diseases and results in significant mortality. Isothermal amplification (loop-mediated isothermal amplification) is used to detect malarial DNA at levels of ~1 parasite/µL blood in ≤30 minutes without the isolation of parasite nucleic acid from subject's blood or saliva. The technique targets the mitochondrial cytochrome oxidase subunit 1 gene and is capable of distinguishing Plasmodium falciparum from Plasmodium vivax. Malarial diagnosis by the gold standard microscopic examination of blood smears is generally carried out only after moderate-to-severe symptoms appear. Rapid diagnostic antigen tests are available but generally require infection levels in the range of 200-2,000 parasites/µL for a positive diagnosis and cannot distinguish if the disease has been cleared due to the persistence of circulating antigen. This study describes a rapid and simple molecular assay to detect malarial genes directly from whole blood or saliva without DNA isolation.

  7. Effect of water purification process in radioactive content: analysis on small scale purification plants; Efecto del proceso de purificacion de agua en el contenido radiactivo: analisis en plantas purificadoras a pequena escala

    Energy Technology Data Exchange (ETDEWEB)

    Lopez del Rio, H.; Quiroga S, J. C.; Davila R, J. I.; Mireles G, F. [Universidad Autonoma de Zacatecas, Unidad Academica de Estudios Nucleares, Calle Cipres No. 10, Fracc. La Penuela, 98000, Zacatecas (Mexico)], e-mail:


    Water from small scale purification plants is a low cost alternative for consumers in comparison to the bottled commercial presentations. Because of its low cost per liter, the consumption of this product has increased in recent years, stimulating in turn the installation of purification systems for these small businesses. The purpose of this study was to estimate the efficiency of small scale purification systems located in the cities of Zacatecas and Guadalupe, Zacatecas, to reduce the radioactive content of water. It was measured the total alpha and beta activity in water samples of entry and exit to process, through the liquid scintillation technique. In general it was observed that the process is more efficient in removing alpha that beta activity. The fraction of total alpha activity removed varied between 27 and 100%, while between 0 and 77% of the total beta activity was removed by the analyzed plants. In all cases, the total radioactivity level was lower than the maximum permissible value settled by the official mexican standard for drinking water. (Author)

  8. Application of the Single Cell Gel Electrophoresis (SCGE) Assay to Genotoxicity Evaluation in Plants and Animals

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jin Kyu


    Application of the Single Cell Gel Electrophoresis (SCGE) Assay to Genotoxicity Evaluation in Plants and Animals. Recently, the importance of ionizing radiation and chemicals has been recognized since radio- and chemical therapy is directly related to the control of various diseases such as cancer. Radiation and the chemicals can cause biological damages while they have great applicability. It is of necessity to analyze rapidly, easily and accurately the biological effects, especially DNA damage due to those factors. Recently SCGE (single cell gel electrophoresis assay, alias comet assay) has been developed for the efficient evaluation of DNA damage. In this report, the comprehensive review will be given on the rationale, the technical applications and the advantages and shortcomings of SCGE assay. This method can be directly applied to study on toxicity, cancer, and aging in terms of the evaluation of DNA damages due to radiation and chemicals on human cellular level. It is also suggested that comet assay be used for testing genotoxicity of suspected substances, detecting irradiated foods, screening radioprotective candidates, and studying DNA repair process in various biological systems.

  9. Assays of dioxins and dioxin-like compounds in actually contaminated soils using transgenic tobacco plants carrying a recombinant mouse aryl hydrocarbon receptor-mediated β-glucuronidase reporter gene expression system. (United States)

    Inui, Hideyuki; Gion, Keiko; Utani, Yasushi; Wakai, Taketo; Kodama, Susumu; Eun, Heesoo; Kim, Yun-Seok; Ohkawa, Hideo


    The transgenic tobacco plant XD4V-26 carrying the recombinant mouse aryl hydrocarbon receptor XD4V-mediated β-glucuronidase (GUS) reporter gene expression system was used for assay of dioxins and dioxin-like compounds consisting of polychlorinated dibenzeno-p-dioxins, polychlorinated dibenzofurans, and coplanar polychlorinated biphenyls (Co-PCBs) in actually contaminated soils. The transgenic tobacco plant XD4V-26 showed a significant dose-dependent induced GUS activity when cultured on MS medium containing PCB126 [toxic equivalency factor (TEF) = 0.1]. In contrast, PCB169 and PCB180, which have 0.03 of TEF and unassigned TEF values, respectively, did not significantly induce GUS activity under the same conditions as with PCB126. When the tobacco plants were cultivated for up to 5 weeks on actually contaminated soils with dioxins and dioxin-like compounds collected from the periphery of an incinerator used for disposal of residential and industrial wastes, GUS activity in the leaves was dose-dependently increased. The plants clearly detected 360 pg-TEQ g(-1) of dioxins and dioxin-like compounds in this assay. There was a positive correlation between GUS activity and TEQ value of dioxins and dioxin-like compounds in the plants. This assay does not require any extraction and purification processes for the actually contaminated soil samples.

  10. Microbiological corrosion in aerobic and anaerobic waste purification plants: safety and efficiency problems

    Energy Technology Data Exchange (ETDEWEB)

    Perego, P. [ISTIC - Institute of Chemical and Process Engineering ``G.B. Bonino``, University of Genoa, Via Opera Pia 15-16145 Genoa (Italy); Fabiano, B. [ISTIC - Institute of Chemical and Process Engineering ``G.B. Bonino``, University of Genoa, Via Opera Pia 15-16145 Genoa (Italy); Pastorino, R. [ISTIC - Institute of Chemical and Process Engineering ``G.B. Bonino``, University of Genoa, Via Opera Pia 15-16145 Genoa (Italy); Randi, G. [ISTIC - Institute of Chemical and Process Engineering ``G.B. Bonino``, University of Genoa, Via Opera Pia 15-16145 Genoa (Italy)


    Carbon steels and stainless steels are largely employed in order to build up biological purification systems of urban and industrial waste. The former are applied in building up tanks, vessels and pipes; the latter are employed in building up operator machines and appliances. This paper presents a study of the corrosion behaviour of these materials in anaerobic and aerobic environments. Tests have been carried out by continuous immersion in a biologic liquid with mixed flora. The observation of samples taken at different times of exposure has given information of great interest of application, especially about the mechanisms of localized corrosion phenomena and about the behaviour of these materials in function. Besides the presented study shows original possibilities for the assessment of the corrosion phenomenon that can be adopted and developed further still. (orig.). With 7 figs., 4 tabs.

  11. In vitro antioxidant assay of selected aqueous plant extracts and their polyherbal formulation

    Directory of Open Access Journals (Sweden)

    Ganga Raju M.


    Full Text Available To support the use of selected plant extracts in Ayurveda, naturopathy, the antioxidant potential of the aqueous extract of Vincarosea (VR, Gymnemasylvestre (GS, Tinosporacordifolia (TC and Emblicaofficinalis (EO and their mixture (PHF of Indian origin was investigated for in vitro antioxidant activity by using in vitro models like superoxide, hydroxyl radical scavenging activity and lipid peroxide inhibition assay. The results were compared with standard (ascorbic acid, a known antioxidant. The various phytoconstituents identified in the above selected plants extracts were poly phenols, flavonoids, terpenoids, tannins, alkaloids. The terpenoids were reported to protect lipids, blood and body fluids against the attack of free radicals, some types of reactive oxygen, hydroxylic groups, peroxides and superoxide radicals. The presence of these phytoconstituents in selected plants might be responsible for antioxidant activity with that of known antioxidant ascorbic acid.

  12. Effects of several salt marsh plants on mouse spleen and thymus cell proliferation using mtt assay (United States)

    Seo, Youngwan; Lee, Hee-Jung; Kim, You Ah; Youn, Hyun Joo; Lee, Burm-Jong


    In the present study, we have tested the effects of 21 salt marsh plants on cell proliferation of mouse immune cells (spleen and thymus) using MTT assay in culture. The methanolic extracts of six salt marsh plants ( Rosa rugosa, Ixeris tamagawaensis, Artemisia capillaris, Tetragonia tetragonoides, Erigeron annus, and Glehnia littoralis) showed very powerful suppressive effects of mouse immune cell death and significant activities of cell proliferation in vitro. Especially, the methanolic extract of Rosa rugosa was found to have fifteen times compared to the control treatment, demonstrating that Rosa rugosa may have a potent stimulation effect on immune cell proliferation. These results suggest that several salt marsh plants including Rosa rugosa could be useful for further study as an immunomodulating agent.

  13. "Singing in the Tube"--audiovisual assay of plant oil repellent activity against mosquitoes (Culex pipiens). (United States)

    Adams, Temitope F; Wongchai, Chatchawal; Chaidee, Anchalee; Pfeiffer, Wolfgang


    Plant essential oils have been suggested as a promising alternative to the established mosquito repellent DEET (N,N-diethyl-meta-toluamide). Searching for an assay with generally available equipment, we designed a new audiovisual assay of repellent activity against mosquitoes "Singing in the Tube," testing single mosquitoes in Drosophila cultivation tubes. Statistics with regression analysis should compensate for limitations of simple hardware. The assay was established with female Culex pipiens mosquitoes in 60 experiments, 120-h audio recording, and 2580 estimations of the distance between mosquito sitting position and the chemical. Correlations between parameters of sitting position, flight activity pattern, and flight tone spectrum were analyzed. Regression analysis of psycho-acoustic data of audio files (dB[A]) used a squared and modified sinus function determining wing beat frequency WBF ± SD (357 ± 47 Hz). Application of logistic regression defined the repelling velocity constant. The repelling velocity constant showed a decreasing order of efficiency of plant essential oils: rosemary (Rosmarinus officinalis), eucalyptus (Eucalyptus globulus), lavender (Lavandula angustifolia), citronella (Cymbopogon nardus), tea tree (Melaleuca alternifolia), clove (Syzygium aromaticum), lemon (Citrus limon), patchouli (Pogostemon cablin), DEET, cedar wood (Cedrus atlantica). In conclusion, we suggest (1) disease vector control (e.g., impregnation of bed nets) by eight plant essential oils with repelling velocity superior to DEET, (2) simple mosquito repellency testing in Drosophila cultivation tubes, (3) automated approaches and room surveillance by generally available audio equipment (dB[A]: ISO standard 226), and (4) quantification of repellent activity by parameters of the audiovisual assay defined by correlation and regression analyses.

  14. Large-scale purification of porcine or bovine photoreceptor outer segments for phagocytosis assays on retinal pigment epithelial cells. (United States)

    Parinot, Célia; Rieu, Quentin; Chatagnon, Jonathan; Finnemann, Silvia C; Nandrot, Emeline F


    Analysis of one of the vital functions of retinal pigment epithelial (RPE) cells, the phagocytosis of spent aged distal fragments of photoreceptor outer segments (POS) can be performed in vitro. Photoreceptor outer segments with stacks of membranous discs containing the phototransduction machinery are continuously renewed in the retina. Spent POS are eliminated daily by RPE cells. Rodent, porcine/bovine and human RPE cells recognize POS from various species in a similar manner. To facilitate performing large series of experiments with little variability, a large stock of POS can be isolated from porcine eyes and stored frozen in aliquots. This protocol takes advantage of the characteristic of photopigments that display an orange color when kept in the dark. Under dim red light, retinae are collected in a buffer from opened eyecups cut in halves. The retinal cell suspension is homogenized, filtered and loaded onto a continuous sucrose gradient. After centrifugation, POS are located in a discrete band in the upper part of the gradient that has a characteristic orange color. POS are then collected, spun, resuspended sequentially in wash buffers, counted and aliquoted. POS obtained this way can be used for phagocytosis assays and analysis of protein activation, localization or interaction at various times after POS challenge. Alternatively, POS can be labeled with fluorophores, e.g., FITC, before aliquoting for subsequent fluorescence quantification of POS binding or engulfment. Other possible applications include the use of modified POS or POS challenge combined with stress conditions to study the effect of oxidative stress or aging on RPE cells.

  15. Good manufacturing practices production of a purification-free oral cholera vaccine expressed in transgenic rice plants. (United States)

    Kashima, Koji; Yuki, Yoshikazu; Mejima, Mio; Kurokawa, Shiho; Suzuki, Yuji; Minakawa, Satomi; Takeyama, Natsumi; Fukuyama, Yoshiko; Azegami, Tatsuhiko; Tanimoto, Takeshi; Kuroda, Masaharu; Tamura, Minoru; Gomi, Yasuyuki; Kiyono, Hiroshi


    The first Good Manufacturing Practices production of a purification-free rice-based oral cholera vaccine (MucoRice-CTB) from transgenic plants in a closed cultivation system yielded a product meeting regulatory requirements. Despite our knowledge of their advantages, plant-based vaccines remain unavailable for human use in both developing and industrialized countries. A leading, practical obstacle to their widespread use is producing plant-based vaccines that meet governmental regulatory requirements. Here, we report the first production according to current Good Manufacturing Practices of a rice-based vaccine, the cholera vaccine MucoRice-CTB, at an academic institution. To this end, we established specifications and methods for the master seed bank (MSB) of MucoRice-CTB, which was previously generated as a selection-marker-free line, evaluated its propagation, and given that the stored seeds must be renewed periodically. The production of MucoRice-CTB incorporated a closed hydroponic system for cultivating the transgenic plants, to minimize variations in expression and quality during vaccine manufacture. This type of molecular farming factory can be operated year-round, generating three harvests annually, and is cost- and production-effective. Rice was polished to a ratio of 95 % and then powdered to produce the MucoRice-CTB drug substance, and the identity, potency, and safety of the MucoRice-CTB product met pre-established release requirements. The formulation of MucoRice-CTB made by fine-powdering of drug substance and packaged in an aluminum pouch is being evaluated in a physician-initiated phase I study.

  16. Purification of a water extract of Chinese sweet tea plant (Rubus suavissimus S. Lee) by alcohol precipitation. (United States)

    Koh, Gar Yee; Chou, Guixin; Liu, Zhijun


    The aqueous extraction process of the leaves of Rubus suavissimus often brings in a large amount of nonactive polysaccharides as part of the constituents. To purify this water extract for potential elevated bioactivity, an alcohol precipitation (AP) consisting of gradient regimens was applied, and its resultants were examined through colorimetric and HPLC analyses. AP was effective in partitioning the aqueous crude extract into a soluble supernatant and an insoluble precipitant, and its effect varied significantly with alcohol regimens. Generally, the higher the alcohol concentration, the purer was the resultant extract. At its maximum, approximately 36% (w/w) of the crude extract, of which 23% was polysaccharides, was precipitated and removed, resulting in a purified extract consisting of over 20% bioactive marker compounds (gallic acid, ellagic acid, rutin, rubusoside, and steviol monoside). The removal of 11% polysaccharides from the crude water extract by using alcohol precipitation was complete at 70% alcohol regimen. Higher alcohol levels resulted in even purer extracts, possibly by removing some compounds of uncertain bioactivity. Alcohol precipitation is an effective way of removing polysaccharides from the water extract of the sweet tea plant and could be used as an initial simple purification tool for many water plant extracts that contain large amounts of polysaccharides.

  17. Methylated-antibody affinity purification to improve proteomic identification of plant RNA polymerase Pol V complex and the interacting proteins (United States)

    Qin, Guochen; Ma, Jun; Chen, Xiaomei; Chu, Zhaoqing; She, Yi-Min


    Affinity purification followed by enzymatic digestion and mass spectrometry has been widely utilized for the sensitive detection of interacting proteins and protein complexes in various organisms. In plants, the method is technically challenging due to the low abundance proteins, non-specific binding and difficulties of eluting interacting proteins from antibody beads. In this report, we describe a strategy to modify antibodies by reductive methylation of lysines without affecting their binding properties, followed by on-bead digestion of bound proteins with endoproteinase Lys-C. By this method, the antibody remains intact and does not interfere with the downstream identification of interacting proteins. Non-specific binding proteins were excluded using 14N/15N-metabolic labeling of wild-type and the transgenic plant counterparts. The method was employed to identify 12 co-immunoprecipitated protein subunits in Pol V complex and to discover 17 potential interacting protein targets in Arabidopsis. Our results demonstrated that the modification of antibodies by reductive dimethylation can improve the reliability and sensitivity of identifying low-abundance proteins through on-bead digestion and mass spectrometry. We also show that coupling this technique with chemical crosslinking enables in-depth characterization of endogenous protein complexes and the protein-protein interaction networks including mapping the surface topology and post-translational modifications of interacting proteins. PMID:28224978

  18. [Various Salmonella serotypes isolated at a sewage purification plant in a smaller city over a one-year period]. (United States)

    Schüsseler, G; Sobotta, B; Gerhardt, G G; Teitge, E; Gundermann, K O


    A one-year-study was carried out in the waste-water treatment plant of Plön (population equivalents 60,000), which has a mechanical and a biological purification and an additional chemical flocculation. Samples were taken at five different places in the plant and examined for Salmonella by use of membrane-filtration and MPN-method. 2,611 Salmonella-strains, representing 23 species, were isolated and serologically typed from samples taken at ten days. S. typhi-murium was found most frequently (Table 1, Fig. 1). The largest spectrum of different types was located in the activated sludge-basin and at the outlet of the chemical flocculation (Table 2). No correlation could be established between the qualitative findings and the Salmonella-counts or other parameter like temperatures. All the ten species that have been officially reported to cause salmonellosis in man were also isolated from the sewage (Table 5). Findings of other Salmonella-serotypes are attributed to unreported human infections and animals or other sources.

  19. Separation and purification of glucosinolates from crude plant homogenates by high-speed counter-current chromatography. (United States)

    Fahey, Jed W; Wade, Kristina L; Stephenson, Katherine K; Chou, F Edward


    Glucosinolates are anionic, hydrophilic plant secondary metabolites which are of particular interest due to their role in the prevention of cancer and other chronic and degenerative diseases. The separation and purification of glucosinolates from a variety of plant sources (e.g. seeds of broccoli, arugula and the horseradish tree), was achieved using high-speed counter-current chromatography (HSCCC). A high-salt, highly polar system containing 1-propanol-acetonitrile-saturated aqueous ammonium sulfate-water (1:0.5:1.2:1), was run on a semi-preparative scale and then transferred directly to preparative scale. Up to 7 g of a concentrated methanolic syrup containing about 10% glucosinolates was loaded on an 850-ml HSCCC column, and good separation and recovery were demonstrated for 4-methylsulfinylbutyl, 3-methylsulfinylpropyl, 4-methylthiobutyl, 2-propenyl and 4-(rhamnopyranosyloxy)benzyl glucosinolates. Multiple injections (5 to 6 times) were performed with well-preserved liquid stationary phase under centrifugal force. Pooled sequential runs with broccoli seed extract yielded about 20 g of its predominant glucosinolate, glucoraphanin, which was produced at > 95% purity and reduced to powdered form.

  20. A co-beneficial system using aquatic plants: bioethanol production from free-floating aquatic plants used for water purification. (United States)

    Soda, S; Mishima, D; Inoue, D; Ike, M


    A co-beneficial system using constructed wetlands (CWs) planted with aquatic plants is proposed for bioethanol production and nutrient removal from wastewater. The potential for bioethanol production from aquatic plant biomass was experimentally evaluated. Water hyacinth and water lettuce were selected because of their high growth rates and easy harvestability attributable to their free-floating vegetation form. The alkaline/oxidative pretreatment was selected for improving enzymatic hydrolysis of the aquatic plants. Ethanol was produced with yields of 0.14-0.17 g-ethanol/ g-biomass in a simultaneous saccharification and fermentation mode using a recombinant Escherichia coli strain or a typical yeast strain Saccharomyces cerevisiae. Subsequently, the combined benefits of the CWs planted with the aquatic plants for bioethanol production and nutrient removal were theoretically estimated. For treating domestic wastewater at 1,100 m(3)/d, it was inferred that the anoxic-oxic activated sludge process consumes energy at 3,200 MJ/d, whereas the conventional activated sludge process followed by the CW consumes only 1,800 MJ/d with ethanol production at 115 MJ/d.

  1. House-plant placement for indoor air purification and health benefits on asthmatics



    Objectives Some plants were placed in indoor locations frequented by asthmatics in order to evaluate the quality of indoor air and examine the health benefits to asthmatics. Methods The present study classified the participants into two groups: households of continuation and households of withdrawal by a quasi-experimental design. The households of continuation spent the two observation terms with indoor plants, whereas the households of withdrawal passed the former observation terms with ind...

  2. 室内植物对甲醛净化性能的研究进展%Progress of Research on the Purification of Formaldehyde by Plants

    Institute of Scientific and Technical Information of China (English)

    蔡宝珍; 金荷仙; 熊伟


    随着人们生活水平的提高,室内空气净化已成为人们普遍关心的问题,而甲醛作为室内空气污染的头号杀手,引起了人们的巨大关注.笔者简述室内甲醛污染的来源、危害以及净化甲醛的重要性.与物理吸附、化学吸收、光催化和生物降解等技术相比,植物净化甲醛技术具有自然、环保、操作简单的特点,已经成为室内甲醛净化和室内植物景观研究的热点和前沿科学之一.综述植物净化甲醛的国内外研究进展,着重从吸附净化、代谢净化及根际微生物的降解净化等3方面阐述植物净化甲醛的机理,介绍植物净化甲醛的毒理研究进展,同时指出目前研究中存在的主要问题,并进一步提出未来的研究发展方向.%Purification of indoor polluted air becomes a common concern topic along with living standards'enhancement.As the top killer of indoor polluted air, the formaldehyde pollution was of special concern.The origin of formaldehyde and its enormous pollution in indoor air were inevitable, so it was very import to purify indoor air formaldehyde.Compared with technology of physical absorption, chemical absorption, photocatalytic biodegradable, the purification of formaldehyde by plant was natural, environmental and easy operate, which became the hot topic and advanced science in the research of purification of indoor formaldehyde and houseplant landscape.The domestic and abroad research progress of aerobic granular sludge was summerized.The theory of purification of formaldehyde was the absorption by the stem and foliage, the metabolism and transformation by the plant, and the degradation by the rhizosphere microorganism.Toxicological purification of formaldehyde by indoor plants was viewed.Advantages and disadvantages in research were reported, too.Finally, the foreground of the purification of formaldehyde by indoor plants was prospected.

  3. Identification and assay of the flavonoids in medicinal plants with hepatoprotective action

    Directory of Open Access Journals (Sweden)

    Maria A. Cojocaru-Toma


    Full Text Available The article describes the identification and assay of the flavonoids in medicinal plants with hepatoprotective action, harvested as a culture at the Cultivation Center of the Medicinal Plants within State University of Medicine and Pharmacy „Nicolae Testemițanu” from the Republic of Moldova, using Pharmacopoeia methods. The flavonoids, found in the examined medicinal product, are responsible for hepatoprotective activity due to antioxidant activity, exhibited by neutralizing free radicals. The flavonoids were identified by using the Chinode method and thin layer chromatography, operating with 3 systems of solvents, and as biomarkers rutine, quercetine and luteolin ewere used. The results of the quantitative analysis point out that the content of the flavonoids determined by using spectrophotometric method is in the range of 0,620% to 1,204%, in studied vegetal products.

  4. A nylon membrane bag assay for determination of the effect of chemicals on soilborne plant pathogens in soil (United States)

    A new nylon membrane bag assay was developed that can rapidly and effectively determine the impact of chemicals added to soil on soilborne plant pathogens for which there are no selective media or for which a selective medium is expensive or difficult to prepare. This assay consists of placing patho...

  5. 天然气净化厂火炬系统设计研究%Design of the Flare System of Natural Gas Purification Plants

    Institute of Scientific and Technical Information of China (English)



    The flare system design of natural gas purification plants is different from ordinary refineries. The flare gas in natural gas purification plants has high sulfur content and low temperature. And current purification plants constructed by Sinopec Group require device maintenance in batches, namely the flare system can not shut down, which will propose new design issues and requirements.%天然气净化厂火炬系统的设计与普通炼油厂火炬系统设计有所不同。天然气净化厂中的放空火炬气含硫量高、放空温度低。而且目前中石化所建设的净化厂项目均要求装置轮番检修,即火炬系统不能停工,对设计提出了新的课题和要求。

  6. Plant life forms in thermal regulation and self purification of urban housing environments

    Energy Technology Data Exchange (ETDEWEB)

    Raza, S.H. [Osmania Univ., Ecology and Environmental Studies Div., Hyderabad (India)


    This article is concerned with the air quality of the closed indoor environment with respect to its temperature and carbon dioxide levels and with the assessment of management practices that have been used to reduce temperature and carbon dioxide levels with the help of certain plants. Phanerophytic lifeforms, such as trees, shrubs, herbs and creepers surrounding dwellings can be shown to produce a cooling effect, reducing temperatures by up to 11{sup o}C. Certain succulent plants like Kalanchoe marmorata, Bryophyllum pinnata and Apicra deltoideae were observed under experimental conditions to reduce carbon dioxide levels up to 90% from closed chambers under dark conditions. Certain ornamental plants such as Verbena bipinnatifida and Ixora coccinea could remove 63-75% of carbon dioxide from closed indoor environments in the presence of light. (author) 3 tabs., 11 refs.

  7. A comparative study of the radiological hazard in sediments samples from drinking water purification plants supplied from different sources

    Directory of Open Access Journals (Sweden)

    Shams A.M. Issa


    Full Text Available The natural radiation level has been determined for 135 sediment samples from forty-six drinking water purification plants supplied from different sources (Nile River, Ibrahimia Canal and Bahr Yousif Canal aiming to evaluate the radiation hazard. The concentration of natural radionuclides (226Ra, 232Th and 40K has been investigated by using gamma spectrometry (NaI (Tl 3″ × 3″ detector. The results showed that the concentrations of average activity in the sediment samples collected from Nile River, Ibrahimia Canal and Bahr Yousif Canal are (29 ± 2, 30 ± 2 and 240 ± 8 Bq kg−1, (47 ± 3, 46 ± 8 and 258 ± 12 Bq kg−1 and (28 ± 2, 27 ± 3 and 219 ± 18 Bq kg−1 for 226Ra, 232Th and 40K, respectively. The distributions of average activity concentrations of samples under investigation are within the world values although some extreme values have been determined. Radiological hazard effects such as: absorbed dose rate (D, outdoor and indoor annual effective dose equivalent (AEDE, radium equivalent activities (Raeq, hazard indices (Hex and Hin, gamma index (Iγ, excess lifetime cancer risk (ELCR and annual gonadal dose equivalent (AGDE for the corresponding samples were also estimated.

  8. Toxicity of selected plant volatiles in microbial and mammalian short-term assays. (United States)

    Stammati, A; Bonsi, P; Zucco, F; Moezelaar, R; Alakomi, H L; von Wright, A


    In this study, several short-term microbial and mammalian in vitro assays were used to evaluate cytotoxicity and genotoxicity of four plant volatiles showing antifungal activity: cinnamaldehyde, carvacrol, thymol and S(+)-carvone. All inhibited viability and proliferation of Hep-2 cells in a dose-dependent manner. IC50 ranged from 0.3 mM (cinnamaldehyde) to 0.7 mM (thymol) in viability tests and from 0.2 mM (carvacrol) to 0.9 mM (carvone) in the proliferation test. The morphological analysis suggested an involvement of apoptosis in the cases of carvone, carvacrol and cinnamaldehyde. At nontoxic doses, carvacrol and thymol increased the number of revertants in the Ames test by 1.5-1.7 times, regardless of metabolic activation. In the SOS-chromotest, none of the four plant volatiles caused DNA damage at non-toxic doses. In the DNA repair test, a marked dose-dependent differential toxicity was observed with carvone and, to a lesser extent, with cinnamaldehyde, while with thymol and carvacrol, this effect was less pronounced. In conclusion, the considered in vitro cytotoxicity assays have shown to be sensitive enough to highlight a variety of toxic effects at the cellular level, which can be rather different between chemically closely related compounds, such as isomers.

  9. Expression, purification and crystallization of a plant polyketide cyclase from Cannabis sativa. (United States)

    Yang, Xinmei; Matsui, Takashi; Mori, Takahiro; Taura, Futoshi; Noguchi, Hiroshi; Abe, Ikuro; Morita, Hiroyuki


    Plant polyketides are a structurally diverse family of natural products. In the biosynthesis of plant polyketides, the construction of the carbocyclic scaffold is a key step in diversifying the polyketide structure. Olivetolic acid cyclase (OAC) from Cannabis sativa L. is the only known plant polyketide cyclase that catalyzes the C2-C7 intramolecular aldol cyclization of linear pentyl tetra-β-ketide-CoA to generate olivetolic acid in the biosynthesis of cannabinoids. The enzyme is also thought to belong to the dimeric α+β barrel (DABB) protein family. However, because of a lack of functional analysis of other plant DABB proteins and low sequence identity with the functionally distinct bacterial DABB proteins, the catalytic mechanism of OAC has remained unclear. To clarify the intimate catalytic mechanism of OAC, the enzyme was overexpressed in Escherichia coli and crystallized using the vapour-diffusion method. The crystals diffracted X-rays to 1.40 Å resolution and belonged to space group P3121 or P3221, with unit-cell parameters a = b = 47.3, c = 176.0 Å. Further crystallographic analysis will provide valuable insights into the structure-function relationship and catalytic mechanism of OAC.

  10. Purification ability and carbon dioxide flux from surface flow constructed wetlands treating sewage treatment plant effluent. (United States)

    Wu, Haiming; Lin, Li; Zhang, Jian; Guo, Wenshan; Liang, Shuang; Liu, Hai


    In this study, a two-year experiment was carried out to investigate variation of carbon dioxide (CO2) flux from free water surface constructed wetlands (FWS CW) systems treating sewage treatment plant effluent, and treatment performance was also evaluated. The better 74.6-76.6% COD, 92.7-94.4% NH4(+)-N, 60.1-84.7% TN and 49.3-70.7% TP removal efficiencies were achieved in planted CW systems compared with unplanted systems. The planted CW was a net CO2 sink, while the unplanted CW was a net CO2 source in the entire study period. An obvious annual and seasonal variability of CO2 fluxes from different wetland systems was also presented with the average CO2 flux ranging from -592.83mgm(-2)h(-1) to 553.91mgm(-2)h(-1) during 2012-2013. In addition, the net exchange of CO2 between CW systems and the atmosphere was significantly affected by air temperature, and the presence of plants also had the significant effect on total CO2 emissions. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Efficient ASK-assisted system for expression and purification of plant F-box proteins. (United States)

    Li, Haiou; Yao, Ruifeng; Ma, Sui; Hu, Shuai; Li, Suhua; Wang, Yupei; Yan, Chun; Xie, Daoxin; Yan, Jianbin


    Ubiquitin-mediated protein degradation plays an essential role in plant growth and development as well as responses to environmental and endogenous signals. F-box protein is one of the key components of the SCF (SKP1-CUL1-F-box protein) E3 ubiquitin ligase complex, which recruit specific substrate proteins for subsequent ubiquitination and 26S proteasome-mediated degradation to regulate developmental processes and signaling networks. However, it is not easy to obtain purified F-box proteins with high activity due to their unstable protein structures. Here, we found that Arabidopsis SKP-like proteins (ASKs) can significantly improve soluble expression of F-box proteins and maintain their bioactivity. We established an efficient ASK-assisted method to express and purify plant F-box proteins. The method meets a broad range of criteria required for the biochemical analysis or protein crystallization of plant F-box proteins. © 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd.

  12. Leaching and selective copper recovery from acidic leachates of Três Marias zinc plant (MG, Brazil) metallurgical purification residues. (United States)

    Sethurajan, Manivannan; Huguenot, David; Lens, Piet N L; Horn, Heinrich A; Figueiredo, Luiz H A; van Hullebusch, Eric D


    Zinc plant purification residue (ZPR), a typical Zn-hydrometallurgical waste, was collected from the Três Marias Zn plant (MG, Brazil). ZPR was characterized for its metal content and fractionation, mineralogy, toxicity and leachability. Toxicity characteristics leaching procedure (TCLP) and European Community Bureau of Reference (BCR) sequential extraction results revealed that this ZPR displays high percentages of metals (Cd, Cu, Zn and Pb) in the highly mobilizable fractions, increasing its hazardous potential. Bulk chemical analysis, pH dependent leaching and acid (H2SO4) leaching studies confirm that the ZPR is polymetallic, rich in Cd, Cu and Zn. The sulfuric acid concentration (1 M), agitation speed (450 rpm), temperature (40 °C) and pulp density (20 g L(-1)) were optimized to leach the maximum amount of heavy metals (Cd, Cu and Zn). Under optimum conditions, more than 50%, 70% and 60% of the total Cd, Cu and Zn present in the ZPR can be leached, respectively. The metals in the acid leachates were investigated for metal sulfide precipitation with an emphasis on selective Cu recovery. Metal sulfide precipitation process parameters such as initial pH and Cu to sulfide ratio were optimized as pH 1.5 and 1:0.5 (Cu:sulfide) mass ratio, respectively. Under optimum conditions, more than 95% of Cu can be selectively recovered from the polymetallic ZPR leachates. The Cu precipitates characterization studies reveal that they are approximately 0.1 μm in diameter and mainly consist of Cu and S. XRD analysis showed covellite (CuS), chalcanthite (CuSO4·5H2O) and natrochalcite (NaCu2(SO4)2(OH)·H2O) as the mineral phases. ZPRs can thus be considered as an alternative resource for copper production.

  13. Acetone enhances the direct analysis of total condensed tannins in plant tissues by the butanol-HCl-iron assay (United States)

    The butanol-HCl spectrophotometric assay is widely used to quantify extractable and insoluble forms of condensed tannin (CT, syn. proanthocyanidin) in foods, feeds, and foliage of herbaceous and woody plants. However, this method underestimates total CT content when applied directly to plant materia...

  14. Monoclonal Antibody-Based Enzyme Linked Immunosorbent Assay for the Analysis of Jasmonates in Plants

    Institute of Scientific and Technical Information of China (English)

    Aixing Deng; Weiming Tan; Suping He; Wei Liu; Tiegui Nan; Zhaohu Li; Baomin Wang; Qing X.Li


    Methyl jasmonate (MeJA) and its free-acid form,jasmonic acid (JA) are naturally occurring plant growth regulators widely distributed in higher plants.In order to improve the sensitivity for the analysis of MeJA at low levels in small amounts of plant samples,a monoclonal antibody (MAb) (designated as MAb 3E5D7C4B6) against MeJA was derived from a JAbovine serum albumin (BSA) conjugate as an immunogen.The antibody belongs to the IgG1 subclass with a κ type light chain and has a dissociation constant of approximately 6.07 x 10-9 M.MAb3E5D7C4B6 is very specific to MeJA.It was used to develop a direct competitive enzyme-linked immunosorbent assay (dcELISA),conventional and simplified indirect competitive ELISAs (icELISA).JA was derivatized into MeJA for the ELISA analysis.The IC50 value and detection range for MeJA were,respectively,34 and 4-257 ng/mL by the conventional icELISA,21 and 3-226 ng/mL by the simplified icELISA and 5.0 and 0.7-97.0 ng/mL by the dcELISA.The dcELISA was more sensitive than either the conventional or simplified icELISA.The assays were used to measure the content of jasmonates as MeJA in tobacco leaves under drought stress or inoculated with tobacco mosaic virus and tomato leaves inoculated with tomato mosaic virus or Lirioinyza sativae Blanchard as compared with the corresponding healthy leaves.The increased jasmonates content indicated its role in response to the drought stress and pathogens.

  15. Near-Zero Emissions Oxy-Combustion Flue Gas Purification - Power Plant Performance

    Energy Technology Data Exchange (ETDEWEB)

    Andrew Seltzer; Zhen Fan


    A technical feasibility assessment was performed for retrofitting oxy-fuel technology to an existing power plant burning low sulfur PRB fuel and high sulfur bituminous fuel. The focus of this study was on the boiler/power generation island of a subcritical steam cycle power plant. The power plant performance in air and oxy-firing modes was estimated and modifications required for oxy-firing capabilities were identified. A 460 MWe (gross) reference subcritical PC power plant was modeled. The reference air-fired plant has a boiler efficiency (PRB/Bituminous) of 86.7%/89.3% and a plant net efficiency of 35.8/36.7%. Net efficiency for oxy-fuel firing including ASU/CPU duty is 25.6%/26.6% (PRB/Bituminous). The oxy-fuel flue gas recirculation flow to the boiler is 68%/72% (PRB/bituminous) of the flue gas (average O{sub 2} in feed gas is 27.4%/26.4%v (PRB/bituminous)). Maximum increase in tube wall temperature is less than 10ºF for oxy-fuel firing. For oxy-fuel firing, ammonia injected to the SCR was shut-off and the FGD is applied to remove SOx from the recycled primary gas stream and a portion of the SOx from the secondary stream for the high sulfur bituminous coal. Based on CFD simulations it was determined that at the furnace outlet compared to air-firing, SO{sub 3}/SO{sub 2} mole ratio is about the same, NOx ppmv level is about the same for PRB-firing and 2.5 times for bituminous-firing due to shutting off the OFA, and CO mole fraction is approximately double. A conceptual level cost estimate was performed for the incremental equipment and installation cost of the oxyfuel retrofit in the boiler island and steam system. The cost of the retrofit is estimated to be approximately 81 M$ for PRB low sulfur fuel and 84 M$ for bituminous high sulfur fuel.

  16. [Various Salmonella serotypes isolated at a sewage purification plant in a large city over a one-year period]. (United States)

    Sobotta, B; Schüsseler, G; Gerhardt, G G; Teitge, E; Gundermann, K O


    The paper offers the results of a one-year-survey of Salmonella-serotypes in a municipal sewage-purification plant with a capacity of roughly 70,000 m3 per day. Findings of a quantitative study had shown Salmonella-maxima in the activated-sludge-basin. This suggested specialized Salmonella-serotypes, resident in this part of the plant as a possible explanation which was to be verified by this study. On ten days samples were taken from the inlet, and the outlet of the primary-sedimentation-tank, the outlet of the activated-sludge-basin and the effluent of the final sedimentation-basin. A combination of membrane-filtration and MPN-Method with a fifefold enrichment in 2.5% tetrathionate was applied for salmonella isolation. Plating was done on malachit-green-chinablue-lactose-agar followed by serological typing. 1,587 strains representing 38 different serotypes (Table 1) were identified with S. typhi-murium (Fig. 1) accounting for 36% of the isolations followed by S. bovis-morbificans, S. hadar (Fig. 2) and S. panama. None of the serotypes found showed a preference of a special sampling point. The qualitative and quantitative distribution of Salmonella in the plant seems to depend on the Salmonella contents of the entering waste water mainly. The greatest variety of Salmonella-serotypes was located in the activated-sludge-basin (Table 2) where oxygen-enrichment seems to result in the best ecological conditions for Salmonella survival. 3.3% of 722 strains examined did not produce hydrogen-sulphide (Table 3) and some showed damaged flagella-antigens. As a possible explanation toxic influences in the sewage are discussed. The epidemiological links between findings of Salmonella in sewage and in man of the same area are established and results differing in some aspects explained by the high rate of unknown infections. The existence of an autochthonous Salmonella-population in the sewage plant could not be proved.

  17. In vitro pharmacodynamic evaluation of antiviral medicinal plants using a vector-based assay technique. (United States)

    Esimone, C O; Grunwald, T; Wildner, O; Nchinda, G; Tippler, B; Proksch, P; Uberla, K


    Medicinal plants are increasingly being projected as suitable alternative sources of antiviral agents. The development of a suitable in vitro pharmacodynamic screening technique could contribute to rapid identification of potential bioactive plants and also to the standardization and/or pharmacokinetic-pharmacodynamic profiling of the bioactive components. Recombinant viral vectors (lentiviral, retroviral and adenoviral) transferring the firefly luciferase gene were constructed and the inhibition of viral vector infectivity by various concentrations of plant extracts was evaluated in HeLa or Hep2 cells by measuring the changes in luciferase activity. Cytotoxicity of the extracts was evaluated in parallel on HeLa or Hep2 cells stably expressing luciferase. Amongst the 15 extracts screened, only the methanol (ME) and the ethyl acetate (ET) fractions of the lichen, Ramalina farinacea specifically reduced lentiviral and adenoviral infectivity in a dose-dependent manner. Further, chromatographic fractionation of ET into four fractions (ET1-ET4) revealed only ET4 to be selectively antiviral with an IC50 in the 20 microg ml(-1) range. Preliminary mechanistic studies based on the addition of the extracts at different time points in the viral infection cycle (kinetic studies) revealed that the inhibitory activity was highest if extract and vectors were preincubated prior to infection, suggesting that early steps in the lentiviral or adenoviral replication cycle could be the major target of ET4. Inhibition of wild-type HIV-1 was also observed at a 10-fold lower concentration of the extract. The vector-based assay is a suitable in vitro pharmacodynamic evaluation technique for antiviral medicinal plants. The technique has successfully demonstrated the presence of antiviral principles in R. farinacea. Potential anti-HIV medicinal plants could rapidly be evaluated with the reported vector-based technique. The lichen, R. farinacea could represent a lead source of antiviral

  18. Improving the technology for sulfuric acid purification of BTK fraction at the Makeyevsk Coke Chemical Plant. Sovershenstvovaniye tekhnologii sernokislotnoy ochistki fraktsii BTK na Makeyevskom koksokhimzavode

    Energy Technology Data Exchange (ETDEWEB)

    Korobchanskiy, V.I.; Dobrovolskaya, L.Ye.; Dzekunov, S.N.; Grebennikova, S.S.; Korobchanskiy, Yu.V.


    Data are cited about a study of the operation of an installation for sulfuric acid purification of a BTK fraction at the Makeyevsk Coke Chemical Plant (KKhZ). The purification is performed in a radiator mixer with the addition of unsaturated compounds, a piperylene fraction. The capability is shown of producing benzene for nitration of a high quality in existing equipment with an unchanged expenditure of raw material and secondary materials. The following ways for improvement are recommended: reduce the final boiling point of the fraction to 125 to 130 degrees; feed the piperylene fraction in two stages: behind the mixer pump and in the central part of the radiator mixer; and reduce the process length to 6 to 8 minutes.

  19. 4种植物组合对室内甲醛气体净化能力的研究%Study on 4 Kinds of Combination of Plant PurificAtion Ability of Indoor Formaldehyde

    Institute of Scientific and Technical Information of China (English)

    闫晓煜; 许丽颖; 徐强; 杜兴臣


    甲醛是室内装修污染的重要污染物之一,选用4种室内常用植物,将植物进行排列组合后,跟踪检测室内甲醛浓度变化,分析出各种植物组合对室内甲醛净化能力由大到小依次为吊兰和龙舌兰>吊兰和虎尾兰>虎尾兰和龙舌兰>吊兰和常春藤>常春藤和龙舌兰>常春藤和虎尾兰。研究发现,将植物进行适当合理的搭配可达到较强的净化效果。%Formaldehyde is one of the major pollutants in indoor decoration pollution ,paper selects four kinds of com‐mon indoor plants ,the plants permutations and combinations ,the tracking and detection of changes in the concen‐tration of indoor formaldehyde ,analyze various combinations of plants in descending sequence of indoor formalde‐hyde purification capacity as spider plants and agave>spider and Sansevieria>Sansevieria and agave>spider and Ivy> Ivy and agave>ivy and Sansevieria .The study found that the plants can be achieved with appropriate and reason‐able strong purifying effect .

  20. Development of an enzyme-linked immunosorbent assay for the veratrum plant teratogens: cyclopamine and jervine. (United States)

    Lee, Stephen T; Panter, Kip E; Gaffield, William; Stegelmeier, Bryan L


    Veratrum californicum was responsible for large losses of sheep grazing high mountain ranges in central Idaho in the 1950s. Veratrum induces various birth defects including the cyclopic-type craniofacial defect (monkey-faced lambs) that is specifically induced in lambs after pregnant ewes grazed the plant on the 14th day of gestation. The steroidal alkaloids cyclopamine (1) and jervine (2) were isolated from Veratrum and shown to be primarily responsible for the malformations. Cyclopamine (1) and jervine (2) are potent teratogens that inhibit Sonic hedgehog (Shh) signaling during gastrulation-stage embryonic development, producing cyclopia and holoprosencephaly. Although losses to the sheep industry from Veratrum are now relatively infrequent, occasional incidents of toxicoses and craniofacial malformations are still reported in sheep and other species. However, the benefits to biomedical research using cyclopamine (1) as a tool to study human diseases have greatly expanded. A competitive inhibition enzyme-linked immunosorbent assay (ELISA) to detect and measure cyclopamine (1) and jervine (2) was developed using polyclonal antibodies produced in ewes. The limits of detection of the assay were 90.0 and 22.7 pg for cyclopamine (1) and jervine (2), respectively. This assay was used for the detection and measurement of cyclopamine (1) spiked into sheep blood. The simple extraction-ELISA methods developed in this study demonstrate the potential of using these techniques for the rapid screening of biological samples to detect the presence and concentration of cyclopamine (1) and jervine (2) and will be beneficial to pharmacological studies and livestock diagnostics.

  1. Antimycobacterial screening of traditional medicinal plants using the microplate resazurin assay. (United States)

    Webster, Duncan; Lee, Timothy D G; Moore, Jill; Manning, Tracy; Kunimoto, Dennis; LeBlanc, Darren; Johnson, John A; Gray, Christopher A


    Multidrug-resistant Mycobacterium tuberculosis strains have rapidly become a global health concern. North American First Nations communities have used traditional medicines for generations to treat many pulmonary infections. In this study, we evaluated the antimycobacterial activity of 5 medicinal plants traditionally used as general therapeutics for pulmonary illnesses and specifically as treatments for tuberculosis. Aqueous extracts of Aralia nudicaulis, Symplocarpus foetidus, Heracleum maximum, Juniperus communis, and Acorus calamus were screened for antimycobacterial activity against Bacillus Calmette-Guérin, Mycobacterium avium, and M. tuberculosis H37Ra using the colorimetric microplate resazurin assay. Extracts of Acorus calamus and H. maximum root demonstrated significant antimycobacterial activity comparable to that of the rifampin control (2 microg/mL). Evaluation of the cytotoxicity of these 2 extracts using the MTT assay also showed that the extracts were less toxic to 3 human cell lines than was the DMSO positive control. This study demonstrates that aqueous extracts of the roots of H. maximum and Acorus calamus possess strong in vitro antimycobacterial activity, validates traditional knowledge, and provides potential for the development of urgently needed novel antituberculous therapeutics.

  2. Evaluation of Effectiveness Technological Process of Water Purification Exemplified on Modernized Water Treatment Plant at Otoczna

    Directory of Open Access Journals (Sweden)

    Jordanowska Joanna


    Full Text Available The article presents the work of the Water Treatment Plant in the town of Otoczna, located in the Wielkopolska province, before and after the modernization of the technological line. It includes the quality characteristics of the raw water and treated water with particular emphasis on changes in the quality indicators in the period 2002 -2012 in relation to the physicochemical parameters: the content of total iron and total manganese, the ammonium ion as well as organoleptic parameters(colour and turbidity. The efficiency of technological processes was analysed, including the processes of bed start up with chalcedonic sand to remove total iron and manganese and ammonium ion. Based on the survey, it was found that the applied modernization helped solve the problem of water quality, especially the removal of excessive concentrations of iron, manganese and ammonium nitrogen from groundwater.

  3. Purification of oily wastewater in thermal power plant by coal fly ash

    Institute of Scientific and Technical Information of China (English)

    刘仁龙; 陈维; 刘清才


    The removal of oil from oily wastewater in thermal power plant by coal fly ash(CFA) was investigated. It contained about 2.5 g/L of mineral oil,which had to be treated efficiently before it was discharged. The experiments were carried out as a function of different initial concentrations of oil,mass dosage,contact time and pH value to obtain the optimum conditions for the removal of oil from oily wastewater. The experimental results show that CFA presents the most suitable conditions for the removal of oil from waste-water at a dosage of CFA 2.5 g/L,15 min of contact time and a pH value of 4.8. The adsorption process is performed with almost 96% of oil removal from wastewater. The kinetic data meet the second-order kinetic model. The Langmuir and Freundlich adsorption models were applied to describing the experimental isotherms and isotherm constants. The equilibrium data fit very well with the Freundlich model.

  4. Photocatalysis for continuous air purification in wastewater treatment plants: from lab to reality. (United States)

    Portela, R; Tessinari, R F; Suárez, S; Rasmussen, S B; Hernández-Alonso, M D; Canela, M C; Avila, P; Sánchez, B


    The photocatalytic efficiency of TiO(2)-SiMgO(x) plates to oxidize H(2)S was first evaluated in a flat laboratory reactor with 50 mL min(-1) synthetic air containing 100 ppm H(2)S in the presence of humidity. The use of the photocatalyst-adsorbent hybrid material enhanced the photocatalytic activity in terms of pollutant conversion, selectivity, and catalyst lifetime compared to previous H(2)S tests with pure TiO(2) because total H(2)S elimination was maintained for more than 30 operating hours with SO(2) appearing in the outlet as reaction product only after 18 h. Subsequently, the hybrid material was successfully tested in a photoreactor prototype to treat real polluted air in a wastewater treatment plant. For this purpose, a new tubular photocatalytic reactor that may use solar radiation in combination with artificial radiation was designed; the lamp was turned on when solar UV-A irradiance was below 20 W m(-2), which was observed to be the minimum value to ensure 100% conversion. The efficient distribution of the opaque photocatalyst inside the tubular reactor was achieved by using especially designed star-shaped structures. These structures were employed for the arrangement of groups of eight TiO(2)-SiMgO(x) plates in easy-to-handle channelled units obtaining an adequate flow regime without shading. The prototype continuously removed during one month and under real conditions the H(2)S contained in a 1 L min(-1) air current with a variable inlet concentration in the range of tens of ppmv without release of SO(2).

  5. The compost with the OFMSW as a solution for the sludges from the wastewater purification plants; Compostaje con FORSU como una solucion para los fangos de depuradora

    Energy Technology Data Exchange (ETDEWEB)

    Chica Perez, A.; Diaz Rubio, M. M.; Mohedo Gaton, J. J.; Martin Martin, A. [Universidad de Cardoba (Spain); Revilla Alvarez, J. R.


    The correct management of the sludges coming from the municipal wastewater purification plants (mwpp) is an unsolved problem in many cases and its volume is quickly increasing. As a solution for its treatment and disposal the composting process of the sludges combined with the organic fraction of municipal solid wastes (OFMSW) selectively collected is proposed in this work. A suitable compost for agricultural use is obtained. Using respirometer methods better than using other analytical parameters can monitor the stabilisation process of the mixture of sludges from MWPP and OFMSW. The measurement of the maximum oxygen intake specific rate indicates an appropriate stabilisation of the mixture after third treatment month. (Author) 32 refs.

  6. Penicillinase-based enzyme-linked immunosorbent assay for the detection of plant viruses. (United States)

    Sudarshana, M R; Reddy, D V


    A penicillinase (PNC)-based, enzyme-linked immunosorbent assay (ELISA) was standardized to detect maize mosaic virus (MMV) in sorghum leaf extracts, peanut mottle virus (PMV) in pea leaf extracts, and tomato spotted wilt virus (TSWV) in peanut leaf extracts. Rabbit Fc-specific antibodies were conjugated with PNC by a single step glutaraldehyde bridge. Among several indicators tested, bromothymol blue (BTB) was found suitable for measuring PNC activity under simulated conditions. Two reagents, starch-iodine complex (SIC) and a mixed pH indicator, containing bromocresol purple and BTB (2:1) used earlier for the PNC-based ELISA, were compared with BTB for utilization in the PNC-based ELISA. SIC gave a slightly higher virus titre than BTB or the mixed pH indicator, but it often gave nonspecific reactions. Sodium or potassium salts of penicillin-G at 0.5-1.0 mg/ml and BTB at 0.2 mg/ml were found to be suitable as substrate-indicator mixture for PNC-based ELISA. The sensitivity of the PNC system was comparable to those of the alkaline phosphatase (ALP) and horseradish peroxidase (HRP) systems in detecting MMV, PMV, and TSWV. The PNC conjugate could be used at a greater dilution than those of the ALP and HRP conjugates and the BTB substrate mixture was stable for at least 3 weeks at 4 degrees C. Penicillin is readily available in developing countries, and at a substantially lower cost than p-nitrophenyl phosphate, the commonly used substrate for ALP in the plate ELISA. Thus the PNC-based ELISA provides a less expensive means for assaying plant viruses by ELISA.

  7. Choosing the best plant for the job: a cost-effective assay to prescreen ancient plant remains destined for shotgun sequencing.

    Directory of Open Access Journals (Sweden)

    Nathan Wales

    Full Text Available DNA extracted from ancient plant remains almost always contains a mixture of endogenous (that is, derived from the plant and exogenous (derived from other sources DNA. The exogenous 'contaminant' DNA, chiefly derived from microorganisms, presents significant problems for shotgun sequencing. In some samples, more than 90% of the recovered sequences are exogenous, providing limited data relevant to the sample. However, other samples have far less contamination and subsequently yield much more useful data via shotgun sequencing. Given the investment required for high-throughput sequencing, whenever multiple samples are available, it is most economical to sequence the least contaminated sample. We present an assay based on quantitative real-time PCR which estimates the relative amounts of fungal and bacterial DNA in a sample in comparison to the endogenous plant DNA. Given a collection of contextually-similar ancient plant samples, this low cost assay aids in selecting the best sample for shotgun sequencing.

  8. Tuning a 96-Well Microtiter Plate Fluorescence-Based Assay to Identify AGE Inhibitors in Crude Plant Extracts

    Directory of Open Access Journals (Sweden)

    Luc Séro


    Full Text Available Advanced glycation end-products (AGEs are involved in the pathogenesis of numerous diseases. Among them, cellular accumulation of AGEs contributes to vascular complications in diabetes. Besides using drugs to lower blood sugar, a balanced diet and the intake of herbal products potentially limiting AGE formation could be considered beneficial for patients’ health. The current paper presents a simple and cheap high-throughput screening (HTS assay based on AGE fluorescence and suitable for plant extract screening. We have already implemented an HTS assay based on vesperlysines-like fluorescing AGEs quickly (24 h formed from BSA and ribose under physiological conditions. However, interference was noted when fluorescent compounds and/or complex mixtures were tested. To overcome these problems and apply this HTS assay to plant extracts, we developed a technique for systematic quantification of both vesperlysines (λexc 370 nm; λem 440 nm and pentosidine-like (λexc 335 nm; λem 385 nm AGEs. In a batch of medicinal and food plant extracts, hits were selected as soon as fluorescence decreased under a fixed threshold for at least one wavelength. Hits revealed during this study appeared to contain well-known and powerful anti-AGE substances, thus demonstrating the suitability of this assay for screening crude extracts (0.1 mg/mL. Finally, quercetin was found to be a more powerful reference compound than aminoguanidine in such assay.

  9. A Simple and Reliable Assay for Detecting Specific Nucleotide Sequences in Plants Using Optical Thin-film Biosensor Chips

    Institute of Scientific and Technical Information of China (English)

    S. Bai; X. Zhong; L. Ma; W. Zheng; L. Fan; N. Wei; X.W. Deng


    @@ Here we report the adaptation and optimization of an efficient, accurate and inexpensive assay that employs custom-designed silicon-based optical thin-film biosensor chips to detect unique transgenes in genetically modified (GM) crops and SNP markers in model plant genomes.

  10. Tier-1 assays for assessing the toxicity of insecticidal proteins produced by genetically engineered plants to non-target arthropods. (United States)

    Li, Yun-He; Romeis, Jörg; Wu, Kong-Ming; Peng, Yu-Fa


    In assessing an insect-resistant genetically engineered (IRGE) crop before its commercialization, researchers normally use so-called "Tier-1 assays" as the initial step to determine the effects of the crop on non-target organisms. In these tests, the insecticidal proteins (IPs) produced by the IRGEs are added to the diets of test organisms in the laboratory. Test organisms in such assays can be directly exposed to much higher concentrations of the test IPs than they would encounter in the field. The results of Tier-1 assays are thus more conservative than those generated in studies in which the organisms are exposed to the IPs by feeding on IRGE plant tissue or in the case of predators or parasites, by feeding on invertebrate prey or hosts that have fed on IRGE plant tissue. In this report, we consider three important factors that must be considered in Tier-1 assays: (i) methods for delivery of the IP to the test organisms; (ii) the need for and selection of compounds used as positive controls; and (iii) methods for monitoring the concentration, stability and bioactivity of the IP during the assay. We also analyze the existing data from Tier-1 assays regarding the toxicity of Bt Cry proteins to non-target arthropod species. The data indicate that the widely used Bt proteins have no direct toxicity to non-target organisms.

  11. An optimized microplate assay system for quantitative evaluation of plant cell wall-degrading enzyme activity of fungal culture extracts. (United States)

    King, Brian C; Donnelly, Marie K; Bergstrom, Gary C; Walker, Larry P; Gibson, Donna M


    Developing enzyme cocktails for cellulosic biomass hydrolysis complementary to current cellulase systems is a critical step needed for economically viable biofuels production. Recent genomic analysis indicates that some plant pathogenic fungi are likely a largely untapped resource in which to prospect for novel hydrolytic enzymes for biomass conversion. In order to develop high throughput screening assays for enzyme bioprospecting, a standardized microplate assay was developed for rapid analysis of polysaccharide hydrolysis by fungal extracts, incorporating biomass substrates. Fungi were grown for 10 days on cellulose- or switchgrass-containing media to produce enzyme extracts for analysis. Reducing sugar released from filter paper, Avicel, corn stalk, switchgrass, carboxymethylcellulose, and arabinoxylan was quantified using a miniaturized colorimetric assay based on 3,5-dinitrosalicylic acid. Significant interactions were identified among fungal species, growth media composition, assay substrate, and temperature. Within a small sampling of plant pathogenic fungi, some extracts had crude activities comparable to or greater than T. reesei, particularly when assayed at lower temperatures and on biomass substrates. This microplate assay system should prove useful for high-throughput bioprospecting for new sources of novel enzymes for biofuel production.

  12. A cytogenetic study of nuclear power plant workers using the micronucleus-centromere assay. (United States)

    Thierens, H; Vral, A; Barbé, M; Aousalah, B; De Ridder, L


    A cytogenetic study was performed in 215 nuclear power plant workers occupationally exposed to radiation using the micronucleus-centromere assay for peripheral blood lymphocytes. As control population served administrative staff with yearly doses below 1 mSv. The increase of the micronucleus frequency with age, observed in the non-smoking control population, is mainly due to an enhanced number of centromere-positive micronuclei, pointing to an increased chromosome loss. No differences in the number of micronuclei, centromere-positive and centromere-negative micronuclei between smokers and non-smokers are observed. An analysis of the micronucleus data vs. the dose accumulated over the 10 years preceding the venepuncture shows no significant clastogenic or aneuploidogenic effects of the exposure in the studied population which is representative for workers in the nuclear industry at present. According to the linear fits to our data an increase of the micronucleus frequency pro rata 0.5 per 1000 binucleated cells per year, related to the centromere-negative micronuclei, may be expected for workers with the maximal tolerable dose of 20 mSv/year.

  13. Teaching about citric acid cycle using plant mitochondrial preparations: Some assays for use in laboratory courses*. (United States)

    Vicente, Joaquim A F; Gomes-Santos, Carina S S; Sousa, Ana Paula M; Madeira, Vítor M C


    Potato tubers and turnip roots were used to prepare purified mitochondria for laboratory practical work in the teaching of the citric acid cycle (TCA cycle). Plant mitochondria are particularly advantageous over the animal fractions to demonstrate the TCA cycle enzymatic steps, by using simple techniques to measure O(2) consumption and transmembrane potential (ΔΨ). The several TCA cycle intermediates induce specific enzyme activities, which can be identified by respiratory parameters. Such a strategy is also used to evidence properties of the TCA cycle enzymes: ADP stimulation of isocitrate dehydrogenase and α-ketoglutarate dehydrogenase; activation by citrate of downstream oxidation steps, e.g. succinate dehydrogenase; and regulation of the activity of isocitrate dehydrogenase by citrate action on the citrate/isocitrate carrier. Furthermore, it has been demonstrated that, in the absence of exogenous Mg(2+) , isocitrate-dependent respiration favors the alternative oxidase pathway, as judged by changes of the ADP/O elicited by the inhibitor n-propyl galate. These are some examples of assays related with TCA cycle intermediates we can use in laboratory courses. Copyright © 2005 International Union of Biochemistry and Molecular Biology, Inc.

  14. Can physiological endpoints improve the sensitivity of assays with plants in the risk assessment of contaminated soils?

    Directory of Open Access Journals (Sweden)

    Ana Gavina

    Full Text Available Site-specific risk assessment of contaminated areas indicates prior areas for intervention, and provides helpful information for risk managers. This study was conducted in the Ervedosa mine area (Bragança, Portugal, where both underground and open pit exploration of tin and arsenic minerals were performed for about one century (1857-1969. We aimed at obtaining ecotoxicological information with terrestrial and aquatic plant species to integrate in the risk assessment of this mine area. Further we also intended to evaluate if the assessment of other parameters, in standard assays with terrestrial plants, can improve the identification of phytotoxic soils. For this purpose, soil samples were collected on 16 sampling sites distributed along four transects, defined within the mine area, and in one reference site. General soil physical and chemical parameters, total and extractable metal contents were analyzed. Assays were performed for soil elutriates and for the whole soil matrix following standard guidelines for growth inhibition assay with Lemna minor and emergence and seedling growth assay with Zea mays. At the end of the Z. mays assay, relative water content, membrane permeability, leaf area, content of photosynthetic pigments (chlorophylls and carotenoids, malondialdehyde levels, proline content, and chlorophyll fluorescence (Fv/Fm and ΦPSII parameters were evaluated. In general, the soils near the exploration area revealed high levels of Al, Mn, Fe and Cu. Almost all the soils from transepts C, D and F presented total concentrations of arsenic well above soils screening benchmark values available. Elutriates of several soils from sampling sites near the exploration and ore treatment areas were toxic to L. minor, suggesting that the retention function of these soils was seriously compromised. In Z. mays assay, plant performance parameters (other than those recommended by standard protocols, allowed the identification of more phytotoxic soils

  15. 天然气净化厂安全防火防爆系统的应用分析%Application and analysis of the safety and fire protection and explosion protection system of natural gas purification plant

    Institute of Scientific and Technical Information of China (English)



    本文就天然气净化厂安全防火防爆系统的应用进行分析,以促进天然气净化厂对安全工作的重视程度,提升安全管理水平。%In this paper, the application of natural gas purification plant safety fire and explosion protection system is analyzed, in order to promote the natural gas purification plant to the safety work of attention, improve the level of safety management.

  16. Purification of gibberellin sub 53 -oxidase from spinach

    Energy Technology Data Exchange (ETDEWEB)

    Wilson, T.M.; Zeevaart, J.A.D. (Michigan State Univ., East Lansing (USA))


    Spinach is a long-day rosette plants, in which stem growth is mediated by gibberellins. It has been shown that two enzymatic steps, GA{sub 53}-oxidase and GA{sub 19}-oxidase, are controlled by light. To develop an understanding into this light regulation, purification of GA{sub 53}-oxidase has been undertaken. The original assay relied on the HPLC separation of the product and substrate, but was considered too slow for the development of a purification scheme. A TLC system was developed which in conjunction with improvements to the assay conditions was sensitive and gave rapid results. The partial purification of the GA{sub 53}-oxidase is achieved by a high speed centrifugation, 40-55% ammonium sulfate precipitation, an hydroxyapatite column, Sephadex G-100 column and an anion exchange FPLC column, Mono Q HR10/10, yielding 1000-fold purification and 15% recovery. Monoclonal antibodies to the protein will be raised and used to further characterize the enzyme.

  17. Ecological Purification Efficiency of Several Aquatic Plants on Tail Water of Sewage Treatment Plant%不同水生植物对污水处理厂尾水的生态净化效果分析

    Institute of Scientific and Technical Information of China (English)



    通过构建小型水生生态系统,研究了旱伞草、美人蕉、伊乐藻、金鱼藻4种水生植物对太湖流域污水处理厂尾水中氮、磷等指标去除效能的差异. 结果表明,4种水生植物对污水中的氮、磷等均有明显的去除效果,其中,挺水植物旱伞草和沉水植物金鱼藻的综合净化效能较强,综合净化能力从强到弱依次为金鱼藻、旱伞草、伊乐藻、美人蕉.%Based on building the small aquatic ecosystem, the removal efficiency of nitrogen and phosphorus in sewage treatment plant tail water of Taihu Lake Basin by Cyperus alternifolius, Canna indica, Elodea Canadensis Michx.and Ceratophyllum demersum L.was studied.The results showed that these four plants had good removal efficiency for nitrogen and phosphorus in wastewater.The comprehensive purification ability of emerged plants Cyperus alternifolius and submerged plants Ceratophyllum demersum L.was higher than that of the others.The order of comprehensive purification ability from strong to weak was Ceratophyllum demersum L., Cyperus alternifolius, Elodea Canadensis Michx., Canna indica.

  18. Catalyst useful at highter temperatures, especially for purification of exhaust gases from motor vehicles and industrial plants

    Energy Technology Data Exchange (ETDEWEB)

    Koberstein, E.; Lakatos, E.


    A description is given of a catalyst for the purification of exhaust gases from industrial processes and motor vehicles by combustion of oxidizable impurities to carbon dioxide and water and by removal of nitrogen oxide at elevated temperatures in contact with air. The catalyst consists essentially of a calcined mixture of eta and gamma aluminum oxide and heavy metal oxide compounds containing chromium and nickel or copper with nickel. The metal oxide compounds are 55 to 90 weight percent, and the molar proportion of the chromium to the other heavy metal is between about 1:0.5 and 1:2.5. The metal oxide compounds are formed by precipitating chromium from a solution of ammonium bichromate with a soluble salt of the other metal or metals. Purification of exhaust gases from industrial processes and motor vehicles, as well as a method for making the catalyst, are discussed.


    Directory of Open Access Journals (Sweden)

    Deepinderjeet Singh Joshan


    Full Text Available The present study is concerned with both in-vitro assessment of antioxidant activity and anti-hemolytic effects of Ficus bengalensis, Calendula officinalis and Juglans regia. Total flavonoids and phenolics also were determined by using aluminum nitrate and Folin–Ciocalteu colorimetric methods respectively. The antioxidant capacity of sample was assessed through reducing power assay, DPPH-scavenging effect,metal chelation assay and superoxide scavenging assay. The extract of Calendula officinalis was found to be more efficient in as antioxidant and anti-hemolytic agents using the in vitro assays as compared to Ficus bengalensis and Juglans regia.

  20. Expression and Purification of PI3 Kinase {alpha} and Development of an ATP Depletion and an AlphaScreen PI3 Kinase Activity Assay

    DEFF Research Database (Denmark)

    Boldyreff, Brigitte; Rasmussen, Tine L; Jensen, Hans H


    -terminal His-tag and the p85alpha regulatory domain in Sf9 insect cells. The complex consisting of p110alpha and p85alpha was purified by nickel affinity chromatography. The authors established an adenosine triphosphate (ATP) depletion assay to measure the activity of p110alpha/p85alpha. The assay...

  1. A sensitive ferricyanide-mediated biochemical oxygen demand assay for analysis of wastewater treatment plant influents and treated effluents. (United States)

    Jordan, Mark A; Welsh, David T; John, Richard; Catterall, Kylie; Teasdale, Peter R


    Representative and fast monitoring of wastewater influent and effluent biochemical oxygen demand (BOD) is an elusive goal for the wastewater industry and regulatory bodies alike. The present study describes a suitable assay, which incorporates activated sludge as the biocatalyst and ferricyanide as the terminal electron acceptor for respiration. A number of different sludges and sludge treatments were investigated, primarily to improve the sensitivity of the assay. A limit of detection (LOD) (2.1 mg BOD₅ L⁻¹) very similar to that of the standard 5-day BOD₅ method was achieved in 4 h using raw influent sludge that had been cultured overnight as the biocatalyst. Reducing the microbial concentration was the most effective means to improve sensitivity and reduce the contribution of the sludge's endogenous respiration to total ferricyanide-mediated (FM) respiration. A strong and highly significant relationship was found (n = 33; R = 0.96; p BOD₅ and FM-BOD equivalent values for a diverse range of samples including wastewater treatment plant (WWTP) influent and treated effluent, as well as several grey water samples. The activated sludge FM-BOD assay presented here is an exceptional surrogate method to the standard BOD₅ assay, providing representative, same-day BOD analysis of WWTP samples with a comparable detection limit, a 4-fold greater analytical range and much faster analysis time. The industry appeal of such an assay is tremendous given that ~90% of all BOD₅ analysis is dedicated to measurement of WWTP samples, for which this assay is specifically designed.

  2. Removal of radioactive iodine and cesium in water purification processes after an explosion at a nuclear power plant due to the Great East Japan Earthquake. (United States)

    Kosaka, Koji; Asami, Mari; Kobashigawa, Naoya; Ohkubo, Keiko; Terada, Hiroshi; Kishida, Naohiro; Akiba, Michihiro


    The presence of radionuclides at five water purification plants was investigated after an explosion at a nuclear power plant hit by the Great East Japan Earthquake on 11 March 2011. Radioactive iodine (¹³¹I) and cesium (¹³⁴Cs and ¹³⁷Cs) were detected in raw water in Fukushima and neighboring prefectures. ¹³¹I was not removed by coagulation-flocculation-sedimentation. ¹³¹I was removed by granular activated carbon (GAC) and powdered activated carbon (PAC) at a level of about 30%-40%, although ¹³¹I was not removed in some cases. This was also confirmed by laboratory-scale experiments using PAC. The removal percentages of ¹³¹I in river and pond waters by 25 mg dry/L of PAC increased from 36% to 59% and from 41% to 48%, respectively, with chlorine dosing before PAC. ¹³⁴Cs and ¹³⁷Cs were effectively removed by coagulation at both a water purification plant and in laboratory-scale experiments when turbidity was relatively high. In contrast, ¹³⁴Cs and ¹³⁷Cs in pond water with low turbidity were not removed by coagulation. This was because ¹³⁴Cs and ¹³⁷Cs in river water were present mainly in particulate form, while in pond water they were present mainly as cesium ions (¹³⁴Cs+ and ¹³⁷Cs+). However, the removal of ¹³⁴Cs and ¹³⁷Cs in pond water by coagulation increased markedly when ¹³⁴Cs and ¹³⁷Cs were mixed with sediment 24 h before coagulation.

  3. 水生植物对铜污染废水的净化能力%Purification Capacity of Aquatic Plants for Treatment of Wastewater with Copper Contaminated

    Institute of Scientific and Technical Information of China (English)

    沈瑾; 王三反; 孙连鹏


    Canna (canna indiaca), arrowhead (saguttarua saguttefolia), calamus (acorns calamus linn), cattail (typhalatifolia), wild rice stem (zizania aquatica) were selected on the basis of their capacity of copper accumulation, to purify wastewater containing copper. The results show that more than 90 % of copper is removed in water; most of absorption of copper stores in plants' root, only a few is transported to stem and leaf; compared to copper purification capacity of cattail, copper purification capacity of arrowhead and wild rice stem are better. Arrowhead and wild rice stem can be integrated to purify wastewater containing copper.%该文选用了美人蕉、慈姑、菖蒲、香蒲和茭白五种对铜有一定富集能力的植物进行铜污染废水的净化效果研究,结果表明90%以上的铜得以从水中去除.植物吸收的铜大都贮存在根系中,只有极少一部分被传输转运到地上部分的茎、叶中.净化效果香蒲较差,以茭白和慈姑为好,可以将这2种水生植物综合用于植物修复.

  4. An effective high-speed countercurrent chromatographic method for preparative isolation and purification of mollugin directly from the ethanol extract of the Chinese medicinal plant Rubia cordifolia. (United States)

    Lu, Yanbin; Liu, Rui; Sun, Cuirong; Pan, Yuanjiang


    The medicinal plant Rubia cordifolia has been used widely in traditional Chinese medicine (TCM) for its antibacterial, antioxidant and anti-inflammatory activities. In this study, a preparative high-speed countercurrent chromatography (HSCCC) method for isolation and purification of the bioactive component mollugin directly from the ethanol extract of R. cordifolia was successfully established by using light petroleum (bp 60-90 degrees C)/ethanol/diethyl ether/water as the two-phase solvent system. The upper phase of light petroleum/ethanol/diethyl ether/water (5:4:3:1 v/v) was used as the stationary phase of HSCCC. Under the optimum conditions, 46 mg of mollugin at 98.5% purity, as determined by HPLC, could be yielded from 500 mg of the crude extract in a single HSCCC separation. The peak fraction of HSCCC was identified by 1H NMR and 13C NMR.

  5. A modified HET-CAM assay approach to the assessment of anti-irritant properties of plant extracts. (United States)

    Wilson, T D; Steck, W F


    Hen's egg--chorioallantoic membranes were used to screen for and assess anti-irritant properties among aqueous extracts of plants (HET-CAM tests), in connection with searches for plant-derived substances with topical anti-irritant action. The main question to be answered was whether CAM-assay screening of plant extracts could provide a useful route to identifying promising anti-irritant extracts for follow-up clinical testing. To be useful, the method would have to flag materials with strong anti-irritant properties, and would have to avoid registering false negatives. The tests conducted provided positive indications. We measured the delays in onset of three manifestations of membrane irritation-vascular hemorrhaging, membrane lysis and membrane coagulation-observed with test substances relative to positive controls. Aqueous 15% lactic acid, a commonly used irritant in direct tests on human skin, was employed as the test irritant in this study. The ratio [irritation onset times after test substance pre-treatment]:[onset times without test substance pretreatment] was used to measure the anti-irritant power of test substances. A scoring notation was devised for this which treats the delay parameters as independent effects. Most tested plant extracts showed no significant irritant or anti-irritant effects. Among the apparently anti-irritant plant extracts (approx. 10% of all those tested), most showed their greatest effect against hemorrhaging. Lesser but still readily measurable effects against membrane lysis and coagulation were also observed in nearly all the apparently anti-irritant extracts. Two of the tested extracts proved to be membrane irritants. Some key CAM assay results were compared with results obtained in direct tests on human skin using the same test irritant (15% lactic acid). In these comparative tests on skin, an essentially similar pattern of efficacy was obtained, with the plant extract deemed best in the CAM screenings, outperforming the

  6. Development of a plant viral-vector-based gene expression assay for the screening of yeast cytochrome p450 monooxygenases. (United States)

    Hanley, Kathleen; Nguyen, Long V; Khan, Faizah; Pogue, Gregory P; Vojdani, Fakhrieh; Panda, Sanjay; Pinot, Franck; Oriedo, Vincent B; Rasochova, Lada; Subramanian, Mani; Miller, Barbara; White, Earl L


    Development of a gene discovery tool for heterologously expressed cytochrome P450 monooxygenases has been inherently difficult. The activity assays are labor-intensive and not amenable to parallel screening. Additionally, biochemical confirmation requires coexpression of a homologous P450 reductase or complementary heterologous activity. Plant virus gene expression systems have been utilized for a diverse group of organisms. In this study we describe a method using an RNA vector expression system to phenotypically screen for cytochrome P450-dependent fatty acid omega-hydroxylase activity. Yarrowia lipolytica CYP52 gene family members involved in n-alkane assimilation were amplified from genomic DNA, cloned into a plant virus gene expression vector, and used as a model system for determining heterologous expression. Plants infected with virus vectors expressing the yeast CYP52 genes (YlALK1-YlALK7) showed a distinct necrotic lesion phenotype on inoculated plant leaves. No phenotype was detected on negative control constructs. YlALK3-, YlALK5-, and YlALK7-inoculated plants all catalyzed the terminal hydroxylation of lauric acid as confirmed using thin-layer and gas chromatography/mass spectrometry methods. The plant-based cytochrome P450 phenotypic screen was tested on an n-alkane-induced Yarrowia lipolytica plant virus expression library. A subset of 1,025 random library clones, including YlALK1-YlALK7 constructs, were tested on plants. All YlALK gene constructs scored positive in the randomized screen. Following nucleotide sequencing of the clones that scored positive using a phenotypic screen, approximately 5% were deemed appropriate for further biochemical analysis. This report illustrates the utility of a plant-based system for expression of heterologous cytochrome P450 monooxygenases and for the assignment of gene function.

  7. Arabidopsis seedling flood-inoculation technique: a rapid and reliable assay for studying plant-bacterial interactions

    Directory of Open Access Journals (Sweden)

    Uppalapati Srinivasa R


    Full Text Available Abstract Background The Arabidopsis thaliana-Pseudomonas syringae model pathosystem is one of the most widely used systems to understand the mechanisms of microbial pathogenesis and plant innate immunity. Several inoculation methods have been used to study plant-pathogen interactions in this model system. However, none of the methods reported to date are similar to those occurring in nature and amicable to large-scale mutant screens. Results In this study, we developed a rapid and reliable seedling flood-inoculation method based on young Arabidopsis seedlings grown on MS medium. This method has several advantages over conventional soil-grown plant inoculation assays, including a shorter growth and incubation period, ease of inoculation and handling, uniform infection and disease development, requires less growth chamber space and is suitable for high-throughput screens. In this study we demonstrated the efficacy of the Arabidopsis seedling assay to study 1 the virulence factors of P. syringae pv. tomato DC3000, including type III protein secretion system (TTSS and phytotoxin coronatine (COR; 2 the effector-triggered immunity; and 3 Arabidopsis mutants affected in salicylic acid (SA- and pathogen-associated molecular pattern (PAMPs-mediated pathways. Furthermore, we applied this technique to study nonhost resistance (NHR responses in Arabidopsis using nonhost pathogens, such as P. syringae pv. tabaci, pv. glycinea and pv. tomato T1, and confirmed the functional role of FLAGELLIN-SENSING 2 (FLS2 in NHR. Conclusions The Arabidopsis seedling flood-inoculation assay provides a rapid, efficient and economical method for studying Arabidopsis-Pseudomonas interactions with minimal growth chamber space and time. This assay could also provide an excellent system for investigating the virulence mechanisms of P. syringae. Using this method, we demonstrated that FLS2 plays a critical role in conferring NHR against nonhost pathovars of P. syringae, but not to

  8. A trypsin-like proteinase in the midgut of Ectomyelois ceratoniae Zeller (Lepidoptera: Pyralidae): purification, characterization, and host plant inhibitors. (United States)

    Ranjbar, Mina; Zibaee, Arash; Sendi, Jalal Jalali


    A trypsin-like proteinase was purified and characterized in the midgut of Ectomyelois ceratoniae. A purification process that used Sepharyl G-100 and DEAE-cellulose fast flow chromatographies revealed a proteinase with specific activity of 66.7 μmol/min/mg protein, recovery of 27.04 and purification fold of 23.35. Molecular weight of the purified protein was found to be 35.8 kDa. Optimal pH and temperature were obtained 9 and 20°C for the purified trypsin proteinase, respectively. The purified enzyme was significantly inhibited by PMSF, TLCK, and SBTI as specific inhibitors of trypsins in which TLCK showed the highest inhibitory effect. Trypsin proteinase inhibitors were extracted from four varieties of pomegranate including Brait, Torsh-Sabz, May-Khosh, and Shirin by ion exchange chromatography. It was found that fractions 17-20 of Brait; fractions 18 and 21-26 of Torsh-Sabz; fractions 1-7, 11-17, and 19-21 of May-Khosh and fraction 8 for Shirin showed presence of trypsin inhibitor in these host. Comparison of their inhibitory effects on the purified trypsin proteinase of E. ceratoniae demonstrated that fractions from May-khosh variety had the highest effect on the enzyme among other extracted fractions. Characterization of serine proteinases of insects mainly trypsins is one of the promising methods to decrease population and damages via extracting their inhibitors and providing resistant varieties.

  9. Molecular Characterization of the Bacterial Communities in the Different Compartments of a Full-Scale Reverse-Osmosis Water Purification Plant (United States)

    Bereschenko, L. A.; Heilig, G. H. J.; Nederlof, M. M.; van Loosdrecht, M. C. M.; Stams, A. J. M.; Euverink, G. J. W.


    The origin, structure, and composition of biofilms in various compartments of an industrial full-scale reverse-osmosis (RO) membrane water purification plant were analyzed by molecular biological methods. Samples were taken when the RO installation suffered from a substantial pressure drop and decreased production. The bacterial community of the RO membrane biofilm was clearly different from the bacterial community present at other locations in the RO plant, indicating the development of a specialized bacterial community on the RO membranes. The typical freshwater phylotypes in the RO membrane biofilm (i.e., Proteobacteria, Cytophaga-Flexibacter-Bacteroides group, and Firmicutes) were also present in the water sample fed to the plant, suggesting a feed water origin. However, the relative abundances of the different species in the mature biofilm were different from those in the feed water, indicating that the biofilm was actively formed on the RO membrane sheets and was not the result of a concentration of bacteria present in the feed water. The majority of the microorganisms (59% of the total number of clones) in the biofilm were related to the class Proteobacteria, with a dominance of Sphingomonas spp. (27% of all clones). Members of the genus Sphingomonas seem to be responsible for the biofouling of the membranes in the RO installation. PMID:18621875

  10. Development and qualification of a high sensitivity, high throughput Q-PCR assay for quantitation of residual host cell DNA in purification process intermediate and drug substance samples. (United States)

    Zhang, Wei; Wu, Meng; Menesale, Emily; Lu, Tongjun; Magliola, Aeona; Bergelson, Svetlana


    Methods of high sensitivity, accuracy and throughput are needed for quantitation of low level residual host cell DNA in purification process intermediates and drug substances of therapeutic proteins. In this study, we designed primer/probe sets targeting repetitive Alu repeats or Alu-equivalent sequences in the human, Chinese hamster and murine genomes. When used in quantitative polymerase chain reactions (Q-PCRs), these primer/probe sets showed high species specificity and gave significantly higher sensitivity compared to those targeting the low copy number GAPDH gene. This allowed for detection of residual host cell DNA of much lower concentrations and, for some samples, eliminated the need for DNA extraction. By combining the high sensitivity Alu Q-PCR with high throughput automated DNA extraction using an automated MagMAX magnetic particle processor, we successfully developed and qualified a highly accurate, specific, sensitive and efficient method for the quantitation of residual host cell DNA in process intermediates and drug substances of multiple therapeutic proteins purified from cells of multiple species. Compared to the previous method using manual DNA extraction and primer/probe sets targeting the GAPDH gene, this new method increased our DNA extraction throughput by over sevenfold, and lowered the lower limit of quantitation by up to eightfold.

  11. The species and distribution of air purification plants in Shijiazhuang urban area%石家庄市区空气净化植物的种类及分布研究

    Institute of Scientific and Technical Information of China (English)

    石金秋; 万五星; 宋雅丽


    By the investigation of the wild and cultivated plants in Shijiazhuang, the species, distribution and functions of the air purification plants are analyzed. Based on the results a number of recommendations about development, utilization and protection of air purification plants in Shijiazhuang urban area are provided.%通过对石家庄市区野生和栽培植物的调查,对有空气净化作用的植物种类、分布及功能进行分析,提出了石家庄市空气净化植物的开发利用和保护发展的若干建议.

  12. In vitro Antimicrobial Assay of Actinomycetes in Rice AgainstXanthomonas oryzae pv. oryzicola and as Potential Plant Growth Promoter

    Directory of Open Access Journals (Sweden)

    Erneeza Mohd Hata


    Full Text Available ABSTRACT The aim of this work was to invitro assay the antimicrobial activity of actinomycetes in rice against Xanthomonas oryzae pv. oryzicola and as potential plant growth promoter. A total of 92 actinomycete strains were isolated from different rice plant components and field locations. Of these, only 21.74% showed antagonistic activity against the Xoc pathogen. Molecular identification via 16s rRNA amplification revealed that 60% of the active antagonistic strains belonged to the genus Streptomyces. Isolates that demonstrated the highest antagonistic activity were also able to produce hydrolytic enzymes and plant growth-promoting hormones. Combination of preliminary screening based on in vitro antagonistic, hydrolytic enzyme and plant growth hormone activity facilitated the best selection of actinomycete candidates as evidenced by strains classification using cluster analysis (Ward's Method. Results from the preliminary screening showed that actinomycetes, especially Streptomycetes, could offer a promising source for both biocontrol and plant growth-promotion agents against BLS disease in rice.

  13. Isolation and Purification of Oridonin from the Whole Plant of Isodon rubescens by High-Speed Counter-Current Chromatography

    Directory of Open Access Journals (Sweden)

    ChunYue Yu


    Full Text Available Semi-preparative high-speed counter-current chromatography (HSCCC was successfully used for isolation and purification of oridonin from Isodon rubescens by using a two-phase-solvent system composed of n-hexane-ethyl acetate-methanol-water (2.8:5:2.8:5, v/v/v/v. The targeted compound isolated, collected and purified by HSCCC was analyzed by high performance liquid chromatography (HPLC. A total of 40.6 mg of oridonin with the purity of 73.5% was obtained in less than 100 min from 100 mg of crude Isodon rubescens extract. The chemical structure of the compound was identified by IR, 1H-NMR and 13C-NMR.

  14. Hamiltonian purification

    Energy Technology Data Exchange (ETDEWEB)

    Orsucci, Davide [Scuola Normale Superiore, I-56126 Pisa (Italy); Burgarth, Daniel [Department of Mathematics, Aberystwyth University, Aberystwyth SY23 3BZ (United Kingdom); Facchi, Paolo; Pascazio, Saverio [Dipartimento di Fisica and MECENAS, Università di Bari, I-70126 Bari (Italy); INFN, Sezione di Bari, I-70126 Bari (Italy); Nakazato, Hiromichi; Yuasa, Kazuya [Department of Physics, Waseda University, Tokyo 169-8555 (Japan); Giovannetti, Vittorio [NEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR, I-56126 Pisa (Italy)


    The problem of Hamiltonian purification introduced by Burgarth et al. [Nat. Commun. 5, 5173 (2014)] is formalized and discussed. Specifically, given a set of non-commuting Hamiltonians (h{sub 1}, …, h{sub m}) operating on a d-dimensional quantum system ℋ{sub d}, the problem consists in identifying a set of commuting Hamiltonians (H{sub 1}, …, H{sub m}) operating on a larger d{sub E}-dimensional system ℋ{sub d{sub E}} which embeds ℋ{sub d} as a proper subspace, such that h{sub j} = PH{sub j}P with P being the projection which allows one to recover ℋ{sub d} from ℋ{sub d{sub E}}. The notions of spanning-set purification and generator purification of an algebra are also introduced and optimal solutions for u(d) are provided.

  15. Enzyme-linked immunosorbent assay for the detection and identification of plant pathogenic bacteria (in particular for Erwinia amylovora and Clavibacter michiganensis subsp. sepedonicus). (United States)

    Kokoskova, Blanka; Janse, Jaap D


    Enzyme-linked immunosorbent assay (ELISA) is the most commonly used serological diagnostic technique. A number of different ELISA formats can be used for the detection of bacterial plant pathogens and in particular Erwinia amylovora and Clavibacter michiganensis subsp. sepedonicus.

  16. Ribosome-inactivating proteins in edible plants and purification and characterization of a new ribosome-inactivating protein from Cucurbita moschata. (United States)

    Barbieri, Luigi; Polito, Letizia; Bolognesi, Andrea; Ciani, Marialibera; Pelosi, Emanuele; Farini, Valentina; Jha, Ajay K; Sharma, Neelam; Vivanco, Jorge M; Chambery, Angela; Parente, Augusto; Stirpe, Fiorenzo


    The basic protein fraction of tissue extracts from 40 edible plants inhibited cell-free protein synthesis and released adenine from herring sperm DNA, thus having adenine glycosylase activity. This suggested the presence of ribosome-inactivating proteins (RIPs) in the plant extracts. This indication was further strengthened by the presence of the two activities after a partial chromatographic purification of three extracts, including that from Lycopersicon esculentum (tomato), which had very low activity. From the extract of Cucurbita moschata (pumpkin), the most active one, a glycoprotein of 30,665 Da was purified which had the properties of a RIP, in that (i) it inhibited protein synthesis by a rabbit reticulocyte lysate with IC50 (concentration giving 50% inhibition) 0.035 nM (1.08 ng ml(-1)) and by HeLa, HT29 and JM cells with IC50 in the 100 nM range, (ii) deadenylated hsDNA and other polynucleotidic substrates, and (iii) depurinated yeast rRNA at a concentration of 0.1 ng ml(-1), all values being comparable to those of other RIPs. The C. moschata RIP gave a weak cross-reaction only with an antiserum against dianthin 32, but not with antisera against other RIPs, and had superoxide dismutase, antifungal and antibacterial activities.

  17. Loop-mediated Isothermal Amplification Assay to Rapidly Detect Wheat Streak Mosaic Virus in Quarantined Plants

    Directory of Open Access Journals (Sweden)

    Siwon Lee


    Full Text Available We developed a loop-mediated isothermal amplification (LAMP method to rapidly diagnose Wheat streak mosaic virus (WSMV during quarantine inspections of imported wheat, corn, oats, and millet. The LAMP method was developed as a plant quarantine inspection method for the first time, and its simplicity, quickness, specificity and sensitivity were verified compared to current reverse transcription-polymerase chain reaction (RT-PCR and nested PCR quarantine methods. We were able to quickly screen for WSMV at quarantine sites with many test samples; thus, this method is expected to contribute to plant quarantine inspections.

  18. A rapid and robust assay for detection of S-phase cell cycle progression in plant cells and tissues by using ethynyl deoxyuridine

    Directory of Open Access Journals (Sweden)

    Horváth Gábor V


    Full Text Available Abstract Background Progress in plant cell cycle research is highly dependent on reliable methods for detection of cells replicating DNA. Frequency of S-phase cells (cells in DNA synthesis phase is a basic parameter in studies on the control of cell division cycle and the developmental events of plant cells. Here we extend the microscopy and flow cytometry applications of the recently developed EdU (5-ethynyl-2'-deoxyuridine-based S-phase assay to various plant species and tissues. We demonstrate that the presented protocols insure the improved preservation of cell and tissue structure and allow significant reduction in assay duration. In comparison with the frequently used detection of bromodeoxyuridine (BrdU and tritiated-thymidine incorporation, this new methodology offers several advantages as we discuss here. Results Applications of EdU-based S-phase assay in microscopy and flow cytometry are presented by using cultured cells of alfalfa, Arabidopsis, grape, maize, rice and tobacco. We present the advantages of EdU assay as compared to BrdU-based replication assay and demonstrate that EdU assay -which does not require plant cell wall digestion or DNA denaturation steps, offers reduced assay duration and better preservation of cellular, nuclear and chromosomal morphologies. We have also shown that fast and efficient EdU assay can also be an efficient tool for dual parameter flow cytometry analysis and for quantitative assessment of replication in thick root samples of rice. Conclusions In plant cell cycle studies, EdU-based S-phase detection offers a superior alternative to the existing S-phase assays. EdU method is reliable, versatile, fast, simple and non-radioactive and it can be readily applied to many different plant systems.

  19. Standard assays do not predict the efficiency of commercial cellulase preparations towards plant materials

    NARCIS (Netherlands)

    Kabel, M.A.; Maarel, van der M.J.E.C.; Klip, G.; Voragen, A.G.J.; Schols, H.A.


    Commercial cellulase preparations are potentially effective for processing biomass feedstocks in order to obtain bioethanol. In plant cell walls, cellulose fibrils occur in close association with xylans (monocotyls) or xyloglucans (dicotyls). The enzymatic conversion of cellulose/xylans is a complex

  20. Standard Assays Do Not Predict the Efficiency of Commercial Cellulase Preparations Towards Plant Materials

    NARCIS (Netherlands)

    Kabel, Mirjam A.; Maarel, Marc J.E.C. van der; Klip, Gert; Voragen, Alphons G.J.; Schols, Henk A.


    Commercial cellulase preparations are potentially effective for processing biomass feedstocks in order to obtain bioethanol. In plant cell walls, cellulose fibrils occur in close association with xylans (monocotyls) or xyloglucans (dicotyls). The enzymatic conversion of cellulose/xylans is a complex

  1. Standard assays do not predict the efficiency of commercial cellulase preparations towards plant materials

    NARCIS (Netherlands)

    Kabel, M.A.; Maarel, van der M.J.E.C.; Klip, G.; Voragen, A.G.J.; Schols, H.A.


    Commercial cellulase preparations are potentially effective for processing biomass feedstocks in order to obtain bioethanol. In plant cell walls, cellulose fibrils occur in close association with xylans (monocotyls) or xyloglucans (dicotyls). The enzymatic conversion of cellulose/xylans is a complex

  2. Standard Assays Do Not Predict the Efficiency of Commercial Cellulase Preparations Towards Plant Materials

    NARCIS (Netherlands)

    Kabel, Mirjam A.; Maarel, Marc J.E.C. van der; Klip, Gert; Voragen, Alphons G.J.; Schols, Henk A.


    Commercial cellulase preparations are potentially effective for processing biomass feedstocks in order to obtain bioethanol. In plant cell walls, cellulose fibrils occur in close association with xylans (monocotyls) or xyloglucans (dicotyls). The enzymatic conversion of cellulose/xylans is a complex

  3. Studies on luciferin-luciferase ATP assay in plants (etiolated wheat germs, and bean leaves

    Directory of Open Access Journals (Sweden)

    A. Barbaro


    Full Text Available For ATP determination by the method of bioluminescence apparatus of home production was adapted from the equipment available in any isotope laboratory. The measurement error did not exceed 1.5 per cent. Methodical experiments concerned the choice of the extraction, fixation and storage methods of plant material for determination at the given moment of the amount of ATP in the tissues, unchanged by the analytical procedure. The highest ATP amounts were recovered by extraction with perchloric acid at high (25% concentrations of the tissue in the homogenate. The best way of fixation of the material for later analyses was found to be freezing of ready extracts. Lyophilization and freezing of the plant material caused a several-fold decrease of the ATP level in the tissues. These results suggest the necessity of working in conditions of low temperature during the entire analytical procedure and strict observation of time limitation.

  4. Drug Development and Conservation of Biodiversity in West and Central Africa: Performance of Neurochemical and Radio Receptor Assays of Plant Extracts Drug Discovery for the Central Nervous System (United States)


    not so many years ago that single drug substances resulting from rational design and synthesis would replace the old fashioned herbal plants which...1.2 g) eluted with hexane-EtOAc (8:2) was then passed through a silica gel column using CH2Cl2 to give a mixture that was further purified on...purified in the same way as frac- tion F3 to yield TS1 (40 mg) and TS3 (133 mg). An additional purification of TS1 and TS3 by gel per- meation through

  5. Development of a sensitive E-screen assay for quantitative analysis of estrogenic activity in municipal sewage plant effluents. (United States)

    Körner, W; Hanf, V; Schuller, W; Kempter, C; Metzger, J; Hagenmaier, H


    A simplified proliferation test with human estrogen receptor-positive MCF-7 breast cancer cells (E-screen assay) was optimized and validated for the sensitive quantitative determination of total estrogenic activity in effluent samples from municipal sewage plants. After solid phase extraction of 1 l sewage on either 0.2 g polystyrene copolymer (ENV+) or 1 g RP-C18 material and removal of the solvent, analysis of the extracts in the E-screen assay could be performed without any clean-up step. This was even possible with untreated sewage. Parallel extraction of four sewage samples on both different solid phase materials gave comparable quantitative results in the E-screen. A blank sample did not induce cell proliferation. As additive behaviour of the estrogenic response of single compounds was proven for two different mixtures each containing three xenoestrogens, total estrogenic activity in the sewage samples, expressed as 17 beta-estradiol equivalent concentration (EEQ), could be calculated comparing the EC50 values of the samples with those of the positive control 17 beta-estradiol. The detection limit of the E-screen method was 0.05 pmol EEQ/l (0.014 ng EEQ/l), the limit of quantification 0.25-0.5 pmol EEQ/l (0.07-0.14 ng EEQ/l). In total, extracts of nine effluent and one influent sample from five different municipal sewage plants in South Germany were analyzed in the E-screen. All samples strongly induced cell proliferation in a dose-dependent manner which was completely inhibited by coincubation with 5 nM of the estrogen receptor-antagonist ICI 182,780. The proliferative effect relative to the positive control 17 beta-estradiol (RPE) was between 30 and 101%. 17 beta-Estradiol equivalent concentrations were between 2.5 and 25 ng/l indicating a significant input of estrogenic substances via sewage treatment plants into rivers.

  6. Review on production run and technical transformation of Wanzhou Natural Gas Purification Plant%万州天然气净化厂生产运行及技术改造评价

    Institute of Scientific and Technical Information of China (English)

    廖铁; 李法璋; 龚毅然; 苏梦瑶; 夏平琼


    重庆天然气净化总厂万州分厂于2009年6月进气投产,装置平稳运行至今,达到了工程项目设计要求,但在生产运行过程中也出现了一些问题。为了进一步掌握该装置的运行情况,结合万州分厂近5年的生产运行情况,重点介绍了装置的生产运行特点及存在的问题,并提出对应的改进措施,可为其他天然气净化厂的设计及安全生产运行提供技术参考。%Since being put into operation in June 2009 ,the device of Wanzhou Natural Gas Purification Plant has got stable operation and reached design requirements . However , some problems occurred during the production .In combination with 5 year's operation situation of the purification plant ,the operation characteristics of the process were introduced ,some problems occurring during the production were analyzed ,and the corresponding improvement measures were proposed ,which could provide some references for the design and safe production of similar natural gas purification plants .

  7. Char characterization and DTF assays as tools to predict burnout of coal blends in power plants

    Energy Technology Data Exchange (ETDEWEB)

    C. Ulloa; A.G. Borrego; S. Helle; A.L. Gordon; X. Garcia [Universidad de Concepcion, Concepcion (Chile). Departamento de Ingenieria Quimica


    The aim of this study is to predict efficiency deviations in the combustion of coal blends in power plants. Combustion of blends, as compared to its single coals, shows that for some blends the behavior is non-additive in nature. Samples of coal feed and fly ashes from combustion of blends at two power plants, plus chars of the parent coals generated in a drop-tube furnace (DTF) at temperatures and heating rates similar to those found in the industrial boilers were used. Intrinsic kinetic parameters, burning profiles and petrographic characteristics of these chars correlated well with the burnout in power plants and DTF experiments. The blend combustion in a DTF reproduces both positive and negative burnout deviations from the expected weighted average. These burnout deviations have been previously attributed to parallel or parallel-series pathways of competition for oxygen. No deviations were found for blends of low rank coals of similar characteristics yielding chars close in morphology, optical texture and reactivity. Negative deviations were found for blends of coals differing moderately in rank and were interpreted as associated with long periods of competition. In this case, fly-ashes were enriched in material derived from the least reactive char, but also unburnt material attributed to the most reactive char was identified. Improved burnout compared to the weighted average was observed for blends of coals very different in rank, and interpreted as the result of a short interaction period, followed by a period where the less reactive char burns under conditions that are more favorable to its combustion. In this case, only unburned material from the least reactive char was identified in the fly-ashes. 20 refs., 9 figs., 5 tabs.

  8. Fungal disease detection in plants: Traditional assays, novel diagnostic techniques and biosensors. (United States)

    Ray, Monalisa; Ray, Asit; Dash, Swagatika; Mishra, Abtar; Achary, K Gopinath; Nayak, Sanghamitra; Singh, Shikha


    Fungal diseases in commercially important plants results in a significant reduction in both quality and yield, often leading to the loss of an entire plant. In order to minimize the losses, it is essential to detect and identify the pathogens at an early stage. Early detection and accurate identification of pathogens can control the spread of infection. The present article provides a comprehensive overview of conventional methods, current trends and advances in fungal pathogen detection with an emphasis on biosensors. Traditional techniques are the "gold standard" in fungal detection which relies on symptoms, culture-based, morphological observation and biochemical identifications. In recent times, with the advancement of biotechnology, molecular and immunological approaches have revolutionized fungal disease detection. But the drawback lies in the fact that these methods require specific and expensive equipments. Thus, there is an urgent need for rapid, reliable, sensitive, cost effective and easy to use diagnostic methods for fungal pathogen detection. Biosensors would become a promising and attractive alternative, but they still have to be subjected to some modifications, improvements and proper validation for on-field use. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Polonium purification

    Energy Technology Data Exchange (ETDEWEB)

    Baker, J.D.


    Three processes for the purification of {sup 210}Po from irradiated bismuth targets are described. Safety equipment includes shielded hotcells for the initial separation from other activation products, gloveboxes for handling the volatile and highly toxic materials, and provisions for ventilation. All chemical separations must be performed under vacuum or in inerted systems. Two of the processes require large amounts of electricity; the third requires vessels made from exotic materials.

  10. [Purification of eutrophic wastewater by Cyperus alternifolius, Coleus blumei and Jasminum sambac planted in a floating phytoremediation system]. (United States)

    Liu, Shizhe; Lin, Dongjiao; Tang, Shujun; Luo, Jian


    In a greenhouse study, Cyperus alternifolius, Coleus blumei and Jasminum sambac were cultured in a floating phytoremediation system with plantation cups inserted into a polyfoam plate that floated in the upper part of a tank filled with 100 L domestic wastewater. The contents of chemical oxygen demand (CODCr), total P (T-P), total N (T-N), soluble P(S-P), ammonia-nitrogen (NH4+ -N) and nitrate-nitrogen (NO3- -N) in the domestic wastewater were tested during the growth of these three plants. The results showed that Cyperus alternifolius and Coleus blumei could grow well in the floating phytoremediation system, their dry weight being 285.8% and 371.4% of the initial weight of planting, respectively, but Jasminum sambac could not grow well, being 125.0% of the initial weight of planting. The removal rate of TN by these 3 plants was 68.0%, 62.0% and 45.0%, and that of NO3- -N, CODCr and TP was 98.0%, 80.0% and 92.0%, 78.0%, 66.0% and 55.0%, and 90.6%, 90.5% and 88.0% respectively. Cyperus alternifolius and Coleus blumei had good effects on the removal of pollutants in the floating phytoremediation system.

  11. Chromosome analysis of nuclear power plant workers using fluorescence in situ hybridization and Giemsa assay (United States)

    Hristova, Rositsa; Hadjidekova, Valeria; Grigorova, Mira; Nikolova, Teodora; Bulanova, Minka; Popova, Ljubomira; Staynova, Albena; Benova, Donka


    The aim of this study was to evaluate the genotoxic effects of ionizing radiation in vivo in exposed Bulgarian nuclear power plant workers by using classical cytogenetic and molecular cytogenetic analyses of peripheral lymphocytes. Chromosome analysis using fluorescence in situ hybrydization (FISH) and Giemsa techniques was undertaken on 63 workers and 45 administrative staff controls from the Bulgarian Nuclear Power Plant. Using the Giemsa method, the frequencies of cells studied with chromosome aberrations, dicentrics plus rings and chromosome fragments in the radiation workers were significantly higher compared with the control group (P = 0.044, P = 0.014, and P = 0.033, respectively). A significant association between frequencies of dicentrics plus rings and accumulated doses was registered (P < 0.01). In the present study, a FISH cocktail of whole chromosome paints for chromosomes 1, 4 and 11 was used. A significant association between frequency of translocations and accumulated doses was also observed (P < 0.001). Within the control group, a correlation was found between age and the spontaneous frequency of translocations. No correlation was found between smoking status and frequency of translocations. When compared with the control group, workers with accumulated doses up to 100 mSv showed no increase in genome translocation frequency, whereas workers with accumulated doses from 101 to 200 mSv showed a statistically significant doubling of genome translocation frequency (P = 0.009). Thus, in cases of chronic exposure and for purposes of retrospective dosimetry, the genome frequency of translocations is a more useful marker for evaluation of genotoxic effects than dicentric frequency. PMID:23536543

  12. Chromosome analysis of nuclear power plant workers using fluorescence in situ hybridization and Giemsa assay. (United States)

    Hristova, Rositsa; Hadjidekova, Valeria; Grigorova, Mira; Nikolova, Teodora; Bulanova, Minka; Popova, Ljubomira; Staynova, Albena; Benova, Donka


    The aim of this study was to evaluate the genotoxic effects of ionizing radiation in vivo in exposed Bulgarian nuclear power plant workers by using classical cytogenetic and molecular cytogenetic analyses of peripheral lymphocytes. Chromosome analysis using fluorescence in situ hybrydization (FISH) and Giemsa techniques was undertaken on 63 workers and 45 administrative staff controls from the Bulgarian Nuclear Power Plant. Using the Giemsa method, the frequencies of cells studied with chromosome aberrations, dicentrics plus rings and chromosome fragments in the radiation workers were significantly higher compared with the control group (P = 0.044, P = 0.014, and P = 0.033, respectively). A significant association between frequencies of dicentrics plus rings and accumulated doses was registered (P < 0.01). In the present study, a FISH cocktail of whole chromosome paints for chromosomes 1, 4 and 11 was used. A significant association between frequency of translocations and accumulated doses was also observed (P < 0.001). Within the control group, a correlation was found between age and the spontaneous frequency of translocations. No correlation was found between smoking status and frequency of translocations. When compared with the control group, workers with accumulated doses up to 100 mSv showed no increase in genome translocation frequency, whereas workers with accumulated doses from 101 to 200 mSv showed a statistically significant doubling of genome translocation frequency (P = 0.009). Thus, in cases of chronic exposure and for purposes of retrospective dosimetry, the genome frequency of translocations is a more useful marker for evaluation of genotoxic effects than dicentric frequency.

  13. Thorium, uranium and rare earth elements content in lanthanide concentrate (LC) and water leach purification (WLP) residue of Lynas advanced materials plant (LAMP) (United States)

    AL-Areqi, Wadeeah M.; Majid, Amran Ab.; Sarmani, Sukiman


    Lynas Advanced Materials Plant (LAMP) has been licensed to produce the rare earths elements since early 2013 in Malaysia. LAMP processes lanthanide concentrate (LC) to extract rare earth elements and subsequently produce large volumes of water leach purification (WLP) residue containing naturally occurring radioactive material (NORM). This residue has been rising up the environmental issue because it was suspected to accumulate thorium with significant activity concentration and has been classified as radioactive residue. The aim of this study is to determine Th-232, U-238 and rare earth elements in lanthanide concentrate (LC) and water leach purification (WLP) residue collected from LAMP and to evaluate the potential radiological impacts of the WLP residue on the environment. Instrumental Neutron Activation Analysis and γ-spectrometry were used for determination of Th, U and rare earth elements concentrations. The results of this study found that the concentration of Th in LC was 1289.7 ± 129 ppm (5274.9 ± 527.6Bq/kg) whereas the Th and U concentrations in WLP were determined to be 1952.9±17.6 ppm (7987.4 ± 71.9 Bq/kg) and 17.2 ± 2.4 ppm respectively. The concentrations of Th and U in LC and WLP samples determined by γ- spectrometry were 1156 ppm (4728 ± 22 Bq/kg) & 18.8 ppm and 1763.2 ppm (7211.4 Bq/kg) &29.97 ppm respectively. This study showed that thorium concentrations were higher in WLP compare to LC. This study also indicate that WLP residue has high radioactivity of 232Th compared to Malaysian soil natural background (63 - 110 Bq/kg) and come under preview of Act 304 and regulations. In LC, the Ce and Nd concentrations determined by INAA were 13.2 ± 0.6% and 4.7 ± 0.1% respectively whereas the concentrations of La, Ce, Nd and Sm in WLP were 0.36 ± 0.04%, 1.6%, 0.22% and 0.06% respectively. This result showed that some amount of rare earth had not been extracted and remained in the WLP and may be considered to be reextracted.

  14. Thorium, uranium and rare earth elements content in lanthanide concentrate (LC) and water leach purification (WLP) residue of Lynas advanced materials plant (LAMP)

    Energy Technology Data Exchange (ETDEWEB)

    AL-Areqi, Wadeeah M., E-mail:; Majid, Amran Ab., E-mail:; Sarmani, Sukiman, E-mail: [Nuclear Science Programme, School of Chemical Sciences and Food Technology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi (Malaysia)


    Lynas Advanced Materials Plant (LAMP) has been licensed to produce the rare earths elements since early 2013 in Malaysia. LAMP processes lanthanide concentrate (LC) to extract rare earth elements and subsequently produce large volumes of water leach purification (WLP) residue containing naturally occurring radioactive material (NORM). This residue has been rising up the environmental issue because it was suspected to accumulate thorium with significant activity concentration and has been classified as radioactive residue. The aim of this study is to determine Th-232, U-238 and rare earth elements in lanthanide concentrate (LC) and water leach purification (WLP) residue collected from LAMP and to evaluate the potential radiological impacts of the WLP residue on the environment. Instrumental Neutron Activation Analysis and γ-spectrometry were used for determination of Th, U and rare earth elements concentrations. The results of this study found that the concentration of Th in LC was 1289.7 ± 129 ppm (5274.9 ± 527.6Bq/kg) whereas the Th and U concentrations in WLP were determined to be 1952.9±17.6 ppm (7987.4 ± 71.9 Bq/kg) and 17.2 ± 2.4 ppm respectively. The concentrations of Th and U in LC and WLP samples determined by γ- spectrometry were 1156 ppm (4728 ± 22 Bq/kg) and 18.8 ppm and 1763.2 ppm (7211.4 Bq/kg) and 29.97 ppm respectively. This study showed that thorium concentrations were higher in WLP compare to LC. This study also indicate that WLP residue has high radioactivity of {sup 232}Th compared to Malaysian soil natural background (63 - 110 Bq/kg) and come under preview of Act 304 and regulations. In LC, the Ce and Nd concentrations determined by INAA were 13.2 ± 0.6% and 4.7 ± 0.1% respectively whereas the concentrations of La, Ce, Nd and Sm in WLP were 0.36 ± 0.04%, 1.6%, 0.22% and 0.06% respectively. This result showed that some amount of rare earth had not been extracted and remained in the WLP and may be considered to be reextracted.

  15. Air/Water Purification (United States)


    After 18 years of research into air/water pollution at Stennis Space Center, Dr. B. C. Wolverton formed his own company, Wolverton Environmental Services, Inc., to provide technology and consultation in air and water treatment. Common houseplants are used to absorb potentially harmful materials from bathrooms and kitchens. The plants are fertilized, air is purified, and wastewater is converted to clean water. More than 100 U.S. communities have adopted Wolverton's earlier water hyacinth and artificial marsh applications. Catfish farmers are currently evaluating the artificial marsh technology as a purification system.

  16. Purification, cDNA cloning, and characterization of LysM-containing plant chitinase from horsetail (Equisetum arvense). (United States)

    Inamine, Saki; Onaga, Shoko; Ohnuma, Takayuki; Fukamizo, Tamo; Taira, Toki


    Chitinase-A (EaChiA), molecular mass 36 kDa, was purified from the vegetative stems of a horsetail (Equisetum arvense) using a series of column chromatography. The N-terminal amino acid sequence of EaChiA was similar to the lysin motif (LysM). A cDNA encoding EaChiA was cloned by rapid amplification of cDNA ends and polymerase chain reaction. It consisted of 1320 nucleotides and encoded an open reading frame of 361 amino acid residues. The deduced amino acid sequence indicated that EaChiA is composed of a N-terminal LysM domain and a C-terminal plant class IIIb chitinase catalytic domain, belonging to the glycoside hydrolase family 18, linked by proline-rich regions. EaChiA has strong chitin-binding activity, however, no antifungal activity. This is the first report of a chitinase from Equisetopsida, a class of fern plants, and the second report of a LysM-containing chitinase from a plant.

  17. Evaluation of acute toxicity and teratogenic effects of plant growth regulators by Daphnia magna embryo assay. (United States)

    Wang, Kai-Sung; Lu, Chi-Yuan; Chang, Shih-Hsien


    This study selected common plant growth regulators (Atonik, Cytokinin, Ethephon, Gibberellic acid and Paclobutrazol) to investigate their biological toxicity to the waters of the important biological indicator Daphnia magna. The methods used in this study included traditional neonate acute toxicity test, new Daphnia embryo toxicity test, and teratogenic embryo test. The study concluded that the acute toxicity of the five PGRs to Daphnia neonate had EC(50) value range of 1.9-130.5 mg l(-1), while acute toxicity of PGRs on Daphnia embryo had EC(50) value range of 0.2-125 mg l(-1); the Daphnia embryos' LOEC values (0.05-48 mg l(-1)) for the five PGRs were lower than embryo EC(50) values. The toxic ratios of 48 h EC(50) (neonate)/48 h LOEC (embryo) for 5 PGRs were 19-512 times. The study found that teratogenic effects of Paclobutrazol and Cytokinin induced in embryo were higher than those of most other PGRs. Microscopic observation of the teratogenic effects showed that all 5 PGRs induced malformations of the second antenna, rostrum, Malpighian tube, sensory bristles, and tail spine as well as function loss and death.

  18. Increase of the processing capacity through modification and enlargement of the assets areas preparation and waste water purification in the fermentation plant Kirchstockach; Durchsatzsteigerung der Vergaerungsanlage Kirchstockach durch Umbau und Erweiterung der Anlagenbereiche Aufbereitung und Prozesswasserreinigung

    Energy Technology Data Exchange (ETDEWEB)

    Kirschenhofer, M. [LRA Muenchen (Germany). Tiefbau, Verkehrsplanung, Abfallwirtschaft; Kroner, T. [ia GmbH - Wissensmanagement und Ingenieurleistungen, Muenchen (Germany). Bereich Kommunale Abfallwirtschaft und Energie; Niefnecker, U. [M. Ganser GmbH und Co. Entsorgungsbetriebe KG, Brunnthal/Kirchstockach (Germany)


    At the fermentation plant Kirchstockach the operations for the rectification of deficiencies and process optimisations were completed in 2004. Now process results of 2005 show the success of the performed actions. In the asset area of preparations the existing rake discharge system was removed and the use of the new discharge reservoir with a drainage coil conveyor system minimises deadlock times and rises preparations throughput. With the new set-up of the light material presses the process procedure was optimised, too. The installation of the new process water reservoir was conditional on the non-uniform hydraulic load of the waste-water purification, which results from the operation of the facility. With the higher buffer capacity, realised by the new process water reservoir, a uniform hydraulic load of the purification system and an optimised process control was implemented. With the optimised performance of the wastewater purification wastewater thresholds are guaranteed now and it is possible to realise the increased throughput of the preparation in the complete system of the fermentation plant Kirchstockach. (orig.)

  19. Purification and partial characterization of a peroxidase from plant cell cultures of Cassia didymobotrya and biotransformation studies. (United States)

    Vitali, A; Botta, B; Delle Monache, G; Zappitelli, S; Ricciardi, P; Melino, S; Petruzzelli, R; Giardina, B


    An acidic peroxidase (EC produced by cell suspension cultures of Cassia didymobotrya (wild senna) was purified from culture medium collected on the 29th day. The enzyme was shown to be a glycoprotein with a pI of 3.5, a molecular mass of approx. 43 kDa by SDS/PAGE and 50 kDa by gel filtration. The N-terminal sequence was very similar to those of other plant peroxidases. The peroxidase was characterized by a high specificity towards coniferyl alcohol and other natural phenolics such as guaiacol and ferulic and caffeic acids. These findings suggest that the enzyme is involved in lignification processes of the cell wall. Moreover, the enzyme was able to catalyse the oxidation of 4,3',4'-trihydroxychalcone and 4, 3',4'-trihydroxy-3-methoxychalcone to the corresponding 3, 3'-biflavanones, as mixtures of racemic and meso forms.

  20. Purification and characterization of a thiol amylase over produced by a non-cereal non-leguminous plant, Tinospora cordifolia. (United States)

    Mukherjee, Abhishek; Ghosh, Anil K; Sengupta, Subhabrata


    A 43kDa α-amylase was purified from Tinospora cordifolia by glycogen precipitation, ammonium sulfate precipitation, gel filtration chromatography, and HPGPLC. The enzyme was optimally active in pH 6.0 at 60°C and had specific activity of 546.2U/mg of protein. Activity was stable in the pH range of 4-7 and at temperatures up to 60°C. PCMB, iodoacetic acid, iodoacetamide, DTNB, and heavy metal ions Hg(2+)>Ag(+)>Cd(2+) inhibited enzyme activity while Ca(2+) improved both activity and thermostability. The enzyme was a thiol amylase (3 SH group/mole) and DTNB inhibition of activity was released by cysteine. N-terminal sequence of the enzyme had poor similarity (12-24%) with those of plant and microbial amylases. The enzyme was equally active on soluble starch and amylopectin and released maltose as the major end product.

  1. Anti-mycobacterial screening of five Indian medicinal plants and partial purification of active extracts of Cassia sophera and Urtica dioica

    Institute of Scientific and Technical Information of China (English)

    Rambir Singh; Shariq Hussain; Rajesh Verma; Poonam Sharma


    Objective: To find out the anti-mycobacterial potential of Cassia sophera (C. sophera), Urticadioica (U. dioica), Momordica dioica, Tribulus terrestris and Coccinia indica plants against multi-drug resistant (MDR) strain of Mycobacterium tuberculosis (M. tuberculosis). Methods: Plant materials were extracted successively with solvents of increasing polarity. Solvent extracts were screened for anti-mycobacterial activity against fast growing, non-pathogenic mycobacterium strain, Mycobacterium semegmatis, by disk diffusion method. The active extracts were tested against MDR and clinical isolates of M. tuberculosis by absolute concentration and proportion methods. The active extracts were subjected to bio-autoassay on TLC followed by silica column chromatography for isolation of potential drug leads. Results: Hexane extract of U. dioica (HEUD) and methanol extract of C. sophera (MECS) produced inhibition zone of 20 mm in disc diffusion assay and MIC of 250 and 125 μg/mL respectively in broth dilution assay against Mycobacteriumsemegmatis. Semipurified fraction F2 from MECS produced 86% inhibition against clinical isolate and 60% inhibition against MDR strain of M. tuberculosis. F18 from HEUD produced 81% inhibition against clinical isolate and 60% inhibition against MDR strain of M. tuberculosis. Phytochemical analysis indicated that anti-mycobacterial activity of MECS may be due to presence of alkaloids or flavonoids and that of HEUD due to terpenoids. Conclusions: C. sophera and U. dioica plant extracts exhibited promising anti-mycobacterial activity against MDR strain of M. tuberculosis. This is the first report of anti-mycobacterial activity form C. sophera. This study showed possibility of purifying novel anti-mycobacterial compound(s) from C. sophera and U. dioica.

  2. Purification and characterization of native and recombinant SaPIN2a, a plant sieve element-localized proteinase inhibitor. (United States)

    Wang, Zhen-Yu; Ding, Ling-Wen; Ge, Zhi-Juan; Wang, Zhaoyu; Wang, Fanghai; Li, Ning; Xu, Zeng-Fu


    SaPIN2a encodes a proteinase inhibitor in nightshade (Solanum americanum), which is specifically localized to the enucleate sieve elements. It has been proposed to play an important role in phloem development by regulating proteolysis in sieve elements. In this study, we purified and characterized native SaPIN2a from nightshade stems and recombinant SaPIN2a expressed in Escherichia coli. Purified native SaPIN2a was found as a charge isomer family of homodimers, and was weakly glycosylated. Native SaPIN2a significantly inhibited serine proteinases such as trypsin, chymotrypsin, and subtilisin, with the most potent inhibitory activity on subtilisin. It did not inhibit cysteine proteinase papain and aspartic proteinase cathepsin D. Recombinant SaPIN2a had a strong inhibitory effect on chymotrypsin, but its inhibitory activities toward trypsin and especially toward subtilisin were greatly reduced. In addition, native SaPIN2a can effectively inhibit midgut trypsin-like activities from Trichoplusia ni and Spodoptera litura larvae, suggesting a potential for the production of insect-resistant transgenic plants.

  3. Purification and Partial Characterization of β-Glucosidase from Fresh Leaves of Tea Plants (Camellia sinensis (L.) O. Kuntze)

    Institute of Scientific and Technical Information of China (English)

    Ye-Yun LI; Chang-Jun JIANG; Xiao-Chun WAN; Zheng-Zhu ZHANG; Da-Xiang LI


    β-Glucosidases are important in the formation of floral tea aroma and the development of resistance to pathogens and herbivores in tea plants. A novel β-glucosidase was purified 117-fold to homogeneity,with a yield of 1.26%, from tea leaves by chilled acetone and ammonium sulfate precipitation, ion exchange chromatography (CM-Sephadex C-50) and fast protein liquid chromatography (FPLC; Superdex 75, Resource S). The enzyme was a monomeric protein with specific activity of 2.57 U/mg. The molecular mass of the enzyme was estimated to be about 41 kDa and 34 kDa by SDS-PAGE and FPLC gel filtration on Superdex 200, respectively. The enzyme showed optimum activity at 50 ℃ and was stable at temperatures lower than 40 ℃. It was active between pH 4.0 and pH 7.0, with an optimum activity at pH 5.5, and was fairly stable from pH 4.5 to pH 8.0. The enzyme showed maximum activity towards pNPG, low activity towards pNP-Galacto, and no activity towards pNP-Xylo.

  4. Malfunction Diagnosing and Analyzing for the Reciprocating Compressor of Dry Gas Purification Ethylene Plant%干气压缩机运行故障诊断研究

    Institute of Scientific and Technical Information of China (English)

    刘文涛; 郭洋


    结合干气提浓乙烯装置的工艺特点,对装置主流程中关键设备往复式压缩机的运行故障进行详细分析,提出介质携带杂质、液体并具有腐蚀性,以及仪表系统的假信号等是导致压缩机故障频发的主要原因.根据实际运行经验,针对导致故障发生的原因从工艺与设备两方面深入探讨改进措施,在实际运行中已采取的措施及取得的良好效果,并就如何提高机组的运行可靠性做了进一步讨论,为干气装置的压缩机长周期稳定运行提出切实可行的方案.%Combining the analysis of the technology characteristics of the reciprocating compressor for "dry gas purification ethylene plant", a detailed analysis is carried on of the operational failure of key e?quipment in the main flow of the device reciprocating compressors, and the main reasons leading to the frequent failure of compressor are raised,which are the medium that carries impurity,liquid and being corrosive,and the false signal in instrument systenuAccording to the actual operating experience,in view of the cause of the failure, technology and equipment are focused on to improve measures,and in the actual opera-lion good results are achieved,and the situation of how to improve the operation reliability of the unit have been further discussed,providing feasible scheme for the stable and long-term operation of reciprocating compressor of dry gas plant.

  5. Screening of Antioxidant Potential of Selected Barks of Indian Medicinal Plants by Multiple in vitro Assays LI

    Institute of Scientific and Technical Information of China (English)



    Objective To evaluate the antioxidant potential in herbal extract barks of five therapeutically important medicinal plants native to India,i.e.Crataeva nurvala Buch.-Ham.,Buchanania lanzan Spreng.,Aegle marmelos Corr.,Dalbergia sissoo Roxb.ex DC.,and Cedrela toona Roxb.Methods Standardized aqueous alcoholic extracts from the selected barks having different target radicals,such as superoxide radical,nitric oxide,ABTS radical,and peroxidative decomposition of phosphohpids.were prepared and screened bv multiple in vitro assays.These extracts were also tested for total phenolic and tannin content and correlated with antioxidant capacity. Results Tbtal phenolic and tannin contents were found to be the highest in C. nurvala (195 GAE mg/g and 218.3 mg/g CE).SOD mimetic activity was found to be the highest in Crataeva nurvula,although all barks showed activity more than 100 units/mg extract.Lipid peroxidation inhibitory potential was found to be the highest in Crataeva nurvala(83.4% inhibition of MDA formation/10 μg extract),and also showed a comparatively high NO quenching capacity (45.5% per 10 μg extract).The highest NO quenching potential was found in Aegle marmelos(47.3% per 10 μg extract).Cedrela toona showed the lowest LPO inhibitory potential and NO quenching capacity(50.5% and 30.5%,respectively).Buchanania lanzan,a medicinal plant extensively used for inflammatory disorders and Dalbergia sissoo also showed 72.5% and 69.1% LPO inhibitory potential/10 μg extract.Trolox equivalent antioxidant capacity ranged from 0.24 to 0.39 mmol/L TEAC/mg extract,indicating that all the barks tested had ABTS+ radical quenching capacity.Conclusion Bark of Crataeva nurvulahas the highest antioxidant capacity and a positive correlation between antioxidant activity and their plendic content was found.

  6. Study on the Purification Effects of Constructed Wetland Plants in NH4+-N Disposal in Living Wastewater%人工湿地园林植物对生活污水中氨氮净化效果的影响

    Institute of Scientific and Technical Information of China (English)

    鲁敏; 郭振; 李东和


    The study on the species selection of wetland plants and the purification effects of the wetland plants is the key to ecological wastewater treatment technology. The purification effects of different wetland plants in different residence time and their synergy in NH4~-N disposal in living wastewater are studied by adopting the surface flow wetland system which is closest to the natural wetlands, through three repeated randomized block field experimental designs, through the variance analysis of two-factor equilibrium of the randomized block with Origin software, and through multiple comparisons and analysis and the test of significant differences with Minitab software. The results show that within the residence time of 1 day, 3 days and 5 days, the purification effects of different plant species and their combinations in NH4*-N disposal are significantly different, and that the NH/-N purification rate of the combination of Arundo donax, Typha orientalis and Canna generate is highest. Within the residence time of 1 day and 3 days, the NH4+-N purification rate of the combination of Phragmites australis and Arundo donax is lowest. Within the residence time of 5 days, the NH4+-N purification rate of Phragmites australis is lowest. The best plant combination in purifying NH4+-N is that of Arundo donax, Typha orientalis and Canna generalis, followed by that of Arundo donax and Typha orientalis or that of Phragmites australis, Typha orientalis and Canna generalis. The best plant species in purifying NH4+-N is Typha orientalis, followed by Arundo donax.%本实验采取了最接近自然湿地表面流的人工湿地系统,通过3次重复的随机区组田间实验设计,采用Origin软件进行随机区组双因素平衡方差分析,采用Minitab软件进行多重比较分析和差异显著性检验,研究了污水不同停留时间和不同人工湿地植物及其协同作用对生活污水中氨氮(NH4+-N)的净化效果.结果表明:停留时间1d、3d、5d时,不同

  7. Application of pressure assisted forward osmosis for water purification and reuse of reverse osmosis concentrate from a water reclamation plant

    KAUST Repository

    Jamil, Shazad


    The use of forward osmosis (FO) is growing among the researchers for water desalination and wastewater treatment due to use of natural osmotic pressure of draw solute. In this study pressure assisted forward osmosis (PAFO) was used instead of FO to increase the water production rate. In this study a low concentration of draw solution (0.25 M KCl) was applied so that diluted KCl after PAFO operation can directly be used for fertigation. The performance of PAFO was investigated for the treatment of reverse osmosis concentrate (ROC) from a water reclamation plant. The water production in PAFO was increased by 9% and 29% at applied pressure of 2 and 4 bars, respectively, to feed side based on 90 h of experiments. Granular activated carbon (GAC) pretreatment and HCl softening were used to reduce organic fouling and scaling prior to application of PAFO. It reduced total organic carbon (TOC) and total inorganic carbon (TIC) by around 90% and 85%, respectively from untreated ROC. Subsequently, this led to an increase in permeate flux. In addition, GAC pretreatment adsorbed 12 out of 14 organic micropollutants tested from ROC to below detection limit. This application enabled to minimise the ROC volume with a sustainable operation and produced high quality and safe water for discharge or reuse. The draw solution (0.25 M KCl) used in this study was diluted to 0.14 M KCl, which is a suitable concentration (10 kg/m3) for fertigation, due to water transport from feed solution. © 2016 Elsevier B.V.

  8. Purification of novel protein elicitor from Botrytis cinerea that induces disease resistance and drought tolerance in plants. (United States)

    Zhang, Yunhua; Yang, Xiufen; Liu, Quan; Qiu, Dewen; Zhang, Yuliang; Zeng, Hongmei; Yuan, Jingjing; Mao, Jianjun


    PebC1, a novel protein elicitor was isolated and purified from the mycelium of gray mold fungus, Botrytis cinerea strain BC-4-2-2-1. The protein was eluted through HiTrap DEAE FF and RESOURCE Q anion exchange chromatography and displayed as a single band with an apparent molecular weight of 36 kDa on silver staining sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The pI of the purified protein PebC1 was determined by 2-DE and was 4.85. Three peptide segments were obtained by MALDI-TOF. Similarity, the homology matching using protein BLAST search found that two proteins, viz. XP_001593856 and XP_001551609 were having high score and covered sequence of the three peptides. Protein XP_001551609, a deduced protein nascent polypeptide-associated complex alpha-polypeptide, was more authentic because it was from Botryotinia fuckeliana that is better known as its anamorph, B.cinerea and showed 95% homology with the three polypeptides. The full cDNA sequence encoding for pebC1 (Genbank accession number FJ748868) was amplified from B. cinerea and consists of 639bp, which is same as a registered gene of XM_001551559, a nascent polypeptide-associated complex alpha-polypeptide partial mRNA. The gene encode a hypothetical protein speculated from an annotated genomic sequence from B. fuckeliana B05.10 (NW_001814507) and there is no publication about the gene. The PebC1 protein significantly promoted wheat seedling growth with an optimum protein concentration of 5 microg/mL. Root systemic activity of wheat with 4-5 leaves increased by 1.29 fold, and the wheat seedling drought resistance integrated index increased from 36.53 to 57.08 under two cycles of drought stress after treatment of PebC1. PebC1 protein at the optimum concentration of 10 microg/mL induced 69.19% disease resistance against gray mold fungus in tomato. Furthermore, phenylalanine ammonia-lyase (PAL), peroxides (POD), and polyphenol oxidase (PPO) related to plant resistance metabolism were also

  9. Application of Isolation and Purification in Natural Plant Pigments%现代分离纯化技术在天然植物色素中的应用

    Institute of Scientific and Technical Information of China (English)

    龚金华; 张玥; 张智超; 孟佳俊; 李德海


    Plant pigment is a kind of natural pigment, which has the characteristics of high nutrition and safety. This essay summarized the research advancements in isolation and purification of natural plant pigments, and laid the foundation for the study of natural plant pigment.%植物色素是一类天然色素,具有营养价值高、安全性强等特点,但是植物色素的分离纯化技术仍然是制约天然色素开发的主要因素。本文就国内天然色素的分离纯化研究进展进行了综述并总结,为天然植物色素的研制与开发奠定基础。

  10. OPTIMASI PROSES DEASIDIFIKASI DALAM PEMURNIAN MINYAK SAWIT MERAH SKALA PILOT PLANT [Optimization of Deacidification Process in Red Palm Oil Purification on Pilot Plant Scale

    Directory of Open Access Journals (Sweden)

    I Wayan Rai Widarta1*


    Full Text Available Deacidification is one of the steps in palm oil refining process which aims to separate free fatty acids formed during post-harvest handling. It is carried out using alkali solution such as NaOH (sodium hydroxide. Carotenoids in palm oil are affected by this step. Therefore, deacidification has to be controlled to minimize the destruction of carotenoids during processing. The objective of this research was to improve deacidification process in pilot plant scale so that the process can produce lower level of free fatty acids (FFA and higher recovery of carotene in high yield neutralized red palm oil (NRPO. Characterization of physical and chemical properties of crude palm oil (CPO such as moisture content, FFA and carotene contents, saponification number, iodine value, peroxide value, and color were determined before processing. Degumming was performed before deacidification process. The 17.5% excess of NaOH was obtained from the pilot plant scale deacidification trial. The optimization of deacidification time and temperature was carried out by using central composite design (CCD. Response surface method (RSM was used to observe the influence of treatments on the FFA level reduction, carotene recovery, and NRPO yield. The result showed that the optimum deacidification condition was at 61 ± 2°C in 26 minutes, and at the 16°Be NaOH strength with 17.5% excess of NaOH. In this optimum condition, the process achieved 96.35% of FFA reduction, 87.30% of carotene recovery, and 90.16% of NRPO yield.

  11. 应用负高压脉冲技术提高植物空气净化能力的探讨%Application of Negative High-voltage Pulse Technology to Improve Air Purification Ability of Plants

    Institute of Scientific and Technical Information of China (English)

    杨运经; 习岗; 刘锴; 张晓辉


    现代城市中的室内空气污染日益严重,室内空气污染的净化与治理已成为社会发展中迫切需要解决的关键科技问题之一.为此,根据植物电信号的低频特征与电共振原理,利用极低频负高压脉冲连续刺激盆栽吊兰植物的根际土壤,使吊兰叶片产生了显著的空气负离子倍增效应.在此基础上依据评价空气质量的常用指标:单极系数q、安培空气质量评价系数CI、空气负离子系数P和森林空气离子评价指数FCI,对应用负高压脉冲技术提高植物的空气净化能力进行了计算与分析.结果表明,极低频负高压脉冲电场可以有效地提高植物的空气净化能力,提出了极低频负高压脉冲电场介导的植物空气净化新技术.%Purification of indoor air pollution has become one of the key technologies of social development. An extremely low frequency high voltage pulse generator was designed according to the characteristics of plants electrical signals and the principle of electrical resonance. The soil of potted broadleaf bracket-plant (Chlorophytum como-sum) was continuously stimulated by negative high voltage pulses with very low frequency, the high concentration of negative air ions appeared around the plant leaves, and the multiplier effect of negative air ion from leave of broadleaf bracket-plant was produced. Then, the ability of plant air purification was calculated and analyzed according to common evaluation indicators of air quality such as single polar factor q, air quality assessment coefficient CI, negative ion air coefficient P and the forest air ion evaluation index FCI. The results show that the extremely low-frequency negative high voltage pulse can enhance the ability of plant to air purification. A new technology of indoor air pollution control and restoration based on negative high-voltage pulse technology is proposed.

  12. Quantitative real-time PCR assay for rapid detection of plant and human pathogenic Macrophomina phaseolina from field and environmental samples. (United States)

    Babu, Bandamaravuri Kishore; Mesapogu, Sukumar; Sharma, Anu; Somasani, Saida Reddy; Arora, Dilip K


    A real-time qPCR assay was developed to detect and quantify Macrophomina phaseolina abundance in rhizosphere soil and plant tissue. Both TaqMan and SYBR green techniques were targeted on ~ 1 kb sequence characterized amplified region (SCAR) of M. phaseolina and two sets of specific primers were designed for SYBR green (MpSyK) and TaqMan (MpTqK) assays. No cross-hybridization and no fluorescent signal exceeding the baseline threshold was observed in TaqMan and SYBR green assays, respectively. The minimum detection limit or sensitivity of TaqMan assay was 30 fg/μL of M. phaseolina DNA and limit of quantification of M. phaseolina viable population was estimated as 0.66 × 10(5) CFU/g soil(-1) equivalent to 10 pg/μL of target DNA. This is the first report which demonstrated real-time qPCR assays with greater specificity and sensitivity to detect M. phaseolina population in soil and plant materials.

  13. Purification of human platelet-derived growth factor

    Energy Technology Data Exchange (ETDEWEB)

    Raines, E.W.; Ross, R.


    The paper describes a method for purification of human platelet-derived growth factor (PDGF) from outdated platelet-rich plasma (PRP) using commonly available laboratory reagents and yielding a mitogen purified 800,000-fold over the starting material. (/sup 3/H)thymidine incorporation into DNA of cultured cells responsive to PDGF represents the most readily available method to follow its purification and define the biological activity of a purified preparation. Other assays to quantitate PDGF include radioreceptor assay and radioimmunoassay.

  14. Development of a photosensitive, high-throughput chip-based superoxide dismutase (SOD) assay to explore the radioprotective activity of herbal plants. (United States)

    Naoghare, Pravin K; Kwon, Ho Taik; Song, Joon Myong


    Appropriate pharmacological interventions and modalities are needed to protect humans against the deleterious effects of ionizing radiation. We disclose a rapid chip-based approach to elucidate the radioprotective/antioxidant potential of herbal plants using a photodiode array (PDA) microchip system. Red light absorption property of nitroblue tetrazolium (NBT) formazan was applied to chip-based superoxide dismutase (SOD) activity measurements of six herbal plant extracts in a high-throughput manner. SOD activities obtained via gel-based assays were in line with the data obtained through the chip-based assay and hence validated our approach. Compared to amifostine, all the tested herbal plant extracts, except apricot kernel, demonstrated greater radioprotective properties. Among the tested herbal extracts, pueraria root showed the highest antioxidant/radioprotective activity and can be considered a preferred radioprotector candidate. Low standard deviations and high statistical confidence obtained during the assay prove the sensitivity and consistency of this approach. The developed approach has several advantages (simplicity, rapidness and portability) over existing methods and can be applied to high-throughput screening of the radioprotective properties of various unexplored plants species.

  15. Multiplex Real-Time qPCR Assay for Simultaneous and Sensitive Detection of Phytoplasmas in Sesame Plants and Insect Vectors.

    Directory of Open Access Journals (Sweden)

    Cengiz Ikten

    Full Text Available Phyllody, a destructive and economically important disease worldwide caused by phytoplasma infections, is characterized by the abnormal development of floral structures into stunted leafy parts and contributes to serious losses in crop plants, including sesame (Sesamum indicum L.. Accurate identification, differentiation, and quantification of phyllody-causing phytoplasmas are essential for effective management of this plant disease and for selection of resistant sesame varieties. In this study, a diagnostic multiplex qPCR assay was developed using TaqMan® chemistry based on detection of the 16S ribosomal RNA gene of phytoplasmas and the 18S ribosomal gene of sesame. Phytoplasma and sesame specific primers and probes labeled with different fluorescent dyes were used for simultaneous amplification of 16SrII and 16SrIX phytoplasmas in a single tube. The multiplex real-time qPCR assay allowed accurate detection, differentiation, and quantification of 16SrII and 16SrIX groups in 109 sesame plant and 92 insect vector samples tested. The assay was found to have a detection sensitivity of 1.8 x 102 and 1.6 x 102 DNA copies for absolute quantification of 16SrII and 16SrIX group phytoplasmas, respectively. Relative quantification was effective and reliable for determination of phyllody phytoplasma DNA amounts normalized to sesame DNA in infected plant tissues. The development of this qPCR assay provides a method for the rapid measurement of infection loads to identify resistance levels of sesame genotypes against phyllody phytoplasma disease.

  16. Comparing the sensitivity of two in vitro assays to evaluate the anthelmintic activity of tropical tannin rich plant extracts against Haemonchus contortus. (United States)

    Alonso-Díaz, M A; Torres-Acosta, J F J; Sandoval-Castro, C A; Hoste, H


    The present trial aimed at comparing the sensitivity of two in vitro methods, i.e. the larval migration inhibition assay (LMIA) and the larval exsheathment inhibition assay (LEIA), to evaluate the anthelmintic (AH) properties of tannin-rich plant extracts against Haemonchus contortus infective larvae. The two assays were applied on the same batch of H. contortus infective larvae exposed to water/acetonic extracts obtained from four tropical plants with different tannin contents: Acacia gaumeri, Brosimum alicastrum, Havardia albicans and Leucaena leucocephala. Increasing concentrations (0, 75, 150, 300, 600, 1200 μg/ml PBS) of lyophilized extracts were used in both in vitro assays. A general lineal model test was used to determine the dose-effect in the LMIA or the difference in the percentage of exsheathed larvae between the respective control and treated groups. The LMIA showed a dose-dependent AH effect for H. albicans (Palicastrum. In contrast, the exsheathment process was significantly affected by all doses of H. albicans and A. gaumeri extracts and a significant dose-dependent effect was found for B. alicastrum and L. leucocephala. Calculation of lethal dose (LD) was possible with LEIA using B. alicastrum and L. leucocephala but not with H. albicans and A. gaumeri as the lowest tested concentration was achieving more than 50% inhibition. Calculation of LD with the LMIA results was not feasible. These results suggest that tannin-rich plant extracts are more potent inhibitors of the exsheathment of H. contortus L(3) larvae than their motility. This information underlines the difference of sensitivity between methodological procedures to evaluate the AH properties of plant extracts on the same nematode stage.

  17. Purification of the major outer membrane protein of Azospirillum brasilense, its affinity to plant roots, and its involvement in cell aggregation. (United States)

    Burdman, S; Dulguerova, G; Okon, Y; Jurkevitch, E


    The major outer membrane protein (MOMP) of the nitrogen-fixing rhizobacterium Azospirillum brasilense strain Cd was purified and isolated by gel filtration, and antiserum against this protein was obtained. A screening of the binding of outer membrane proteins (OMPs) of A. brasilense to membrane-immobilized root extracts of various plant species revealed different affinities for the MOMP, with a stronger adhesion to extracts of cereals in comparison with legumes and tomatoes. Moreover, this protein was shown to bind to roots of different cereal seedlings in an in vitro adhesion assay. Incubation of A. brasilense cells with MOMP-antiserum led to fast agglutination, indicating that the MOMP is a surface-exposed protein. Cells incubated with Fab fragments obtained from purified MOMP-antiserum immunoglobulin G exhibited significant inhibition of bacterial aggregation as compared with controls. Bacteria preincubated with Fab fragments showed weaker adhesion to corn roots in comparison to controls without Fab fragments. These findings suggest that the A. brasilense MOMP acts as an adhesin involved in root adsorption and cell aggregation of this bacterium.

  18. Development of an efficient transformation method by Agrobacterium tumefaciens and high throughput spray assay to identify transgenic plants for woodland strawberry (Fragaria vesca) using NPTII selection. (United States)

    Pantazis, Christopher J; Fisk, Sarah; Mills, Kerri; Flinn, Barry S; Shulaev, Vladimir; Veilleux, Richard E; Dan, Yinghui


    KEY MESSAGE : We developed an efficient Agrobacterium -mediated transformation method using an Ac/Ds transposon tagging construct for F. vesca and high throughput paromomycin spray assay to identify its transformants for strawberry functional genomics. Genomic resources for Rosaceae species are now readily available, including the Fragaria vesca genome, EST sequences, markers, linkage maps, and physical maps. The Rosaceae Genomic Executive Committee has promoted strawberry as a translational genomics model due to its unique biological features and transformability for fruit trait improvement. Our overall research goal is to use functional genomic and metabolic approaches to pursue high throughput gene discovery in the diploid woodland strawberry. F. vesca offers several advantages of a fleshy fruit typical of most fruit crops, short life cycle (seed to seed in 12-16 weeks), small genome size (206 Mbb/C), small plant size, self-compatibility, and many seeds per plant. We have developed an efficient Agrobacterium tumefaciens-mediated strawberry transformation method using kanamycin selection, and high throughput paromomycin spray assay to efficiently identify transgenic strawberry plants. Using our kanamycin transformation method, we were able to produce up to 98 independent kanamycin resistant insertional mutant lines using a T-DNA construct carrying an Ac/Ds transposon Launchpad system from a single transformation experiment involving inoculation of 22 leaf explants of F. vesca accession 551572 within approx. 11 weeks (from inoculation to soil). Transgenic plants with 1-2 copies of a transgene were confirmed by Southern blot analysis. Using our paromomycin spray assay, transgenic F. vesca plants were rapidly identified within 10 days after spraying.


    Venkatesh, Uday; Javarasetty, Chethan; Murari, Satish Kumar


    Wedelia trilobata (L.) Hitch (WT), commonly known as yellow dots or creeping daisy, is a shrub possessing potent biological activities, and is traditionally used a medicinal plant in Ayurveda, Siddha and Unani systems of medicines, and it has also been tried against leukemia cell line MEG- 01. In the present study, purification and screening of the plant was done for bioactive compounds in methanolic extract of WT for apoptotic and anti-leukemia activity. The methanolic extract of WT was initially purified through thin layer chromatography (TLC) and screened for the apoptotic and anti-leukemia activities. The positive band of TLC was subjected to silica gel column chromatography for further purification and the fractions obtained from it were screened again for anti-leukemia activity through thymidine uptake assay and apoptotic activity by DNA fragmentation, nuclear staining and flow cytometry assays. The fraction with positive result was subjected to HPLC for analysis of bioactive components. Out of many combinations of solvents, the methanol and dichloromethane combination in the ratio 6:4 has revealed two bands in TLC, among which the second band showed positive results for apoptotic and anti-leukemic activities. Further purification of second band through silica gel chromatography gave five fractions in which the 3(rd) fraction gave positive results and it shows single peak during compositional analysis through HPLC. The single peak revealed through HPLC indicates the presence of pure compound with apoptotic and anti-leukemia activities encouraging for further structural analysis.

  20. 串联亲和纯化技术及其在植物蛋白质组研究中的应用%Tandem affinity purification technology and its applications in plant proteomic research

    Institute of Scientific and Technical Information of China (English)

    王丰青; 吴为人; 陈新建; 李娟; 周艳; 张重义


    Tandem affinity purification (TAP) is a novel technology for protein complex purification,which has been extensively utilized for the identification or verification of protein-protein interactions and the construction of protein interaction networks.Its basic principle is that the TAP tag is fused to the target protein of interest and then the TAP-tagged protein together with any associated proteins are isolated from the host organism in two sequential affinity purification steps.Initially,TAP technology was mainly used in animals and microbes.But in recent years,more and more applications of TAP technology in plants have been reported with the emergence of new TAP tags and optimized methods,providing an efficient tool for plant proteomics.This paper briefly introduced the application of the TAP method on plants,and analyzed limiting factors in its application.%串联亲和纯化(TAP)是一项新颖的蛋白质复合物纯化技术,已被广泛应用于鉴定或验证蛋白质间的相互作用以及建立蛋白质互作网络,其基本原理是,将TAP标签融合到目标蛋白上,然后经过两步亲和纯化,获得融合蛋白及其结构关联蛋白.TAP技术最初主要在动物和微生物中应用.近几年来,随着新型标签的出现和方法的改进,TAP技术在植物中的应用逐渐增多,为植物蛋白质组学的研究提供了一个有效的手段.简要介绍TAP方法在植物中的应用情况,并对TAP方法应用的限制因素进行分析.

  1. Logic control of molecular sieve purification system of 65000m3· h-1 air separation plant%65000m3·h-1空分设备分子筛纯化系统逻辑控制

    Institute of Scientific and Technical Information of China (English)



    The structure and technical process of molecular sieve purification system of 65000m3/ an air separation plant of Hebi coal and electeicity 600 ktpa methanol project are briefed, automatic valve closing / opening logical control of molecular sieve purification system, automatic / manual operation sequence control program,implementation of the sequence control program and optimized reform of the sequence control configuration program of the molecular sieve absorber in accordance with site operation conditions are described.%简介鹤壁煤电股份有限公司化工分公司空分厂65000m3·h-1空分设备分子筛纯化系统结构和工艺流程,介绍分子筛纯化系统阀门自动开关控制逻辑、顺控程序自动/手动运行、顺控程序执行过程,以及根据现场生产运行实际情况,对分子筛吸附器顺控组态程序进行的优化改进.

  2. Contribution to the optimization of the chemical and radiochemical purification of pressurized water nuclear power plants primary coolant; Contribution a l'optimisation de la purification chimique et radiochimique du fluide primaire des centrales nucleaires a eau sous pression

    Energy Technology Data Exchange (ETDEWEB)

    Elain, L


    The primary coolant of pressurised water reactors is permanently purified thanks to a device, composed of filters and the demineralizers furnished with ion exchange resins (IER), located in the chemical and volume control system (CVCS). The study of the retention mechanisms of the radio-contaminants by the IER implies, initially, to know the speciation of the primary coolant percolant through the demineralizers. Calculations of theoretical speciation of the primary coolant were carried out on the basis of known composition of the primary coolant and thanks to the use of an adapted chemical speciation code. A complementary study, dedicated to silver behaviour, considered badly extracted, suggests metallic aggregates existence generated by the radiolytic reduction of the Ag{sup +} ions. An analysis of the purification curves of the elements Ni, Fe, Co, Cr, Mn, Sb and their principal radionuclides, relating to the cold shutdown of Fessenheim 1-cycle 20 and Tricastin 2-cycle 21, was carried out, in the light of a model based on the concept of a coupling well term - source term. Then, a thermodynamic modelling of ion exchange phenomena in column was established. The formation of the permutation front and the enrichment zones planned was validated by frontal analysis experiments of synthetic fluids (mixtures of Ni(B(OH){sub 4}){sub 2}, LiB(OH){sub 4} and AgB(OH){sub 4} in medium B(OH){sub 3})), and of real fluid during the putting into service of the device mini-CVCS at the time of Tricastin 2 cold shutdown. New tools are thus proposed, opening the way with an optimised management of demineralizers and a more complete interpretation of the available experience feedback. (author)

  3. VAPA, an innovative "virus-acquisition phenotyping assay" opens new horizons in research into the vector-transmission of plant viruses.

    Directory of Open Access Journals (Sweden)

    Alexandre Martinière

    Full Text Available Host-to-host transmission--a key step in plant virus infection cycles--is ensured predominantly by vectors, especially aphids and related insects. A deeper understanding of the mechanisms of virus acquisition, which is critical to vector-transmission, might help to design future virus control strategies, because any newly discovered molecular or cellular process is a potential target for hampering viral spread within host populations. With this aim in mind, an aphid membrane-feeding assay was developed where aphids transmitted two non-circulative viruses [cauliflower mosaic virus (CaMV and turnip mosaic virus] from infected protoplasts. In this assay, virus acquisition occurs exclusively from living cells. Most interestingly, we also show that CaMV is less efficiently transmitted by aphids in the presence of oryzalin--a microtubule-depolymerising drug. The example presented here demonstrates that our technically simple "virus-acquisition phenotyping assay" (VAPA provides a first opportunity to implement correlative studies relating the physiological state of infected plant cells to vector-transmission efficiency.

  4. VAPA, an innovative "virus-acquisition phenotyping assay" opens new horizons in research into the vector-transmission of plant viruses. (United States)

    Martinière, Alexandre; Macia, Jean-Luc; Bagnolini, Guillaume; Jridi, Chiraz; Bak, Aurélie; Blanc, Stéphane; Drucker, Martin


    Host-to-host transmission--a key step in plant virus infection cycles--is ensured predominantly by vectors, especially aphids and related insects. A deeper understanding of the mechanisms of virus acquisition, which is critical to vector-transmission, might help to design future virus control strategies, because any newly discovered molecular or cellular process is a potential target for hampering viral spread within host populations. With this aim in mind, an aphid membrane-feeding assay was developed where aphids transmitted two non-circulative viruses [cauliflower mosaic virus (CaMV) and turnip mosaic virus] from infected protoplasts. In this assay, virus acquisition occurs exclusively from living cells. Most interestingly, we also show that CaMV is less efficiently transmitted by aphids in the presence of oryzalin--a microtubule-depolymerising drug. The example presented here demonstrates that our technically simple "virus-acquisition phenotyping assay" (VAPA) provides a first opportunity to implement correlative studies relating the physiological state of infected plant cells to vector-transmission efficiency.

  5. Screening of antiviral activities in medicinal plants extracts against dengue virus using dengue NS2B-NS3 protease assay. (United States)

    Rothan, H A; Zulqarnain, M; Ammar, Y A; Tan, E C; Rahman, N A; Yusof, R


    Dengue virus infects millions of people worldwide and there is no vaccine or anti-dengue therapeutic available. Screening large numbers of medicinal plants for anti-dengue activities is an alternative strategy in order to find the potent therapeutic compounds. Therefore, this study was designed to identify anti-dengue activities in nineteen medicinal plant extracts that are used in traditional medicine. Local medicinal plants Vernonia cinerea, Hemigraphis reptans, Hedyotis auricularia, Laurentia longiflora, Tridax procumbers and Senna angustifolia were used in this study. The highest inhibitory activates against dengue NS2B-NS3pro was observed in ethanolic extract of S. angustifolia leaves, methanolic extract of V. cinerea leaves and ethanol extract of T. procumbens stems. These findings were further verified by in vitro viral inhibition assay. Methanolic extract of V. cinerea leaves, ethanol extract of T. procumbens stems and at less extent ethanolic extract of S. angustifolia leaves were able to maintain the normal morphology of DENV2-infected Vero cells without causing much cytopathic effects (CPE). The percentage of viral inhibition of V. cinerea and T. procumbens extracts were significantly higher than S. angustifolia extract as measured by plaque formation assay and RT-qPCR. In conclusion, The outcome of this study showed that the methanolic extract of V. cinerea leaves and ethanol extract of T. procumbens stems possessed high inhibitory activates against dengue virus that worth more investigation.

  6. Purification of contaminated groundwater by membrane technology

    Energy Technology Data Exchange (ETDEWEB)

    Youn, In Soo; Chung, Chin Ki; Kim, Byoung Gon [Korea Institute of Geology Mining and Materials, Taejon (Korea, Republic of)


    The objective of this study is to apply the membrane separation technology to the purification of contaminated ground water in Korea. Under this scope, the purification was aimed to the drinking water level. The scale of the membrane system was chosen to a small filtration plant for local clean water supplies and/or heavy purifiers for buildings and public uses. The actual conditions of ground water contamination in Korea was surveyed to determine the major components to remove under the drinking water requirements. To set up a hybrid process with membrane methods, conventional purification methods were also investigated for the comparison purpose. The research results are summarized as follows : 1) Contamination of the groundwater in Korea has been found to be widespread across the country. The major contaminant were nitrate, bacteria, and organic chlorides. Some solvents and heavy metals are also supposed to exist in the ground water of industrial complexes, cities, and abandoned mines. 2) The purification methods currently used in public filtration plants appear not to be enough for new contaminants from recent industrial expanding. The advanced purification technologies generally adopted for this problem have been found to be unsuitable due to their very complicated design and operation, and lack of confidence in the purification performance. 3) The reverse osmosis tested with FilmTec FT30 membrane was found to remove nitrate ions in water with over 90 % efficiency. 4) The suitable membrane process for the contaminated groundwater in Korea has been found to be the treatments composed of activated carbon, microfiltration, reverse osmosis or ultrafiltration, and disinfection. The activated carbon treatment could be omitted for the water of low organic contaminants. The microfiltration and the reverse osmosis treatments stand for the conventional methods of filtration plants and the advanced methods for hardly removable components, respectively. It is recommended

  7. Simultaneous quantification of five bacterial and plant toxins from complex matrices using a multiplexed fluorescent magnetic suspension assay. (United States)

    Pauly, Diana; Kirchner, Sebastian; Stoermann, Britta; Schreiber, Tanja; Kaulfuss, Stefan; Schade, Rüdiger; Zbinden, Reto; Avondet, Marc-André; Dorner, Martin B; Dorner, Brigitte G


    Proteotoxins such as ricin, abrin, botulinum neurotoxins type A and B (BoNT/A, BoNT/B) and staphylococcal enterotoxin B (SEB) are regarded as potential biological warfare agents which could be used for bioterrorism attacks on the food chain. In this study we used a novel immunisation strategy to generate high-affinity monoclonal and polyclonal antibodies against native ricin, BoNT/A, and BoNT/B. The antibodies were used along with antibodies against SEB and abrin to establish a highly sensitive magnetic and fluorescent multiplex bead array with excellent sensitivities between 2 ng/L and 546 ng/L from a minimal sample volume of 50 microL. The assay was validated using 20 different related analytes and the assay precision was determined. Advancing the existing bead array technology, the novel magnetic and fluorescent microbeads proved amenable to enrichment procedures, by further increasing sensitivity to 0.3-85 ng/L, starting from a sample volume of 500 microL. Furthermore, the method was successfully applied for the simultaneous identification of the target toxins spiked into complex food matrices like milk, baby food and yoghurt. On the basis of our results, the assay appears to be a good tool for large-scale screening of samples from the food supply chain.

  8. Selection of Streptomyces against soil borne fungal pathogens by a standardized dual culture assay and evaluation of their effects on seed germination and plant growth. (United States)

    Kunova, Andrea; Bonaldi, Maria; Saracchi, Marco; Pizzatti, Cristina; Chen, Xiaoyulong; Cortesi, Paolo


    In the search for new natural resources for crop protection, streptomycetes are gaining interest in agriculture as plant growth promoting bacteria and/or biological control agents. Because of their peculiar life cycle, in which the production of secondary metabolites is synchronized with the development of aerial hyphae and sporulation, the commonly used methods to screen for bacterial antagonists need to be adapted. The dual culture assay was standardized in terms of inoculation timing of Streptomyces antagonist and pathogen, and growth rate of different fungal pathogens. In case of fast-growing fungi, inoculation of the antagonist 2 or 3 days prior to the pathogen resulted in significantly stronger inhibition of mycelium growth. One hundred and thirty Streptomyces strains were evaluated against six destructive soil borne pathogens. The activity of strains varied from broad-spectrum to highly specific inhibition of individual pathogens. All strains inhibited at least one tested pathogen. Three strains, which combined the largest broad-spectrum with the highest inhibition activity, were selected for further characterization with four vegetable species. All of them were able to colonize seed surface of all tested vegetable crops. They mostly improved radicle and hypocotyl growth in vitro, although no statistically significant enhancement of biomass weight was observed in vivo. Occasionally, transient negative effects on germination and plant growth were observed. The adapted dual culture assay allowed us to compare the inhibition of individual Streptomyces strains against six fungal soil borne pathogens. The best selected strains were able to colonize the four vegetable crops and have a potential to be developed into biocontrol products. Although they occasionally negatively influenced plant growth, these effects did not persist during the further development. Additional in vivo studies are needed to confirm their potential as biological control or plant growth

  9. An in vivo root hair assay for determining rates of apoptotic-like programmed cell death in plants

    Directory of Open Access Journals (Sweden)

    Hogg Bridget V


    Full Text Available Abstract In Arabidopsis thaliana we demonstrate that dying root hairs provide an easy and rapid in vivo model for the morphological identification of apoptotic-like programmed cell death (AL-PCD in plants. The model described here is transferable between species, can be used to investigate rates of AL-PCD in response to various treatments and to identify modulation of AL-PCD rates in mutant/transgenic plant lines facilitating rapid screening of mutant populations in order to identify genes involved in AL-PCD regulation.

  10. Necessity of Purification during Bacterial DNA Extraction with Environmental Soils. (United States)

    Lim, Hyun Jeong; Choi, Jung-Hyun; Son, Ahjeong


    Complexity and heterogeneity of soil samples have often implied the inclusion of purification steps in conventional DNA extraction for PCR assays. Unfortunately the purification steps are also time and labor intensive. Therefore the necessity of DNA purification was re-visited and investigated for a variety of environmental soil samples that contained various amounts of PCR inhibitors. Bead beating and centrifugation was used as the baseline (without purification) method for DNA extraction. Its performance was compared with that of conventional DNA extraction kit (with purification). The necessity criteria for DNA purification were established with environmental soil samples. Using lysis conditions at 3000 rpm for 3 minutes with 0.1 mm glass beads, centrifugation time of 10 minutes and 1:10 dilution ratio, the baseline method outperformed conventional DNA extraction on cell seeded sand samples. Further investigation with PCR inhibitors (i.e., humic acids, clay, and magnesium) showed that sand samples containing less than 10 ug/g humic acids and 70% clay may not require purifications. Interestingly, the inhibition pattern of magnesium ion was different from other inhibitors due to the complexation interaction of magnesium ion with DNA fragments. It was concluded that DNA extraction method without purification is suitable for soil samples that have less than 10 ug/g of humic acids, less than 70% clay content and less than 0.01% magnesium ion content.

  11. An Optimized Microplate Assay System for Quantitative Evaluation of Plant Cell Wall Degrading Enzyme Activity of Fungal Culture Extracts (United States)

    Developing enzyme cocktails for cellulosic biomass hydrolysis complementary to current cellulase systems is a critical step needed for economically viable biofuels production. Recent genomic analysis indicates that some plant pathogenic fungi are likely a largely untapped resource in which to prospe...

  12. Assessment of two medicinal plants, Psidium guajava L. and Achillea millefolium L., in in vitro and in vivo assays

    Directory of Open Access Journals (Sweden)

    Teixeira Rosangela de Oliveira


    Full Text Available The use of medicinal plants by the general population is an old and still widespread practice, which makes studies of their genotoxicity essential. Psidium guajava L. and Achillea millefolium L. are examples of plants commonly used in popular medicine. P. guajava L. is indicated for diarrhea and also as an antiseptic, while A. millefolium L. is indicated as an analgesic, antispasmodic, digestive, diuretic, antiseptic, astringent, emollient, wound healer and hemorrhoid medication. The aim of this study was to determine the effects of the infusions of these two plant species on chromosomes and the cell cycle. Leaves from the plants were used to prepare infusions, in the same manner as teas, but at two different concentrations. Allium cepa L. root-tip cells (P. guajava L. - 2.62 and 26.2 mg/mL, and A. millefolium L. - 3.5 and 35.0 mg/mL and Wistar rat bone marrow cells (P. guajava L. - 2.62 and 26.2 mg/100g body weight, and A. millefolium L. - 3.5 and 35.0 mg/100g body weight were used as in vivo plant and animal test systems, respectively. Human peripheral blood lymphocytes (P. guajava L. - 0.262 and 2.62 mg/mL culture medium, and A. millefolium L. - 0.35 and 3.5 mg/mL culture medium were used as in vitro test system. The P. guajava L. infusion at the higher concentration caused a statistically significant inhibition of cellular division in the onion root-tip cells, not observed in onion root-tip cells treated with A. millefolium L. No statistically significant alterations were found, as compared to untreated controls, in either the cell cycle or the number of chromosome alterations, after treatments with either plant, in rat cells or in cultured human lymphocytes. These results regarding the cytotoxicity and mutagenicity of these plants provide valuable information about the safety of using them as therapeutic agents.

  13. One-step purification of phosphinothricin acetyltransferase using reactive dye-affinity chromatography. (United States)

    Wang, Cunxi; Lee, Thomas C; Crowley, Kathleen S; Bell, Erin


    Reactive dye purification is an affinity purification technique offering unique selectivity and high purification potential. Historically, purification of phosphinothricin acetyltransferase (PAT) has involved several steps of precipitation and column chromatography. Here, we describe a novel purification method that is simple, time-saving, inexpensive, and reproducible. The novel method employs a single chromatography step using a reactive dye resin, Reactive brown 10-agarose. Reactive brown 10 preferentially binds the PAT protein, which can then be specifically released by one of its substrates, acetyl-CoA. Using Reactive brown 10-agarose, PAT protein can be purified to homogeneity from E. coli or plant tissue with high recovery efficiency.

  14. Unsuitability of 2,3,5-Triphenyl-2H-Tetrazolium chloride(TTC) as a viability assay for plant cells in suspention

    Energy Technology Data Exchange (ETDEWEB)

    Kebler, M.; Furusaki, S. [Department of Chemistry and Biotechnology Univ, Tokyo (Japan). Graduate School of Engineering


    The well-known viability assay with 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) is applied to plant cell suspensions. In this paper it is shown that parameters (pH, TTC concentration, incubation time) which are thought to b e only dependent on different cell lines are underlying at least two additional functions (age of the culture and shear stress). Each cell in a different state of activity requires a new set of the parameters mentioned above. Furthermore the time-dependent formazan production courses vary to such an extent that they cannot be used for viability determination. Therefore the usage of TTC as a viability test implies non negligible errors compared to the Evans` Blue staining method which does not involve cell metabolism. The values of the two different methods to determine the viability can differ by more than 50%. The data suggests abandoning the usage of TTC as a quantitative viability assay for plant cell suspensions. 22 refs., 7 figs., 1 tab.

  15. An in vivo assay for elucidating the importance of cytochromes P450 for the ability of a wild mammalian herbivore (Neotoma lepida) to consume toxic plants. (United States)

    Skopec, Michele M; Malenke, Jael R; Halpert, James R; Denise Dearing, M


    An in vivo assay using the cytochrome P450 (P450) suicide inhibitor 1-aminobenzotriazole (ABT) and 24-h food intake was developed to determine the relative importance of P450s in two populations of Neotoma lepida with respect to biotransformation of plant secondary compounds in the animals' natural diets. The efficacy of ABT as a P450 inhibitor was first validated using hypnotic-state assays with and without pretreatment with ABT. Pretreatment with 100 mg/kg ABT by gavage increased hexobarbital sleep times 3-4-fold in both populations, showing effective inhibition of P450s in woodrats. Next, the Great Basin population was fed a terpene-rich juniper diet, and the Mojave population was fed a phenolic-rich creosote diet, with rabbit chow serving as the control diet in each group. Treatment with ABT inhibited food intake in the Great Basin population fed the juniper diet to a greater extent (35%) than the Great Basin population fed the control diet (19%) or the Mojave population fed the creosote diet (16%). The food intake of the Mojave population fed the control diet was not significantly inhibited by ABT. The findings suggest that the biotransformation of terpenes in juniper relies more heavily on P450s than that of phenolics in creosote. This assay provides an inexpensive and noninvasive method to explore the relative importance of P450s in the biotransformation strategies of wild mammalian herbivores.

  16. 含硫天然气净化厂硫磺安全储存的有关探讨%Study on the Safety Storage Design of Sulfur of Sour Natural Gas Purification Plant

    Institute of Scientific and Technical Information of China (English)

    张海林; 向敏; 杨毅


    硫磺是含硫天然气脱硫净化的重要附属产物,硫磺的安全储存对于含硫天然气净化厂的安全高效运行具有重要意义。论文分析了天然气净化厂硫磺燃烧爆炸反应历程,基于实验测试参数,探讨了硫磺粉末爆炸危险特性,深入研究了硫磺粒度、含水量等关键因素对爆炸特性的影响规律及趋势;通过对硫磺燃烧及爆炸规律分析研究,提出了硫磺料仓安全监控关键技术方案,构建了含硫天然气净化厂硫磺储存单元消防设计的总体思路及方法,为含硫天然气净化厂硫磺储存设计及安全管理提供了参考。%Sulfur is an important ancillary product in the process of desulfurization and purification of high-sour natural gas. Experiment Study on sulfur combustion rate was carried out. Characteristic parameters of sulfur explosion under different conditions were tested and analyzed. Key factors influencing sulfur dust combustion and the effect Characteristic were studied. The overall train of thought and method of safety monitoring and fire control design of sulfur storage bin were proposed. The work provided reference for sulfur storage design and safety management of sour natural gas purification plant.

  17. Directly Transforming PCR-Amplified DNA Fragments into Plant Cells Is a Versatile System That Facilitates the Transient Expression Assay (United States)

    Lu, Yuming; Chen, Xi; Wu, Yuxuan; Wang, Yanping; He, Yuqing; Wu, Yan


    A circular plasmid containing a gene coding sequence has been broadly used for studying gene regulation in cells. However, to accommodate a quick screen plasmid construction and preparation can be time consuming. Here we report a PCR amplified dsDNA fragments (PCR-fragments) based transient expression system (PCR-TES) for suiting in the study of gene regulation in plant cells. Instead of transforming plasmids into plant cells, transient expression of PCR-fragments can be applicable. The transformation efficiency and expression property of PCR-fragments are comparable to transformation using plasmids. We analyzed the transformation efficiency in PCR-TES at transcription and protein levels. Our results indicate that the PCR-TES is as versatile as the conventional transformation system using plasmid DNA. Through reconstituting PYR1-mediated ABA signaling pathway in Arabidopsis mesophyll protoplasts, we were not only validating the practicality of PCR-TES but also screening potential candidates of CDPK family members which might be involved in the ABA signaling. Moreover, we determined that phosphorylation of ABF2 by CPK4 could be mediated by ABA-induced PYR1 and ABI1, demonstrating a crucial role of CDPKs in the ABA signaling. In summary, PCR-TES can be applicable to facilitate analyzing gene regulation and for the screen of putative regulatory molecules at the high throughput level in plant cells. PMID:23468926

  18. Directly transforming PCR-amplified DNA fragments into plant cells is a versatile system that facilitates the transient expression assay.

    Directory of Open Access Journals (Sweden)

    Yuming Lu

    Full Text Available A circular plasmid containing a gene coding sequence has been broadly used for studying gene regulation in cells. However, to accommodate a quick screen plasmid construction and preparation can be time consuming. Here we report a PCR amplified dsDNA fragments (PCR-fragments based transient expression system (PCR-TES for suiting in the study of gene regulation in plant cells. Instead of transforming plasmids into plant cells, transient expression of PCR-fragments can be applicable. The transformation efficiency and expression property of PCR-fragments are comparable to transformation using plasmids. We analyzed the transformation efficiency in PCR-TES at transcription and protein levels. Our results indicate that the PCR-TES is as versatile as the conventional transformation system using plasmid DNA. Through reconstituting PYR1-mediated ABA signaling pathway in Arabidopsis mesophyll protoplasts, we were not only validating the practicality of PCR-TES but also screening potential candidates of CDPK family members which might be involved in the ABA signaling. Moreover, we determined that phosphorylation of ABF2 by CPK4 could be mediated by ABA-induced PYR1 and ABI1, demonstrating a crucial role of CDPKs in the ABA signaling. In summary, PCR-TES can be applicable to facilitate analyzing gene regulation and for the screen of putative regulatory molecules at the high throughput level in plant cells.

  19. Purification of tubulin from porcine brain. (United States)

    Gell, Christopher; Friel, Claire T; Borgonovo, Barbara; Drechsel, David N; Hyman, Anthony A; Howard, Jonathon


    Microtubules, polymers of the heterodimeric protein αβ-tubulin, give shape to cells and are the tracks for vesicle transport and chromosome segregation. In vitro assays to study microtubule functions and their regulation by microtubule-associated proteins require the availability of purified αβ-tubulin. In this chapter, we describe the process of purification of heterodimeric αβ-tubulin from porcine brain.

  20. Gas purification process

    Energy Technology Data Exchange (ETDEWEB)

    Hoelter, H.; Gresch, H.; Igelbuescher, H.; Dewert, H.


    To avoid the problems of reheating in a wet process as well as the problems of higher gas supply in a dry process, the invention proposes to separate the raw gas in two component currents, one of which undergoes wet purification while the other is led through a dry purification process. The two component currents are mixed before entering the stack. The dry chemisorption masses added in substoichiometric doses are treated in a milk-of-lime processing stage, after which the reacted and non-reacted chemisorption masses are treated by wet purification and then by oxidation.

  1. The Borexino purification system (United States)

    Benziger, Jay


    Purification of 278 tons of liquid scintillator and 889 tons of buffer shielding for the Borexino solar neutrino detector is performed with a system of combined distillation, water extraction, gas stripping and filtration. The purification system removed K, U and Th by distillation of the pseudocumene solvent and the PPO fluor. Noble gases, Rn, Kr and Ar were removed by gas stripping. Distillation was also employed to remove optical impurities and reduce the attenuation of scintillation light. The success of the purification system has facilitated the first time real time detection of low energy solar neutrinos.

  2. Activity of Medicinal Plant Extracts on Multiplication of Mycobacterium tuberculosis under Reduced Oxygen Conditions Using Intracellular and Axenic Assays

    Directory of Open Access Journals (Sweden)

    Purva D. Bhatter


    Full Text Available Aim. Test the activity of selected medicinal plant extracts on multiplication of Mycobacterium tuberculosis under reduced oxygen concentration which represents nonreplicating conditions. Material and Methods. Acetone, ethanol and aqueous extracts of the plants Acorus calamus L. (rhizome, Ocimum sanctum L. (leaf, Piper nigrum L. (seed, and Pueraria tuberosa DC. (tuber were tested on Mycobacterium tuberculosis H37Rv intracellularly using an epithelial cell (A549 infection model. The extracts found to be active intracellularly were further studied axenically under reducing oxygen concentrations. Results and Conclusions. Intracellular multiplication was inhibited ≥60% by five of the twelve extracts. Amongst these 5 extracts, in axenic culture, P. nigrum (acetone was active under aerobic, microaerophilic, and anaerobic conditions indicating presence of multiple components acting at different levels and P. tuberosa (aqueous showed bactericidal activity under microaerophilic and anaerobic conditions implying the influence of anaerobiosis on its efficacy. P. nigrum (aqueous and A. calamus (aqueous and ethanol extracts were not active under axenic conditions but only inhibited intracellular growth of Mycobacterium tuberculosis, suggesting activation of host defense mechanisms to mediate bacterial killing rather than direct bactericidal activity.

  3. Functional Characterization of a Bidirectional Plant Promoter from Cotton Leaf Curl Burewala Virus Using an Agrobacterium-Mediated Transient Assay

    Directory of Open Access Journals (Sweden)

    Muhammad Aleem Ashraf


    Full Text Available The C1 promoter expressing the AC1 gene, and V1 promoter expressing the AV1 gene are located in opposite orientations in the large intergenic region of the Cotton leaf curl Burewala virus (CLCuBuV genome. Agro-infiltration was used to transiently express putative promoter constructs in Nicotiana tabacum and Gossypium hirsutum leaves, which was monitored by a GUS reporter gene, and revealed that the bidirectional promoter of CLCuBuV transcriptionally regulates both the AC1 and AV1 genes. The CLCuBuV C1 gene promoter showed a strong, consistent transient expression of the reporter gene (GUS in N. tabacum and G. hirsutum leaves and exhibited GUS activity two- to three-fold higher than the CaMV 35S promoter. The CLCuBuV bidirectional gene promoter is a nearly constitutive promoter that contains basic conserved elements. Many cis-regulatory elements (CREs were also analyzed within the bidirectional plant promoters of CLCuBuV and closely related geminiviruses, which may be helpful in understanding the transcriptional regulation of both the virus and host plant.

  4. 盆栽植物对室内甲醛空气污染的净化研究进展%Research Advance in Purification of Formaldehyde-polluted Indoor Air by Potted Plants

    Institute of Scientific and Technical Information of China (English)

    何勤勤; 周俊辉


    Formaldehyde ( FDH) is a main indoor air pollutant , and it can cause serious hazards to human ’ s physical and mental health.Therefore, it is one of today’s hot research subjects that the air pollutants from construction and decoration materials can be absorbed and removed by potted plants .Many researches showed that the indoor FDH concentration could be reduced effec -tively by many potted plants .The purification of FHD maybe included several aspects , such as assimilation by stems or leaves of potted plants , transformation and metabolism by plant cells , and degradation by the rhizospheric microorganisms .There maybe three ways for potted plants to react with FDH pollution:the first one had high absorption but weak resistance to FDH damage , showing obvious hurt external morphology;the second one had weak absorption but strong resistance to FDH damage , revealing normal exter-nal morphology through taking avoidance strategy to protect itself;the third one showed the strongest absorption and transforming a-bility to FDH with more or less hurt responses .The purification mechanism of formaldehyde -polluted indoor air by potted plants still needs for further researches .%甲醛是室内主要的空气污染物,严重影响了人们的身心健康,寻找净化空气污染的高效盆栽植物成为近年来研究的热点。许多研究表明:盆栽植物都能有效地降低室内甲醛的浓度,对甲醛的净化作用可能有通过盆栽植物的茎叶表面吸收、植物细胞转化代谢、根际微生物吸收降解等几个方面。植物对甲醛净化可能存在3种途径或机制:第1种是对甲醛吸收能力强,但抗性弱,外部形态伤害明显;第2种是植物单位干物质吸收甲醛量较少,但采取保护自己的规避策略,保持外部形态的完整;第3种是吸收并转化,这类植物单位干物质甲醛吸收量高,同时外部形态表现基本正常或正常。对盆栽植物净化甲醛空气

  5. Analysis and Comparison of Water Purification Processes and Finished Water Quality for 3 Water Treatment Plants%关于三个水厂净水工艺与供水水质的比较和分析

    Institute of Scientific and Technical Information of China (English)

    岳宇明; 陆茸; 毛丽娜; 沈元静; 何小清


    该文介绍了国内一自来水公司三个水厂的两个原水水质、净水工艺及出厂水水质。结果显示第一水厂的出厂水质较为理想,第二水厂次之,第三水厂为第三。第三水厂由于水源的问题导致出厂氨氮季节性超标,建议采取有效措施改进水源水质,以提高出厂水质。第二水厂需进行工艺改造,实施臭氧活性炭深度处理以进一步提高供水水质。第三水厂一期系统臭氧生物活性炭池置于砂滤池后较二期活性炭滤池置于砂滤池前出水有机物CODMn及TOC略低,但两者基本相近。建议第三水厂采取必要的措施改进水源水质,或再增加一道臭氧生物活性炭工序。%Water plants with two different raw water qualities,purification processes and their effluent water qualities were introduced in this paper. The finished water quality of NO. 1 plant is the best,and the NO. 2 is better. Due to raw water quality problem No. 3 water plant’s finished water ammonia nitrogen hardly meets the standard of GB 5749-2006 seasonally. Effective measures should be taken to improve the effluent water quality of No. 3 water plant No. 2 water plant need technological transformation to implement advanced treatment of O3 actived carbon to improve finished water quality. The organic indexes of effluent water of first-stage system in No. 3 water plant,of which O3 actived carbon filter is located behind the sand filter,is a little better than that of second-stage system, of which O3 actived carbon filter is located before the sand filter. But their finished water qualities are approximately same. Necessary measures to improve raw water quality for No. 3 water plant or to apply an additional ozone BAC process are recommended.

  6. Application of Antibiotics to Plant Protection and Research Progress of Their Isolation and Purification%抗生素在植物保护中的应用及分离纯化研究进展

    Institute of Scientific and Technical Information of China (English)

    王苗苗; 刘峥; 袁帅; 郝再彬


    There are many advantages for the antibiotics which are used to protect plant,such as wonderful environmental compatibility and high selectivity,etc. Instead of chemical pesticides,antibiotics are used to control crops diseases and insects. At the same time,it is an inevitable trend for the modern green agriculture development. Antibiotics are commonly produced by microorganisms in the fermentation process. However,the antibiotics above are low in content with impurities. As a result,they are extremely difficult to be isolated and purified, which restrict their applications to plant protection. In this paper, the types of antibiotics,their application to plant protection,and the research progress of their isolation and purification are summarized.%抗生素用于植物保护具有选择性强、与环境兼容性好等优点,以抗生素代替化学农药来防治植物病虫草害,是现代绿色农业发展的必然趋势.抗生素一般由微生物在发酵过程中产生,含量很低,且发酵液中杂质很多,分离纯化极为困难,制约了抗生素在植物保护方面的应用.就抗生素的种类、在植物保护中的应用及其分离纯化方法的研究进展进行了归纳.

  7. New data on electron-beam purification of wastewater

    Energy Technology Data Exchange (ETDEWEB)

    Pikaev, A.K. E-mail:


    Recent environmental applications of radiation technology, developed in the author's laboratory, are presented in this paper. They are electron-beam and coagulation purification of molasses distillery slops from distillery-produced ethyl alcohol by fermentation of plant materials, electron-beam purification of wastewater from carboxylic acids (for example, formic acid) and removal of petroleum products (diesel fuel, motor oil and residual fuel oil) from water by {gamma}-irradiation.

  8. New data on electron-beam purification of wastewater (United States)

    Pikaev, A. K.


    Recent environmental applications of radiation technology, developed in the author's laboratory, are presented in this paper. They are electron-beam and coagulation purification of molasses distillery slops from distillery-produced ethyl alcohol by fermentation of plant materials, electron-beam purification of wastewater from carboxylic acids (for example, formic acid) and removal of petroleum products (diesel fuel, motor oil and residual fuel oil) from water by γ-irradiation.

  9. Occurrence and genetic diversity of Arcobacter spp. in a spinach-processing plant and evaluation of two Arcobacter-specific quantitative PCR assays. (United States)

    Hausdorf, Lena; Neumann, Maria; Bergmann, Ingo; Sobiella, Kerstin; Mundt, Kerstin; Fröhling, Antje; Schlüter, Oliver; Klocke, Michael


    Some species of the genus Arcobacter are considered to be emerging food pathogens. With respect to recent vegetable-borne outbreaks, the aim of this work was to investigate the occurrence and diversity of Arcobacter within the production chain of a spinach-processing plant by a combination of cultivation and molecular methods. Samples including spinach, water, and surface biofilm were taken over a period of three years from the entire processing line. Ten 16S rRNA (rrs) gene clone libraries were constructed and analysed using amplified rRNA gene restriction analysis (ARDRA). Approximately 1200 clones were studied that resulted in 44 operational taxonomic units (OTUs). Sequences with high similarities to Arcobacter cryaerophilus (13% of clones, 3 OTUs), A. ellisii (4%, 6 OTUs), A. suis (15%, 3 OTUs), and the type strain of A. nitrofigilis (1%, 7 OTUs) were identified. This represents the first report of the detection of the recently described species A. ellisii, A. suis and, in addition, A. venerupis from alternative habitats. A total of 67% of the clones (22 OTUs) could not be assigned to a genus, which indicated the presence of uncharacterised Arcobacter species. For the cultivation-independent detection of Arcobacter, two genus-specific quantitative PCR (qPCR) assays were developed and tested on 15 Arcobacter species. When these assays were applied to samples from the spinach-processing plant, they showed positive results for up to 35% of the samples and supported the conclusion that there is a considerable risk for the transfer of pathogenic Arcobacter species on vegetables, which was also verified by a cultivation approach.

  10. A fluorescence polarization assay to quantify biotin and biotin-binding proteins in whole plant extracts using Alexa-Fluor 594 biocytin. (United States)

    Martin, Harry; Murray, Colleen; Christeller, John; McGhie, Tony


    A high-throughput fluorescence polarization assay has been developed for the detection of biotin and biotin-binding proteins in whole leaf extracts. Various groups are investigating the insecticidal properties of avidin and other biotin-binding proteins expressed in leaves of transgenic plants. The methods commonly used to quantify biotin and avidin in leaf extracts are enzyme-linked immunosorbent assay (ELISA) and Western blotting. Here we describe a homogeneous fluorescence polarization (FP) method that quantifies transgenic avidin in whole leaf extract by the simple addition of the fluorescent avidin ligand Alexa-Fluor 594 biocytin (AFB). The FP assay exploits the fact that AFB excites and emits in regions of the spectrum that are relatively free of background fluorescence in leaf extract. Transgenic leaf avidin can be quantified within 1-2 h by the FP method, in comparison with 1-2 days for ELISA and Western blotting. The FP method can also measure the amount of biotin in control leaves, not expressing avidin. Functional avidin levels of 1.54 microM (26.1 microg/g leaf tissue) were detected in tobacco leaves expressing vacuole-targeted avidin. Control leaves had biotin levels of around 0.74 microM (approximately 0.18 microg/g leaf tissue). Reagent costs are minimal: typically AFB is used at concentrations of 1-10 nM, avidin is used at 1-100 nM, and sample volumes are 20 microL in 384-well microplates.

  11. Screening of agonistic activities against four nuclear receptors in wastewater treatment plants in Japan using a yeast two-hybrid assay

    Institute of Scientific and Technical Information of China (English)

    Daisuke Inoue; Koki Nakama; Kazuko Sawada; Taro Watanabe; Hisae Matsui; Kazunari Sei; Tsuyoshi Nakanishi; Michihiko Ike


    To assess the potential endocrine disruptive effects through multiple nuclear receptors (NRs), especially non-steroidal NRs, in municipal wastewater, we examined the agonistic activities on four NRs (estrogen receptor c, thyroid hormone receptor α, retinoic acid receptor α and retinoid X receptor α) of untreated and treated wastewater from municipal wastewater treatment plants (WWTPs) in Japan using a yeast two-hybrid assay.Investigation of the infiuent and effluent of seven WWTPs revealed that agonistic activities against steroidal and non-steroidal NRs were always detected in the infiuents and partially remained in the effluents.Further investigation of four WWTPs employing conventional activated sludge, pseudo-anoxic-oxic, anoxic-oxic and anaerobic-anoxic-oxic processes revealed that the ability to reduce the agonistic activity against each of the four NRs varies depending on the treatment process.These results indicated that municipal wastewater in Japan commonly contains endocrine disrupting chemicals that exert agonistic activities on steroidal and non-steroidal NRs, and that some of these chemicals are released into the natural aquatic environment.Although the results obtained in yeast assays suggested that measured levels of non-steroidal NR agonists in the effluent of WWTPs were not likely to cause any biological effect, further study is required to assess their possible risks in detail.

  12. Assessment of toxicity of radioactively contaminated sediments of the Yenisei River for aquatic plants in laboratory assay

    Energy Technology Data Exchange (ETDEWEB)

    Zotina, T.; Trofimova, E.; Medvedeva, M.; Bolsunovsky, A. [Institute of Biophysic SB RAS (Russian Federation)


    The Yenisei River has been subjected to radioactive contamination due to the operation of the Mining-and-Chemical Combine (Rosatom) (MCC) producing weapon-grade plutonium for more than fifty years (1958-2010). As a result, high activities of long-lived artificial radionuclides (Cs-137, Pu-238, 239, 241, Am-241) were deposited in sediments of the river. Bottom sediments of the Yenisei River downstream of the Krasnoyarsk city are also polluted with heavy metals because of industrial discharges and from the water catchment area. The purpose of this research was to estimate the ability of submersed macrophytes Elodea canadensis and Myriophyllum spicatum to serve as indicators of toxicity of bottom sediments of the Yenisei River. Activities of artificial radionuclides in the biomass of aquatic plants sampled in the Yenisei River upstream of the MCC were below detection limit (< 0.5 Bq/kg of dry mass for Cs-137). The activities of artificial radionuclides in the biomass of macrophytes sampled in the Yenisei River in the vicinity of the MCC in autumn 2012 were (Bq/kg of dry mass): 67±4 for Co-60, 16±2 for Cs-137, and 8±1 for Eu-152. For eco-toxicological experiments, top 20-cm layers of bottom sediments (BS) were collected from the Yenisei River at three sites in the vicinity of the MCC (No. 2-4) and at one site upstream of the MCC (No. 1). Samples of sediments contained natural isotope K-40 (240-330 Bq/kg, fresh mass) and artificial radionuclides: Co-60 (up to 70 Bq/kg), Cs-137 (0.8-1400 Bq/kg), Eu-152, 154 (up to 220 Bq/kg), Am-241 (up to 40 Bq/kg). The total activity concentration of radionuclides measured on an HPGe-Gamma-spectrometer (Canberra, U.S.) in samples of BS No. 1-4 was 330, 500, 880 and 1580 Bq/kg of fresh mass, respectively. Apical shoots of submersed macrophytes were planted in sediments (6-9 shoots per sediment sub-sample in three replicates). Endpoints of shoot and root growth were used as toxicity indicators; the number of cells with chromosome

  13. 洪泽湖湿地主要植物群落的水质净化能力研究%Water Quality Purification Ability of Main Wetland Plant Community in Hongze Lake

    Institute of Scientific and Technical Information of China (English)

    南楠; 张波; 李海东; 张金池


    通过建立围隔实验区的方法,以洪泽湖主要湿生植物群落芦苇、莲、菱、凤眼莲、苦草、金鱼藻为对象,研究了不同湿生植物群落对富营养化水体净化的能力.结果发现:研究区主要水生植物群落对富营养化水体总氮的去除能力从大到小依次为苦草(78.56%)、凤眼莲(66.26%)、金鱼藻(57.61%)、莲(50.18%),菱(46.04%)、芦苇(37.39%),对总磷的去除能力从大到小依次为苦草(61.84%)、芦苇(46.55%),菱(44.83%)、金鱼藻(45.00%),莲(39.66%)、凤眼莲(25.17%),对CODMn的去除能力从大到小依次为金鱼藻(61.33%)、苦草(55.56%),菱(52.44%),莲(47.11%)、凤眼莲(27.77%)、芦苇(23.33%).沉水植物苦草、金鱼藻对各种营养元素的净化效果都较好,产氧能力也较高;浮叶植物菱的净化效果比较稳定;挺水植物芦苇对总磷的净化效果稍好,而对其他各项净化能力均相对较弱.%The quality purification ability of different hygrophytes communities to eutrophication water was studied through the experimental enclosure of Hongze lake, and the different hygrophytes communities are Phragmites australis , Nelumbo nucifera , Trapa bispinosa , Eichhornia crassipes , Vallisneria natans and Ceratophyllum demersum. The results showed that the elimination ability of main wetland plant community to TN in eutrophication water body follwing the order of Vallisneria natans(78.56%), Eichhornia crassipes (66.26 % ), Ceratophyllum demersum (57.61%), Nelumbo nucifera (50.18%), Trapa bispinosa (46.04%),Phragmites australis (37. 39%), the elimination ability to TP follwing the order of Vallisneria natans (61.84%), Phragmites australis (46. 55%), Trapa bispinosa (44. 83%), Ceratophyllum demersum (45. 00%),Nelumbo nucifera (39.66%), Eichhornia crassipes (25.17%), the elimination ability to CODMn follwing the order of Ceratophyllum demersum (61.33%), Vallisneria natans (55. 56%), Trapa bispinosa (52.44%), Nelumbo nucifera

  14. Analysis and treatment of trouble of molecular sieve purification system of 21000 m^3/h air separation plant%21000m^3/h空分设备分子筛纯化系统故障分析及处理

    Institute of Scientific and Technical Information of China (English)

    胥波; 闫伟东; 孔繁魁; 沈荣; 田现德


    In the air separation plant with molecular sieve adsorption and purification process, the work condition of molecular sieve purification system restricts the safe and stable run of the entire air separation system. For the troubles frequently occurring in molecular sieve purification system, such as mechanical trouble of switching over valve, valve action-generated program problem, and program-interlocked control of molecular sieve purification system, the judgment of the trouble causes and treatment measures are detailed.%分子筛吸附净化流程空分设备中,分子筛纯化系统的运行工况制约了整个空分系统的安全稳定运行。针对分子筛纯化系统经常出现的切换阀机械故障、阀门动作导致的程序问题以及分子筛纯化系统程序联锁控制等问题,详细介绍了故障原因判断和处理措施。

  15. 四种室内观叶植物离体叶片吸收甲醛效果的研究%Formaldehyde Purification Effects of Four kinds of Indoor Foliage Plants Leaves in Vitro

    Institute of Scientific and Technical Information of China (English)

    张春晓; 刘梦云; 张莉莉; 熊勉


    Four common indoor ornamental plants were selected as experiment materials: Aglaonema modestum, Scindapsus aureun , Chlorophytum comosum , Hedera nepalensis var .sinensis.Through artificial simulation environment , explore formaldehyde purification effects of indoor ornamental plants in three different formaldehyde concentrations .The results showed that all the four plant species had a pretty good absorption capacity of formaldehyde .By comparing the absorption rate of formaldehyde with three different concentrations in 36 hours, Aglaonema modestum was the best , reached 23.94mg/cm2 , followed by Chlorophytum comosum and Hedera nepalensis var .sinensi, Scindapsus aureum was the weakest , formaldehyde absorption amount is 4.72mg/cm2 .Analyzing experimental data with statistics software spss , the results showed that each plant in vitro leave has different absorptive capacity after different time , and different plants have different absorptive capacity .Analyzing the process of adsorption formaldehyde by Chlorophytum , found that the adsorptive process is first-order.%以万年青、常春藤、吊兰、绿萝4种植物为研究对象,通过人工模拟环境,在3种不同的甲醛浓度下探讨4种植物离体吸收甲醛的能力。结果表明,植物离体叶片对甲醛均有较好的吸收能力;综合考虑36 h内4种植物离体叶片对甲醛的吸收量,其中以万年青的吸收效果最好,达到23.94 mg/cm2,吊兰和常春藤次之,绿萝效果最差,为4.72 mg/cm2。应用SPSS对实验数据进行分析,结果说明不同的处理时间下,各个植物离体叶片对甲醛的吸收能力都不同,且对甲醛的吸收能力因植物种类的不同而不同。对吊兰吸附甲醛的过程进行动力学分析,发现该吸附过程属于一级动力学过程。

  16. Utilization of flash gas in natural gas purification plant and its energy-saving measures%天然气净化厂闪蒸气利用节能措施探讨

    Institute of Scientific and Technical Information of China (English)

    郭戈; 张丽凤; 李春亮; 李岳峰; 黄东江; 黄海强; 陈武


    This paper mainly discusses the utilization of flash gas in natural gas purification plants so as to solve the liquid-carting problem of flash gas and avoid the waste of a great of combustible gases directly vented to flare in winter,and points out that this problem can be solved to assure the proper utilization of flash gas in winter through proper technical transformation.%天然气净化厂净化装置在冬季运行中,闪蒸气因带液影响燃料气系统运行而直接放空至火炬燃烧,浪费了大量可燃气体。本文主要讨论闪蒸气的利用问题,通过适当的工艺技术改造,解决天然气净化厂净化装置闪蒸气带液问题,确保闪蒸气在冬季正常利用。

  17. 浅议丙酮肟在引进分厂锅炉给水系统中的应用%Application of acetone oxime in the boiler feed system in Injin Natural Gas Purification Plant

    Institute of Scientific and Technical Information of China (English)

    唐浠; 瞿杨; 蒋宇; 吴高亮; 秦婧


    Because of the reasons that hydrazine is flammable and explosive ,slowly reacts with oxygen ,toxic ,etc .,it is substituted by acetone oxime as deoxidant in Yinjin Branch of Chongqing Natural Gas Purification Plant General .Analysis results show that the acetone oxime has better deoxidization effect and lower toxicity than hydrazine ,and it does not affect the water vapor system ,so that it can protect the equipment and has inhibit effect .Further more ,acetone oxime can also reduce the production cost ,therefore ,it is more economical and practical .%重庆天然气净化总厂引进分厂原来在锅炉给水系统中一直使用联氨作为除氧剂,但由于联氨易燃易爆、除氧速度慢、具有毒性等原因,则采用丙酮肟代替联氨进行除氧。分析使用丙酮肟除氧后的数据,丙酮肟较联氨而言,除氧效果更好,毒性更低,不影响水汽系统,对设备有保护作用和缓蚀效果,并减少了生产成本,更具有经济效益和实用性。

  18. 高硫煤制甲醇净化废气回收利用制取食品级液体CO2%Recovery and Utilization of Purification Waste Gas of High Sulfur Coal for Methanol Plant

    Institute of Scientific and Technical Information of China (English)

    赵绍武; 张萍; 汪俊


    The waste gas of purification unit was recovered and utilized in the high sulfur coal to methanol plant. Using domestic advanced and efficient technology, the carbon dioxide content in waste gas was recovered and purified to obtain food grade liquid carbon dioxide. The problems found in operation were timely analysed and optimized. The construction and operation of the device realized energy saving and emission reduction of CO2 in coal chemical industry and meet the green, low - carbon sustainable development train of thought.%高硫煤制甲醇净化废气回收利用装置,采用国内先进技术高效回收净化废气中二氧化碳,制取食品级液体CO2,对装置开车过程中发现的问题及时分析优化整改。该装置建设运行,实现了CO2节能减排,符合煤化工产业绿色、低碳可持续发展思路。

  19. A general method for assaying homo-and hetero-transglycanase activities that act on plant cell-wall polysaccharides

    Institute of Scientific and Technical Information of China (English)

    Lenka Frankova; Stephen C. Fry


    Transglycanases (endotransglycosylases) cleave a polysaccharide (donor-substrate) in mid-chain, and then transfer a portion onto another poly- or oligosaccharide (acceptor-substrate). Such enzymes contribute to plant cell-wall assembly and/or re-structuring. We sought a general method for revealing novel homo- and hetero-trans-glycanases, applicable to diverse polysaccharides and oligosaccharides, separating transglycanase-generated 3H-polysaccharides from unreacted 3H-oligosaccharides—the former immobilized (on filter-paper, silica-gel or glass-fiber), the latter eluted. On filter-paper, certain polysaccharides [e.g. (1!3, 1!4)-b-D-glucans] remained satisfactorily adsorbed when water-washed; others (e.g. pectins) were partially lost. Many oligosaccharides (e.g. arabinan-, galactan-, xylo-glucan-based) were successfully eluted in appropriate sol-vents, but others (e.g. [3H]xylohexaitol, [3H]mannohexaitol [3H]cellohexaitol) remained immobile. On silica-gel, all 3H-oligosaccharides left an immobile‘ghost’ spot (contaminating any 3H-polysaccharides), which was diminished but not prevented by additives e.g. sucrose or Triton X-100. The best stratum was glass-fiber (GF), onto which the reaction-mixture was dried then washed in 75%ethanol. Washing led to minimal loss or lateral migration of 3H-polysaccharides if conducted by slow percolation of acidified ethanol. The effectiveness of GF-blotting was well demonstrated for Chara vulgaris trans-b-mannanase. In conclusion, our novel GF-blotting technique efficiently frees transglycanase-gener-ated 3H-polysaccharides from unreacted 3H-oligosaccharides, enabling high-throughput screening of multiple postulated transglycanase activities utilising chemically diverse donor-and acceptor-substrates.

  20. Oestrogenic activity of a textile industrial wastewater treatment plant effluent evaluated by the E-screen test and MELN gene-reporter luciferase assay

    Energy Technology Data Exchange (ETDEWEB)

    Schiliro, Tiziana, E-mail: [Department of Public Health and Microbiology, University of Torino, Via Santena 5bis, 10126 Torino (Italy); Porfido, Arianna [Department of Public Health and Microbiology, University of Torino, Via Santena 5bis, 10126 Torino (Italy); Spina, Federica; Varese, Giovanna Cristina [Department of Life Sciences and Systems Biology, University of Torino, Viale Mattioli 25, 10125 Torino (Italy); Gilli, Giorgio [Department of Public Health and Microbiology, University of Torino, Via Santena 5bis, 10126 Torino (Italy)


    This study quantified the biological oestrogenic activity in the effluent of a textile industrial wastewater treatment plant (IWWTP) in northwestern Italy. Samples of the IWWTP effluent were collected monthly, both before and after tertiary treatment (ozonation). After solid phase extraction, all samples were subjected to two in vitro tests of total estrogenic activity, the human breast cancer cell line (MCF-7 BUS) proliferation assay, or E-screen test, and the luciferase-transfected human breast cancer cell line (MELN) gene-reporter assay, to measure the 17{beta}-oestradiol equivalent quantity (EEQ). In the E-screen test, the mean EEQ values were 2.35 {+-} 1.68 ng/L pre-ozonation and 0.72 {+-} 0.58 ng/L post-ozonation; in the MELN gene-reporter luciferase assay, the mean EEQ values were 4.18 {+-} 3.54 ng/L pre-ozonation and 2.53 {+-} 2.48 ng/L post-ozonation. These results suggest that the post-ozonation IWWTP effluent had a lower oestrogenic activity (simple paired t-tests, p < 0.05). The average reduction of estrogenic activity of IWWTP effluent after ozonation was 67 {+-} 26% and 52 {+-} 27% as measured by E-screen test and MELN gene-reporter luciferase assay, respectively. There was a positive and significant correlation between the two tests (Rho S = 0.650, p = 0.022). This study indicates that the environmental risk is low because oestrogenic substances are deposited into the river via IWWTP at concentrations lower than those at which chronic exposure has been reported to affect the endocrine system of living organisms. -- Highlights: Black-Right-Pointing-Pointer The two in vitro tests are suited for oestrogenic activity assessment in textile WWTP. Black-Right-Pointing-Pointer There is a significant correlation between the results of the two in vitro tests. Black-Right-Pointing-Pointer The oestrogenic activity of the effluent is reduced by ozonation. Black-Right-Pointing-Pointer The input of estrogenic substances into the river via textile WWTP is low.

  1. United purification effect of aquatic plants-electrolysis on azo dye wastewater%偶氮染料废水的水生植物-电解联合净化效应研究

    Institute of Scientific and Technical Information of China (English)

    丁绍兰; 杨冰; 朱超; 樊文娟


    结合电解处理技术和植物修复技术理论,建立了植物-电解联合净化系统,并用于对混合偶氮染料溶液的处理实验。结果表明,偶氮染料废水经植物-电解联合净化后水质更加稳定,脱色率和COD去除率最高分别可达98.7%和85%;出水微生物抑制率和生物抗过氧化能力显著降低,出水苯二胺和芳香胺类物质残留最大削减量分别为81.25%和99.17%。%Combining electrolysis treatment technique with phytoremediation technique ,the plants-electrolysis united purification system has been set up and used for the treatment experiments of mixed azo dye solution. The re-sults show that after azo dye wastewater has been purified jointly by plant-electrolysis process ,the water quality be-comes more steady. Its maximum decolorization rate and COD removing rate can reach 98.7%and 85%,respective-ly. The effluent microorganism inhibiting rate and biological over-oxidation ability are decreased obviously. The ma-ximum reduction rates of effluent diphenylamine and aromatic amines substance residues are 81.25%and 99.17%, respectively.

  2. TaqMan real-time PCR assays to assess arbuscular mycorrhizal responses to field manipulation of grassland biodiversity: effects of soil characteristics, plant species richness, and functional traits. (United States)

    König, Stephan; Wubet, Tesfaye; Dormann, Carsten F; Hempel, Stefan; Renker, Carsten; Buscot, François


    Large-scale (temporal and/or spatial) molecular investigations of the diversity and distribution of arbuscular mycorrhizal fungi (AMF) require considerable sampling efforts and high-throughput analysis. To facilitate such efforts, we have developed a TaqMan real-time PCR assay to detect and identify AMF in environmental samples. First, we screened the diversity in clone libraries, generated by nested PCR, of the nuclear ribosomal DNA internal transcribed spacer (ITS) of AMF in environmental samples. We then generated probes and forward primers based on the detected sequences, enabling AMF sequence type-specific detection in TaqMan multiplex real-time PCR assays. In comparisons to conventional clone library screening and Sanger sequencing, the TaqMan assay approach provided similar accuracy but higher sensitivity with cost and time savings. The TaqMan assays were applied to analyze the AMF community composition within plots of a large-scale plant biodiversity manipulation experiment, the Jena Experiment, primarily designed to investigate the interactive effects of plant biodiversity on element cycling and trophic interactions. The results show that environmental variables hierarchically shape AMF communities and that the sequence type spectrum is strongly affected by previous land use and disturbance, which appears to favor disturbance-tolerant members of the genus Glomus. The AMF species richness of disturbance-associated communities can be largely explained by richness of plant species and plant functional groups, while plant productivity and soil parameters appear to have only weak effects on the AMF community.

  3. 建筑室内植物对空气净化能力的研究——2010年上海世博会"沪上·生态家"植物筛选%Study on Indoor Air Purification Ability of Plants-Plants Screening for Shanghai Eco-home of the World Expo 2010 China Shanghai

    Institute of Scientific and Technical Information of China (English)



    2010年世博会以"城市,让生活更美好"为主题,"城市最佳实践区"的首次设立是本次世博会的创举和亮点之一,"沪上·生态家"是唯一代表上海参展2010年上海世博会的实物案例项目.为达到环境宜居和室内环境达标率100%的目标,同时满足(GB/T 50378-2006)三星级绿色建筑标准,"沪上·生态家"团队以目前市场上较为常用的室内观赏植物为测试对象,建立了测试方法以研究其叶片部分在模拟甲醛超标环境中的甲醛净化效果,为"沪上·生态家"的植物筛选提供试验依据.研究发现不同植物对甲醛的吸收能力不同,净化效果存在差异:在24 h内试验的50种植物中,迷迭香单位叶面积净化甲醛量最多,黄金葛绿萝净化率最大;同时植物会使植物测试舱内的湿负荷增加.在测试结果的基础上,综合景观效益,选取了部分植物作为上海世博会"沪上·生态家"的室内观赏植物.%The theme of the World Expo 2010 China Shanghai was better city, better life, and the establishment of urban best practices area (UBPA) was one of the original and bright spot works.In UBPA, Shanghai Eco-home was the only selected project represented Shanghai to join the Expo 2010.In order to satisfy the Evaluation Standard for Green Building (GB/T 50378-2006) and achieve the livable environment, the market commonly used indoor ornamental plants were taken as the research objects, the purification effects of plants' leaves in the simulated excessive formaldehyde environment were studied.The measured results indicated that different plants had different absorption capacities of formaldehyde, also for the purification abilities.During the test period of 24 h and for all the 50 kinds of plants,the unit leaf area of rosemary had the largest ability to purify the formaldehyde and the scindapsus aureum had the largest purification rate.Meanwhile, the moisture load in the test chamber was increased due to the plants.Based on

  4. Heterologous Expression of Three Plant Serpins with Distinct Inhibitory Specificities

    DEFF Research Database (Denmark)

    Dahl, Søren Weis; Rasmussen, Søren Kjærsgård; Hejgaard, Jørn


    For the first time, inhibitory plant serpins, including WSZ1 from wheat, BSZ4, and the previously unknown protein BSZx from barley, have been expressed in Escherichia coli, and a procedure for fast purification of native plant serpins has been developed, BSZx, BSZ4, and WSZ1 were assayed for inhi...... favorable P-2 Leu. BSZ4 inhibited cathepsin G (k(a) = 2.7 x 10(4) M(-1) s(-1)) at P-1 Met but was hydrolyzed by trypsin and chymotrypsin. The three plant serpins formed stable SDS-resistant complexes with the proteinases in accordance with the kinetic data.......For the first time, inhibitory plant serpins, including WSZ1 from wheat, BSZ4, and the previously unknown protein BSZx from barley, have been expressed in Escherichia coli, and a procedure for fast purification of native plant serpins has been developed, BSZx, BSZ4, and WSZ1 were assayed......, the apparent rate constant for chymotrypsin inhibition at P-2 (k(a) = 9.4 x 10(5) M(-1) s(-1)) was only four times lower than for trypsin at P-1 (k(a) = 3.9 X 10(6) M(-1) s(-1)), and the apparent inhibition stoichiometries were close to 1. Furthermore, our data suggest that cathepsin G was inhibited by BSZx (k...

  5. Development of Purification Protocol Specific for Bacteriocin 105B (United States)


    2007. 45-92. Levy, S.B. “Factors Impacting on the Problem of Antibiotic Resistance ”. J. Antimicrob. Chermother. 49(1): 25-30 (2002). Levy...Bacillus anthracis. As the current application of broad-spectrum antimicrobials promotes the development of multi- drug resistant microorganisms...SPECTRUM TARGETED ANTIMICROBIALS ASSAYS PURIFICATION BACILLUS ANTHRACIS DRUG- RESISTANT MICROORGANISMS

  6. Impact of ubiquitous inhibitors on the GUS gene reporter system: evidence from the model plants Arabidopsis, tobacco and rice and correction methods for quantitative assays of transgenic and endogenous GUS

    Directory of Open Access Journals (Sweden)

    Gerola Paolo D


    Full Text Available Abstract Background The β-glucuronidase (GUS gene reporter system is one of the most effective and employed techniques in the study of gene regulation in plant molecular biology. Improving protocols for GUS assays have rendered the original method described by Jefferson amenable to various requirements and conditions, but the serious limitation caused by inhibitors of the enzyme activity in plant tissues has thus far been underestimated. Results We report that inhibitors of GUS activity are ubiquitous in organ tissues of Arabidopsis, tobacco and rice, and significantly bias quantitative assessment of GUS activity in plant transformation experiments. Combined with previous literature reports on non-model species, our findings suggest that inhibitors may be common components of plant cells, with variable affinity towards the E. coli enzyme. The reduced inhibitory capacity towards the plant endogenous GUS discredits the hypothesis of a regulatory role of these compounds in plant cells, and their effect on the bacterial enzyme is better interpreted as a side effect due to their interaction with GUS during the assay. This is likely to have a bearing also on histochemical analyses, leading to inaccurate evaluations of GUS expression. Conclusions In order to achieve reliable results, inhibitor activity should be routinely tested during quantitative GUS assays. Two separate methods to correct the measured activity of the transgenic and endogenous GUS are presented.

  7. 从分子筛纯化系统的设计及操作谈空分设备节能%Approach to energy-saving for air separation plant from the design and operation of molecular sieve purification system

    Institute of Scientific and Technical Information of China (English)



    The energy-saving for molecular sieve purification system of air separation plant is analyzed from the process design of molecular sieve purification system, the design and selection of equipment, the engineering design and its operation and maintenance, and%从分子筛纯化系统的流程设计、设备的设计与选型、工程设计及其操作、维护等方面,分析空分设备分子筛纯化系统的节能问题,提出了对节能措施的一些思考和建议。


    Directory of Open Access Journals (Sweden)

    L. Pilinevich


    Full Text Available The paper presents results of the investigations on the water purification process with the help of photocatalysis using the photocatalyst which is developed on the basis of porous titanium with the layer of nanoparticle layer of titanium dioxide and an experimental plant. The investigations results have shown high efficiency of the developed photocatalytic materials and a water purification plants

  9. Expression and Purification of ST14, a Tumor Metastasis-associated Protein, and Its Activity Assay%肿瘤转移相关蛋白ST14的表达、纯化及活性鉴定

    Institute of Scientific and Technical Information of China (English)

    葛维挺; 郑树; 孙立峰; 史影; 胡涵光; 丁克峰


    ST14 is one of the type Ⅱ transmembrane serine proteases that correlates with the process of tumor metastasis. The C-terminal catalytic region (900 bp) of ST14 was cloned into the expression vector pGEX-4T-2 and the positive plasmid pGEX-4T-2-STI4 was transformed into E. coli BL21, then cultured and induced with IPTG. The chaperonin GroEL was found to be tightly associated with the fusion protein and co-purified with it by regular GST affinity chromatography. A method for the removal of contaminating GroEL from GST-ST14 fusion protein was described, the purity of product was 96.2%. Enzyme activity assay indicated that this fusion protein had serine protease activity.

  10. Purification of equine Gc-globulin

    DEFF Research Database (Denmark)

    Houen, Gunnar; Pihl, Tina Holberg; Andersen, Pia Haubro

    Objectives With the aim of producing antibodies for an equine Group specific component (Gc)-globulin assay, the protein was purified from normal equine plasma. Methods Equine Gc-globulin was purified from healthy horse plasma using ion exchange chromatography (Q-Sepharose, CM......-Sepharose) and preparative PAGE. Results Equine Gc-globulin has successfully been purified from healthy horse plasma and rabbits and mice are being immunized to produce specific antibodies. Conclusions Purification of equine Gc-globulin and the production of specific antibodies will make it possible to develop an assay...... to be a sensitive marker of acute tissue injury and fatal outcome in humans. Patients with a low plasma concentration of Gc-globulin due to severe tissue injury might potentially benefit from infusions with purified Gc-globulin [1]. With an equine Gc-globulin assay, future studies will investigate the concentration...

  11. New Gateway-compatible vectors for a high-throughput protein-protein interaction analysis by a bimolecular fluorescence complementation (BiFC) assay in plants and their application to a plant clathrin structure analysis. (United States)

    Nishimura, Kohji; Ishikawa, Syouta; Matsunami, Erika; Yamauchi, Junji; Homma, Keiichi; Faulkner, Christine; Oparka, Karl; Jisaka, Mitsuo; Nagaya, Tsutomu; Yokota, Kazushige; Nakagawa, Tsuyoshi


    Protein-protein interactions (PPI) play key roles in various biological processes. The bimolecular fluorescence complementation (BiFC) assay is an excellent tool for routine PPI analyses in living cells. We developed new Gateway vectors for a high-throughput BiFC analysis of plants, adopting a monomeric Venus split just after the tenth β-strand, and analyzed the interaction between Arabidopsis thaliana coated vesicle coatmers, the clathrin heavy chain (CHC), and the clathrin light chain (CLC). In competitive BiFC tests, CLC interacted with CHC through a coiled-coil motif in the middle section of CLC. R1340, R1448, and K1512 in CHC and W94 in CLC are potentially key amino acids underlying the inter-chain interaction, consistent with analyses based on homology modeling. Our Gateway BiFC system, the V10-BiFC system, provides a useful tool for a PPI analysis in living plant cells. The CLC-CHC interaction identified may facilitate clathrin triskelion assembly needed for cage formation.

  12. Production, purification, and capsid stability of rhinovirus C types. (United States)

    Griggs, Theodor F; Bochkov, Yury A; Nakagome, Kazuyuki; Palmenberg, Ann C; Gern, James E


    The rhinovirus C (RV-C) were discovered in 2006 and these agents are an important cause of respiratory morbidity. Little is known about their biology. RV-C15 (C15) can be produced by transfection of recombinant viral RNA into cells and subsequent purification over a 30% sucrose cushion, even though yields and infectivity of other RV-C genotypes with this protocol are low. The goal of this study was to determine whether poor RV-C yields were due to capsid instability, and moreover, to develop a robust protocol suitable for the purification of many RV-C types. Capsid stability assays indicated that virions of RV-C41 (refractory to purification) have similar tolerance for osmotic and temperature stress as RV-A16 (purified readily), although C41 is more sensitive to low pH. Modification to the purification protocol by removing detergent increased the yield of RV-C. Addition of nonfat dry milk to the sucrose cushion increased the virus yield but sacrificed purity of the viral suspension. Analysis of virus distribution following centrifugation indicated that the majority of detectable viral RNA (vRNA) was found in pellets refractory to resuspension. Reduction of the centrifugal force with commiserate increase in spin-time improved the recovery of RV-C for both C41 and C2. Transfection of primary lung fibroblasts (WisL cells) followed by the modified purification protocol further improved yields of infectious C41 and C2. Described herein is a higher yield purification protocol suitable for RV-C types refractory to the standard purification procedure. The findings suggest that aggregation-adhesion problems rather than capsid instability influence RV-C yield during purification.

  13. Purification and Structural and Kinetic Characterization of the Pyrophosphate:Fructose-6-Phosphate 1-Phosphotransferase from the Crassulacean Acid Metabolism Plant, Pineapple. (United States)

    Tripodi, KEJ.; Podesta, F. E.


    Pyrphosphate-dependent phosphofructokinase (PFP) was purified to electrophoretic homogeneity from illuminated pineapple (Ananas comosus) leaves. The purified enzyme consists of a single subunit of 61.5 kD that is immunologically related to the potato tuber PFP [beta] subunit. The native form of PFP likely consists of a homodimer of 97.2 kD, as determined by gel filtration. PFP's glycolytic activity was strongly dependent on pH, displaying a maximum at pH 7.7 to 7.9. Gluconeogenic activity was relatively constant between pH 6.7 and 8.7. Activation by Fru-2,6-bisphosphate (Fru-2,6-P2) was dependent on assay pH. In the glycolytic direction, it activated about 10-fold at pH 6.7, but only 2-fold at pH 7.7. The gluconeogenic reaction was only weakly affected by Fru-2,6-P2. The true substrates for the PFP forward and reverse reactions were Fru-6-phosphate and Mg-pyrophosphate, and Fru-1,6-P2, orthophosphate, and Mg2+, respectively. The results suggest that pineapple PFP displays regulatory properties consistent with a pH-based regulation of its glycolytic activity, in which a decrease in cytosolic pH caused by nocturnal acidification during Crassulacean acid metabolism, which could curtail its activity, is compensated by a parallel increase in its sensitivity to Fru-2,6-P2. It is also evident that the [beta] subunit alone is sufficient to confer PFP with a high catalytic rate and the regulatory properties associated with activation by Fru-2,6-P2.

  14. Studies on the dispersion behaviour of pneumatically metered additives in combustion chambers and flue gas purification plants; Untersuchungen zum Ausbreitungsverhalten pneumatisch zudosierter Additive in Brennkammern und Rauchgasreinigungsanlagen

    Energy Technology Data Exchange (ETDEWEB)

    Schaberg, U. [Stuttgart Univ. (Germany). Inst. fuer Mechanische Verfahrenstechnik


    In direct desulphurisation of power plants the pneumatically injected additive propagates into the flow of the reaction chamber as a dust-charged free jet of a mixture of air and solid particles. As the dispersion of the dust-charged free jet in the transverse flow has hardly been studied to data, there are no established criteria for the design of such installations. In this study the influences of the most important parameters on the dispersion behaviour of the charged free jets were examined. In particular, the distribution of solid matter within the cross-section of the jet and its change along the path of propagation were measured. Glass dust, silicium carbide, and quartz powder were used as test dusts. Comemrcial additives are not suitable for experimental work as their small grain size makes them difficult to handle. (orig.) [Deutsch] Bei Additiveinblasesystemen fuer die Direktentschwefelung in Kraftwerken breitet sich das Feststoff/Luft-Gemisch als staubbeladener Freistrahl in der Grundstroemung des Reaktionsraumes aus. Die Ausbreitung der staubbeladenen, querangestroemten Freistrahlen wurde bisher praktisch nicht untersucht, so dass gesicherte Auslegungskriterien fuer technische Anwendungen fehlen. In der vorliegenden Arbeit wurde die Wirkung der wichtigsten Parameter auf das Ausbreitungsverhalten der beladenen Freistrahlen untersucht. Vor allem wurde die Feststoffverteilung im Strahlquerschnitt und ihre Entwicklung laengs dem Strahlweg gemessen. Dazu wurden Glasstaub, Siliziumkarbid und Quarzmehl als Versuchsstaeube verwendet. Der Einsatz technisch gebraeuchlicher Additive scheitert an ihrer feinen Koernung und der damit verbundenen schlechten Handhabung im Versuchsbetrieb. (orig.)

  15. Purification and characterization of alkaline pectin lyase from a newly isolated Bacillus clausii and its application in elicitation of plant disease resistance. (United States)

    Li, Zuming; Bai, Zhihui; Zhang, Baoguo; Li, Baojv; Jin, Bo; Zhang, Michael; Lin, Francis; Zhang, Hongxun


    Alkaline pectin lyase (PNL) shows potential as a biological control agent against several plant diseases. We isolated and characterized a new Bacillus clausii strain that can produce 4,180 U/g of PNL using sugar beet pulp as a carbon source and inducer. The PNL was purified to apparent homogeneity using ultrafiltration, ammonium sulfate fractionation, DEAE Sepharose Fast Flow, and Sephadex G-75 gel filtration. The purified PNL was found to be a monomeric protein with a molecular weight of 35 kDa, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). It demonstrated optimal activity with K(m) of 0.87 mg/ml at pH 10.0 and 60 °C. The enzyme is stable in the pH range of 8.0-10.0 and temperature ≤40 °C. Ca(2+) was found to stimulate the enzymatic activity of the PNL by up to 410 %. Mass spectrometric results gave 38 % match coverage with pectate lyase from B. clausii KSM-K16 (gi|56961845). The PNL was found to elicit disease resistance in cucumber seedlings, suggesting that it may have applications in biocontrol and sustainable agriculture.

  16. 环氧乙烷精制单元生产优化%The operation problems and optimization measures for purification unit of EO plant

    Institute of Scientific and Technical Information of China (English)

    张宏庆; 王鸿生; 梁宏伟; 李晓光


    介绍了吉林石化乙二醇厂乙二醇装置增设环氧乙烷精制单元项目后的优化运行情况,包括项目运行中的工艺运行优化、对运行存在问题的管线改造和停车清洗置换方案优化情况.通过对该单元的持续运行优化,极大提高了单元运行的综合效益.%This article introduces the situation on glycol plant joint production ethylene oxide(EO) project implementation unit,including operation and optimization of process operation,pipeline reform to solve the problem,the clean and replace program optimization.Late through the project implementation and operation optimization,we ensure that the unit running smoothly,greatly improve the comprehensive benefits of the unit running.

  17. 基于去污效果和气候适应性的湿地植物筛选研究%Wetland plant screening based on the purification effect and climate adaptability

    Institute of Scientific and Technical Information of China (English)

    苏小红; 汤晓玉; 顾新娇; 王文国; 何明雄; 胡启春


    通过模拟垂直潜流人工湿地系统,在对菖蒲(Acorus calamus)、芦竹(Arundo donax)、茭白(Zizania lati folia)、旱伞草(Cyperus alterni folius)和再力花(Thalia dealbata)5种湿地植物的去污能力进行分析的基础上,利用最大熵(MAXENT)模型对它们在中国西南地区的适生区和分布的气候影响因子进行了综合分析.结果表明,5种湿地植物都有较高的去污能力,且不同植物的去污能力存在一定差异,对氮、磷去除效果较好的为旱伞草,茭白、再力花和菖蒲次之,芦竹较差;旱伞草在中国西南地区有着广泛的适生区,并可以越冬,是一种去污能力较好并可以全年使用的人工湿地植物.菖蒲、茭白、再力花也有较强的去污能力和气候适应能力,但是过冬效果相对较差,在西南地区大部分区域的春、夏、秋季适用于人工湿地栽种.芦竹虽然在西南地区适生区较广,但它在淹水环境中的去污能力相对较弱,可以适当地应用于人工湿地周边的湿生环境中.%Through the simulation of vertical subsurface flow constructed wetland system,the decontamination capability of Acorus calamus,Arundo donax,Zizania lati folia,Cyperus alterni folius and Thalia dealbata were studied.MAXENT model was employed to analyze potential distribution area of the 5 wetland plants in Southwestern China and the main climate impact factors that influencing their distribution.The results showed that all the 5 wetland plants presented perfect water purification efficiency,however their contaminant removal capacity had some differences.Cyperus alternifolius,which has wide range of potential distribution in Southwestern China,had the best removal capacity and was able to live through the winter,thereby it was regarded as a preferable wetland plant that can be used through the whole year.Acorus calamus,Zizania latifolia and Thalia dealbata also had good removal capacity and climate adaptability,but they had poor

  18. Purification of GST-Tagged Proteins. (United States)

    Schäfer, Frank; Seip, Nicole; Maertens, Barbara; Block, Helena; Kubicek, Jan


    This protocol describes the purification of recombinant proteins fused to glutathione S-transferase (GST, GST-tagged proteins) by Glutathione Affinity purification. The GST tag frequently increases the solubility of the fused protein of interest and thus enables its purification and subsequent functional characterization. The GST-tagged protein specifically binds to glutathione immobilized to a matrix (e.g., agarose) and can be easily separated from a cell lysate by a bind-wash-elute procedure. GST-tagged proteins are often used to study protein-protein interactions, again making use of glutathione affinity in a procedure called a GST pull-down assay. The protocol is designed to process 200 ml of E. coli culture expressing intermediate to high amounts of a GST-tagged protein (~25 mg l(-1)). Depending on the expression rate or the available culture volume, the scale can be increased or decreased linearly. The protocol can also be used to purify GST-tagged proteins from other expression systems, such as insect or mammalian cells. Tips are provided to aid in modifying certain steps if proteins shall be recovered from alternative expression systems.

  19. Expression and purification of splicing proteins from mammalian cells. (United States)

    Allemand, Eric; Hastings, Michelle L


    Pre-mRNA splicing is a complex process that is carried out by a large ribonucleoprotein enzyme, termed the spliceosome, which comprises up to 200 proteins. Despite this complexity, the role of individual spliceosomal proteins in the splicing reaction has been successfully investigated using cell-free assays. In many cases, the splicing factor of interest must be expressed and purified in order to study its function in vitro. Posttranslational modifications such as phosphorylation, methylation, acetylation, and ubiquitination of splicing factors are important for activity. Thus, their purification from mammalian cells presents numerous advantages. Here, we describe a method for expression and purification of splicing proteins from mammalian cells.

  20. Purification of Murine Monoclonal IgM Antibody

    Institute of Scientific and Technical Information of China (English)


    This paper presents the purification of a monoclonal IgM antibody against human tumor associated antigen Lewis-Y by ion exchange chromatography and gel filtration.Enzyme-linked immunosorbent assay (ELISA) and SDS-polyacrylamide gel electrophoresis (PAGE) were used to identify purified IgM antibody.In flow cytometry analysis, the purified IgM antibody recognizes human breast tumor cell line MCF-7 which expresses Lewis-Y antigen.This work presents a new way for the purification of murine monoclonal IgM antibody.

  1. 植物净化空气中汞污染的研究%Study on Purification of Air Mercury Pollution by Plants

    Institute of Scientific and Technical Information of China (English)

    蒋蓉芳; 周德灏; 戴修道


    对上海市常见易生长的22种植物进行了现场采样测试,结果发现,瓜子黄杨、广玉兰、海桐、蚊母、墨西哥落叶杉及棕榈对汞蒸气具有很强的吸收富集能力及抗毒性能.然后采用富集能力最强的瓜子黄杨进行一系列试验,结果显示,汞浓度随接触瓜子黄杨时间的延长而降低,而瓜子黄杨叶片的汞含量随时间的延长而增加。根据吸汞实验,计算得瓜子黄杨叶面积的吸气速率为0.133m3/h·m2。%Through investigation on 22 specie of common plants in Shanghai, it was found that Littleleaf Box,Southern Magnolia, Tobira Pittosporum, Pacemose Distylium, Mexican Cypress and Fortune Windmillpalm have great ability to absorb and enrich mercury vapour and anti-toxicity. The following experiment on Littleleaf Box which has the greatest ability of enrichment showed that concentration of mercury in the leaves of Littleleaf Box increased while the concentration of mercury vapour in indoor air decreased, and according to the absorbing experiment, the absorbing rate reached 0.133m3/h·m2.

  2. Microcoupon Assay Of Adhesion And Growth Of Bacterial Films (United States)

    Pierson, Duane L.; Koenig, David W.


    Microbiological assay technique facilitates determination of some characteristics of sessile bacteria like those that attach to and coat interior walls of water-purification systems. Biofilms cause sickness and interfere with purification process. Technique enables direct measurement of rate of attachment of bacterial cells, their metabolism, and effects of chemicals on them. Used to quantify effects of both bactericides and growth-stimulating agents and in place of older standard plate-count and tube-dilution techniques.

  3. DNA staining with the fluorochromes EtBr, DAPI and YOYO-1 in the comet assay with tobacco plants after treatment with ethyl methanesulphonate, hyperthermia and DNase-I. (United States)

    Gichner, Tomás; Mukherjee, Anita; Velemínský, Jirí


    We applied the alkaline version of the single-cell gel electrophoresis (comet) assay to roots and leaves of tobacco (Nicotiana tabacum var. xanthi) seedlings or isolated leaf nuclei treated with: (1) the alkylating agent ethyl methanesulphonate, (2) necrotic heat treatments at 50 degrees C, and (3) DNase-I. All three treatments induced a dose-dependent increase in DNA migration, expressed as percentage of tail DNA. A comparison of the fluorochrome DNA dyes ethidium bromide, DAPI and YOYO-1 demonstrated that for the alkaline version of the comet assay in plants, the commonly used fluorescent dye ethidium bromide can be used with the same efficiency as DAPI or YOYO-1.

  4. Electron beam silicon purification

    Energy Technology Data Exchange (ETDEWEB)

    Kravtsov, Anatoly [SIA ' ' KEPP EU' ' , Riga (Latvia); Kravtsov, Alexey [' ' KEPP-service' ' Ltd., Moscow (Russian Federation)


    Purification of heavily doped electronic grade silicon by evaporation of N-type impurities with electron beam heating was investigated in process with a batch weight up to 50 kilos. Effective temperature of the melt, an indicative parameter suitable for purification process characterization was calculated and appeared to be stable for different load weight processes. Purified material was successfully approbated in standard CZ processes of three different companies. Each company used its standard process and obtained CZ monocrystals applicable for photovoltaic application. These facts enable process to be successfully scaled up to commercial volumes (150-300 kg) and yield solar grade silicon. (copyright 2014 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  5. Enzyme assays. (United States)

    Reymond, Jean-Louis; Fluxà, Viviana S; Maillard, Noélie


    Enzyme assays are analytical tools to visualize enzyme activities. In recent years a large variety of enzyme assays have been developed to assist the discovery and optimization of industrial enzymes, in particular for "white biotechnology" where selective enzymes are used with great success for economically viable, mild and environmentally benign production processes. The present article highlights the aspects of fluorogenic and chromogenic substrates, sensors, and enzyme fingerprinting, which are our particular areas of interest.

  6. Effects of two plant growth regulators, indole-3-acetic acid and β-naphthoxyacetic acid, on genotoxicity in Drosophila SMART assay and on proliferation and viability of HEK293 cells from the perspective of carcinogenesis. (United States)

    Karadeniz, Asuman; Kaya, Bülent; Savaş, Burhan; Topcuoğlu, Ş Fatih


    In this study, the mutagenic and recombinogenic effects of indole-3-acetic acid (IAA), a plant growth regulator naturally synthesized in plants but produced synthetically, and β-naphthoxyacetic acid (BNOA), a synthetic plant growth regulator widely used in agricultural regions, were investigated using the somatic mutation and recombination test (SMART) in Drosophila wings. The effect of the same plant growth regulators against the proliferation and viability of a human immortalized embryonic kidney HEK293 cells which is at the early stage of carcinogenesis were also examined with MTT and trypan-blue exclusion assays. For the SMART assay, two different crosses were used: a standard and a high-bioactivation (HB) cross, involving the flare-3 and the multiple wing hairs markers. The HB cross involved flies characterized by an increased cytochrome P-450-dependent bioactivation capacity, which permits the more efficient biotransformation of promutagens and procarcinogens. In both crosses, the wings of the two types of progeny, inversion-free marker heterozygotes and balancer heterozygotes, were analyzed. The results show that IAA and BNOA are not mutagenic or recombinogenic in the wing cells of Drosophila. Furthermore, neither plant growth regulator affected the proliferation rate of HEK293 cells; however, both of them induced cell death at high concentrations.

  7. Quantitative and specific detection of the biocontrol agent, Serratia plymuthica, in plant extracts using a real-time TaqMan® assay

    NARCIS (Netherlands)

    Czajkowski, R.L.; Wolf, van der J.M.


    A Serratia plymuthica-specific TaqMan® assay was designed based on the consensus nucleotide sequence from the 3'- end of the luxS gene present in all S. plymuthica strains tested. The specificity of the assay was demonstrated by testing 21 Serratia spp. strains and 30 isolates belonging to various s

  8. 油田燃煤电厂CO2捕集纯化工程实践%CO2 capture and purification in coal-fired power plant

    Institute of Scientific and Technical Information of China (English)

    马玉峰; 郝杰; 万志鹏; 张翠红


    CO2是一种温室气体,通过CO2收集、驱油技术,能将造成温室效应的气体用于提高原油采收率,同时减少工业生产中温室气体的排放。为了实现CO2的捕集纯化,胜利油田采用一乙醇胺溶液( MEA)化学吸收工艺捕集CO2。介绍了“低渗透油藏CO2驱油”重大先导试验,在胜利发电厂建设CO2捕集纯化装置,通过此装置收集稳定、廉价的CO2气体用于驱油生产实践。通过分析系统运行状况,对装置进行了一系列的试验、研究,总结了大量CO2捕集系统的工程应用经验。胜利油田CO2捕集项目,通过将大型燃煤电厂烟道气中CO2捕集纯化、安全输送等系列技术攻关,形成低能耗捕集纯化、运输的集成配套技术。%CO2,known as a greenhouse gas,is recycled to improve the production of crude oil through collection an d oil-driving technolo-gies.Shengli Oilfield adopts MEA chemical adsorption process to capture and purify CO2.To develop low permeability reservoir,conduct CO2 flooding experiment.First,build capture and purification devices which capture and purify the CO2 from the large-scale coal-fired power plant.Analyze the matched CO2 conveying techniques.The actual operation experiment guides the subsequent object design.

  9. 特殊植物类群空气凤梨对大气污染物甲醛的净化%Purification of air pollutant-formaldehyde with special plant group-epiphytic Tillandsia

    Institute of Scientific and Technical Information of China (English)

    李俊霖; 李鹏; 王恒蓉; 郑桂灵


    Epiphytic Tillandsia species uptake nutrients and moistures directly from the atmosphere, so they are often used for monitoring atmospheric heavy metal pollutants and organic pollutants, but have not been applied to the study of purifying formaldehyde yet. Two species of Tillandsia, i. e. T. usneoides and T. stricta, as well as Chlorophytum malayense, were exploited to test their ability of removing formaldehyde under two different conditions, i. e. sealed glass boxes and closed laboratory. Although the morphology and physiology of these plants had some changes, no obvius damages were suffered under formaldehyde stress. After 6-8 hours of purification with three plants, two species of Tillandsia achieved similar results with Chlorophytum. However, the speed that Tillandsia purifying formaldehyde was much higher than Chlorophytum within first 2 hours, which may due to the existence of the foliar hydrophilic trichomes on the leave surface of Tillandsia. The above results suggested that Tillandsia could purify formaldehyde more rapidly and efficiently than Chlorophytum, and it could be applied to remove indoor formaldehyde pollutant.%空气凤梨是一类生长在空气中、不需要土壤、生长所需的水分和营养可以全部来自空气的特殊植物.它们常被用来指示与修复大气重金属污染物和有机污染物,但尚未应用于甲醛净化研究.为了探讨空气凤梨对甲醛的净化效果,我们以2种空气凤梨为实验材料,吊兰为对照材料,通过密封箱内甲醛熏蒸及在封闭的实际环境中进行了实验.结果表明,松萝铁兰、硬叶空凤和吊兰3种植物在甲醛胁迫下,外部形态和生理指标有一定的变化,但未受到明显的伤害.更重要的是,3种植物对甲醛均有相当强的净化作用.6~8h后,2种空气凤梨对甲醛的净化可达到与吊兰相近的效果.而在2h内,空气凤梨净化甲醛的速度远远大于吊兰,这可能是与空气凤梨叶片表面覆盖有亲水性

  10. Water Purification Product (United States)


    Ecomaster, an affiliate of BioServe Space Technologies, this PentaPure technology has been used to purify water for our nation's Space Shuttle missions since 1981. WTC-Ecomaster of Mirneapolis, Minnesota manufactures water purification systems under the brand name PentaPure (TM). BioServe researcher Dr. George Marchin, of Kansas State University, first demonstrated the superiority of this technology and licensed it to WTC. Marchin continues to perform microgravity research in the development of new technologies for the benefit of life on Earth.

  11. Effect of charcoal on water purification


    Suzuki, Hirotaka; Kawahigashi, Tatsuo


    [Abstract] A natural basin system purifies water through self-purification, but the water pollution load of a river might exceed its self-purification capacity. Charcoal, which is used for other uses aside from heating, such as air purification, was evaluated experimentally for water quality purification. The experiment described herein is based on simple water quality measurements. Some experimentally obtained results are discussed.

  12. Rover waste assay system

    Energy Technology Data Exchange (ETDEWEB)

    Akers, D.W.; Stoots, C.M.; Kraft, N.C.; Marts, D.J. [Idaho National Engineering Lab., Idaho Falls, ID (United States)


    The Rover Waste Assay System (RWAS) is a nondestructive assay system designed for the rapid assay of highly-enriched {sup 235}U contaminated piping, tank sections, and debris from the Rover nuclear rocket fuel processing facility at the Idaho Chemical Processing Plant. A scanning system translates a NaI(Tl) detector/collimator system over the structural components where both relative and calibrated measurements for {sup 137}Cs are made. Uranium-235 concentrations are in operation and is sufficiently automated that most functions are performed by the computer system. These functions include system calibration, problem identification, collimator control, data analysis, and reporting. Calibration of the system was done through a combination of measurements on calibration standards and benchmarked modeling. A description of the system is presented along with the methods and uncertainties associated with the calibration and analysis of the system for components from the Rover facility. 4 refs., 2 figs., 4 tabs.

  13. Purification and biochemical characterization of recombinant Persicaria minor β-sesquiphellandrene synthase (United States)

    Ker, De-Sheng; Pang, Sze Lei; Othman, Noor Farhan; Kumaran, Sekar; Tan, Ee Fun; Krishnan, Thiba; Chan, Kok Gan; Othman, Roohaida


    Background Sesquiterpenes are 15-carbon terpenes synthesized by sesquiterpene synthases using farnesyl diphosphate (FPP) as a substrate. Recently, a sesquiterpene synthase gene that encodes a 65 kDa protein was isolated from the aromatic plant Persicaria minor. Here, we report the expression, purification and characterization of recombinant P. minor sesquiterpene synthase protein (PmSTS). Insights into the catalytic active site were further provided by structural analysis guided by multiple sequence alignment. Methods The enzyme was purified in two steps using affinity and size exclusion chromatography. Enzyme assays were performed using the malachite green assay and enzymatic product was identified using gas chromatography-mass spectrometry (GC-MS) analysis. Sequence analysis of PmSTS was performed using multiple sequence alignment (MSA) against plant sesquiterpene synthase sequences. The homology model of PmSTS was generated using I-TASSER server. Results Our findings suggest that the recombinant PmSTS is mainly expressed as inclusion bodies and soluble aggregate in the E. coli protein expression system. However, the addition of 15% (v/v) glycerol to the protein purification buffer and the removal of N-terminal 24 amino acids of PmSTS helped to produce homogenous recombinant protein. Enzyme assay showed that recombinant PmSTS is active and specific to the C15 substrate FPP. The optimal temperature and pH for the recombinant PmSTS are 30 °C and pH 8.0, respectively. The GC-MS analysis further showed that PmSTS produces β-sesquiphellandrene as a major product and β-farnesene as a minor product. MSA analysis revealed that PmSTS adopts a modified conserved metal binding motif (NSE/DTE motif). Structural analysis suggests that PmSTS may binds to its substrate similarly to other plant sesquiterpene synthases. Discussion The study has revealed that homogenous PmSTS protein can be obtained with the addition of glycerol in the protein buffer. The N-terminal truncation

  14. Control of higher alkynes in purification process of acetylene in natural gas to acetylene plant%浅谈天然气制乙炔净化装置中高级炔含量的控制

    Institute of Scientific and Technical Information of China (English)



    介绍了乙炔净化工艺和乙炔中甲基乙炔、1,3-丁二烯、丙二烯等高级炔脱除原理,分析了影响它们脱除的因素,在此基础上提出了控制乙炔中高级炔含量的方法并实施,实现了乙炔净化装置高负荷、长周期、稳定运行。%The purification process of acetylene and the principle of removing methyl acetylene, 1,3-butadiene, allene and other higher alkynes from acetylene were introduced, and the factors influencing the purification process were analyzed. Based on the analysis, the measures for controlling the level of higher alkynes in acetylene were proposed and implemented, which ensured that the acetylene purification unit could run in high load, long period and stable operation.

  15. The controllability analysis of the purification system for heavy water reactors

    Energy Technology Data Exchange (ETDEWEB)

    Lee, K. D.; Cho, B. H.; Shin, C. H.; Kim, S. H. [KEPRI, Taejon (Korea, Republic of); Lee, Y. K.; Kim, K. U. [KHNP, Kyungju (Korea, Republic of)


    The heavy water reactor such as Wolsung No.1 and No.2 has a purification system to purify the reactor coolant. The control system regulates the coolant temperature to protect the ion exchanger. After the fuel exchanges of operating plant, the increase of the coolant pressure makes the purification temperature control difficult. In this paper, the controllability of the control dynamics of the purification system was analysed and the optimal parameters were proposed. To reduce the effects of the flow disturbance, the feedforward control structure was proposed and analysed.

  16. Blood purification and hemo- perfusion

    Institute of Scientific and Technical Information of China (English)


    The method of blood purification is a new overlapping frontierdiscipline which develops quickly in recent years. It helps overcoming many serious and complicated diseases, even including some incurable illnesses.

  17. Nanomechanical Water Purification Device Project (United States)

    National Aeronautics and Space Administration — Seldon Laboratories, LLC, proposes a lightweight, low-pressure water purification device that harnesses the unique properties of carbon nanotubes and will operate...

  18. The SELEX Air Purification System (United States)


    REPORT Final Report for the SELEX Air Purification System 14. ABSTRACT 16. SECURITY CLASSIFICATION OF: A new air purification technology ( SELEX ) was...developed and demonstrated. The SELEX system utilizes an array of electrospray wick aerosol sources for particle ionization and an electrostatic...precipitator for particle collection. The particle ionization process does not produce ozone and the SELEX technology provides a unique combination of

  19. One-step purification of twin-strep-tagged proteins and their complexes on strep-tactin resin cross-linked with bis(sulfosuccinimidyl) suberate (BS3). (United States)

    Ivanov, Konstantin I; Bašić, Marta; Varjosalo, Markku; Mäkinen, Kristiina


    Affinity purification of Strep-tagged fusion proteins on resins carrying an engineered streptavidin (Strep-Tactin) has become a widely used method for isolation of protein complexes under physiological conditions. Fusion proteins containing two copies of Strep-tag II, designated twin-Strep-tag or SIII-tag, have the advantage of higher affinity for Strep-Tactin compared to those containing only a single Strep-tag, thus allowing more efficient protein purification. However, this advantage is offset by the fact that elution of twin-Strep-tagged proteins with biotin may be incomplete, leading to low protein recovery. The recovery can be dramatically improved by using denaturing elution with sodium dodecyl sulfate (SDS), but this leads to sample contamination with Strep-Tactin released from the resin, making the assay incompatible with downstream proteomic analysis. To overcome this limitation, we have developed a method whereby resin-coupled tetramer of Strep-Tactin is first stabilized by covalent cross-linking with Bis(sulfosuccinimidyl) suberate (BS3) and the resulting cross-linked resin is then used to purify target protein complexes in a single batch purification step. Efficient elution with SDS ensures good protein recovery, while the absence of contaminating Strep-Tactin allows downstream protein analysis by mass spectrometry. As a proof of concept, we describe here a protocol for purification of SIII-tagged viral protein VPg-Pro from nuclei of virus-infected N. benthamiana plants using the Strep-Tactin polymethacrylate resin cross-linked with BS3. The same protocol can be used to purify any twin-Strep-tagged protein of interest and characterize its physiological binding partners.

  20. Purification and characterization of the Oligosaccharyl transferase

    Energy Technology Data Exchange (ETDEWEB)

    Kapoor, T.M.


    Oligosaccharyl transferase was characterized to be a glycoprotein with at least one saccharide unit that had a D-manno or D- glucopyranose configuration with unmodified hydroxy groups at C-3, C-4 and C-6, using a Concanavalin A affinity column. This afforded a 100 fold increase in the transferase purity in the solubilized microsomal sample and also removed over 90% of the microsomal proteins (the cytosolic ones being removed before solubilization). The detergent, N,N-Dimethyldodecylamine N-oxide (LDAO) was used for solubilization and it yielded a system compatible with the assay and the purification steps. An efficient method for detergent extraction without dilution of sample or protein precipitation was also developed.

  1. A colorimetric assay for cytokinin oxidase. (United States)

    Libreros-Minotta, C A; Tipton, P A


    A simple and rapid colorimetric assay for cytokinin oxidase is described. The assay is based on the formation of a Schiff base between the enzymatic reaction product 3-methyl-2-butenal and p-aminophenol. The assay is effective in the submicromolar concentration range and can be used in crude plant extracts as well as in more highly purified preparations.

  2. Screening of Indian medicinal plants for cytotoxic activity by Brine Shrimp Lethality (BSL assay and evaluation of their total phenolic content

    Directory of Open Access Journals (Sweden)

    Mahesh Biradi


    Full Text Available Objective: Plant-derived cytotoxic constituents and polyphenolic compounds have played an important role in the development of clinically useful anticancer agents. In this context, we have selected six Indian medicinal plants based on the literature claims and an attempt was made to evaluate the cytotoxic potential and total phenolic content (TPC of their methanol extracts and fractions. Materials and Methods: Six plants have been selected for the study, namely, Artemisia absinthium Linn. (Asteraceae, Oroxylum indicum (Linn. Vent. (Bignoniaceae, Heliotropium indicum Linn. (Boraginaceae, Amorphophallus sylvaticus (Roxb. Kunth. (Araceae, Mimosa pudica Linn. (Mimosaceae, and Premna serratifolia Linn. (Verbenaceae. Authenticated plant materials were subjected to extraction with methanol by cold maceration and hot percolation methods. The extracts were fractionated into four fractions (F1, F2, F3, and F4. Preliminary phytochemical investigation was carried out for all extracts and fractions. All extracts and their fractions were subjected to cytotoxicity screening by brine shrimp lethality (BSL bioassay. The plants with significant cytotoxicity were evaluated for TPC by using Folin-Ciocalteu reagent. Results: F1, F2, and F3 fractions of A. absinthium and P. serratifolia and F1 fraction of M. pudica have shown significant cytotoxicity (lethal concentration (LC 50 < 100 ppm compared with other fractions. F1, F2, and F3 fractions of A. absinthium show the LC 50 values 32.52, 14.27, and 24.02, respectively; F1, F2, and F3 of P. serratifolia show LC 50 values 7.61, 4.01, and 10.91 and same for F1 fraction of M. pudica was 34.82 μg/ml, respectively. TPC was found to be significantly higher (39.11 mg gallic acid equivalent (GAE/g in P. serratifolia compared with other two plants. Conclusion: The cytotoxicity screening system confirmed the proposed anticancer plants used by traditional healers and literature claims.

  3. Purification of equine Gc-globulin

    DEFF Research Database (Denmark)

    Houen, Gunnar; Pihl, Tina Holberg; Andersen, Pia Haubro;

    Objectives With the aim of producing antibodies for an equine Group specific component (Gc)-globulin assay, the protein was purified from normal equine plasma. Methods Equine Gc-globulin was purified from healthy horse plasma using ion exchange chromatography (Q-Sepharose, CM-Sepharose) and prepa......Objectives With the aim of producing antibodies for an equine Group specific component (Gc)-globulin assay, the protein was purified from normal equine plasma. Methods Equine Gc-globulin was purified from healthy horse plasma using ion exchange chromatography (Q-Sepharose, CM......-Sepharose) and preparative PAGE. Results Equine Gc-globulin has successfully been purified from healthy horse plasma and rabbits and mice are being immunized to produce specific antibodies. Conclusions Purification of equine Gc-globulin and the production of specific antibodies will make it possible to develop an assay...... for measuring Gc-globulin in horses. Studies in rodents and humans have shown that Gc-globulin is a multifunctional acute phase plasma protein, which removes actin from the blood by binding it and facilitating its clearance from the circulation by the liver. As such, Gc-globulin prevents hyper coagulation...

  4. High quality protein microarray using in situ protein purification

    Directory of Open Access Journals (Sweden)

    Fleischmann Robert D


    Full Text Available Abstract Background In the postgenomic era, high throughput protein expression and protein microarray technologies have progressed markedly permitting screening of therapeutic reagents and discovery of novel protein functions. Hexa-histidine is one of the most commonly used fusion tags for protein expression due to its small size and convenient purification via immobilized metal ion affinity chromatography (IMAC. This purification process has been adapted to the protein microarray format, but the quality of in situ His-tagged protein purification on slides has not been systematically evaluated. We established methods to determine the level of purification of such proteins on metal chelate-modified slide surfaces. Optimized in situ purification of His-tagged recombinant proteins has the potential to become the new gold standard for cost-effective generation of high-quality and high-density protein microarrays. Results Two slide surfaces were examined, chelated Cu2+ slides suspended on a polyethylene glycol (PEG coating and chelated Ni2+ slides immobilized on a support without PEG coating. Using PEG-coated chelated Cu2+ slides, consistently higher purities of recombinant proteins were measured. An optimized wash buffer (PBST composed of 10 mM phosphate buffer, 2.7 mM KCl, 140 mM NaCl and 0.05% Tween 20, pH 7.4, further improved protein purity levels. Using Escherichia coli cell lysates expressing 90 recombinant Streptococcus pneumoniae proteins, 73 proteins were successfully immobilized, and 66 proteins were in situ purified with greater than 90% purity. We identified several antigens among the in situ-purified proteins via assays with anti-S. pneumoniae rabbit antibodies and a human patient antiserum, as a demonstration project of large scale microarray-based immunoproteomics profiling. The methodology is compatible with higher throughput formats of in vivo protein expression, eliminates the need for resin-based purification and circumvents

  5. Angiogenesis Assays. (United States)

    Nambiar, Dhanya K; Kujur, Praveen K; Singh, Rana P


    Neoangiogenesis constitutes one of the first steps of tumor progression beyond a critical size of tumor growth, which supplies a dormant mass of cancerous cells with the required nutrient supply and gaseous exchange through blood vessels essentially needed for their sustained and aggressive growth. In order to understand any biological process, it becomes imperative that we use models, which could mimic the actual biological system as closely as possible. Hence, finding the most appropriate model is always a vital part of any experimental design. Angiogenesis research has also been much affected due to lack of simple, reliable, and relevant models which could be easily quantitated. The angiogenesis models have been used extensively for studying the action of various molecules for agonist or antagonistic behaviour and associated mechanisms. Here, we have described two protocols or models which have been popularly utilized for studying angiogenic parameters. Rat aortic ring assay tends to bridge the gap between in vitro and in vivo models. The chorioallantoic membrane (CAM) assay is one of the most utilized in vivo model system for angiogenesis-related studies. The CAM is highly vascularized tissue of the avian embryo and serves as a good model to study the effects of various test compounds on neoangiogenesis.

  6. Robotic high-throughput purification of affinity-tagged recombinant proteins. (United States)

    Wiesler, Simone C; Weinzierl, Robert O J


    Affinity purification of recombinant proteins has become the method of choice to obtain good quantities and qualities of proteins for a variety of downstream biochemical applications. While manual or FPLC-assisted purification techniques are generally time-consuming and labor-intensive, the advent of high-throughput technologies and liquid handling robotics has simplified and accelerated this process significantly. Additionally, without the human factor as a potential source of error, automated purification protocols allow for the generation of large numbers of proteins simultaneously and under directly comparable conditions. The delivered material is ideal for activity comparisons of different variants of the same protein. Here, we present our strategy for the simultaneous purification of up to 24 affinity-tagged proteins for activity measurements in biochemical assays. The protocol described is suitable for the scale typically required in individual research laboratories.

  7. 水葫芦生态净化工程对竺山湖底栖动物群落结构变化的影响%Effects of Ecological Purification Engineering of Planting Water Hyacinth on Macro-Benthos Community Structure

    Institute of Scientific and Technical Information of China (English)

    刘国锋; 韩士群; 何俊; 严少华; 周庆


    , which has the wide adaptability, large biomass, strong purification ability, especially the eutrophication water, after solving the mechanized harvesting, recycling use of terminal processing. The conventional ecological engineering practice is mainly in small water body or inland rivers, which has the obvious purification effects for no wind disturbance. But controlled planting the aquatic plants in large water body to purify the polluted water is still rare now. According to the instruction and requirements of Jiangsu Province, the ecological effects of planting 67 hectares water hyacinth (E. crassipens) in Zhushan Bay, Lake Taihu, which is one of the polluted lake water purification measures in Jiangsu Province and mainly planted by Jiangsu Academic Agricultural Science, on macro-benthos population and structure and benthos environment, were studied during 4~11 month in 2011 with consecutive surveys. Results indicated that average density mollusca (the main species were Bellamya aeruginosa) in far-planting, near-planting and planting area was 15.13、15.63、22.63 ind·m-2,respectively, and biomass was 17.00、17.60、25.50 g·m-2,respectively, showed that benthos biomass in planting area was higher than that the others. However, the average density and biomass of Oligochaeta (the main species were Limodrilus hoffmeisteri) and Chironomidae in planting area were lower than that outside of planting area, and it demonstrated that the benthic environment gradually improved after controlled planting the floating plants. It indicated that the ecological engineering management through planting water hyacinth couldn’t show the obvious purification effects of polluted water in a short time, especially in a shallow, wind disturbance of large lake, and it need long-term, lasting approached to reach the purifying goals. The index of Shannon-Weaver and Simpson indicated that water environment was severe polluted state. On the basis of the survey results, the large-area and high

  8. Garcinia lanceifolia Roxb; An Endemic Medicinal Plant of Assam Relieves Pain and Delays Nociceptive Response: An Assay for Its Analgesic and Anti-inflammatory Activity

    Directory of Open Access Journals (Sweden)

    Nilutpal Sharma Bora


    Full Text Available The stem bark of Garcinia lanceifolia, a lesser known species of the genus Garcinia, was extracted with methanol after pretreatment with petroleum ether. The resultant extract was evaluated for its analgesic and anti-inflammatory potential using various models. The assays were carried out on adult male Wistar Albino rats. The analgesic activity was assayed by hot plate method and tail flick assay using Morphine sulphate as standard drug at a dose of 5 mg/kg body weight and the results were expressed as mean increase in latency after drug administration ± SEM. The anti-inflammatory activity was evaluated by Carrageenan-induced rat paw oedema model using Diclofenac sodium as standard drug at a dose of 100 mg/kg body weight. The results were expressed in terms of mean increase in paw volume ± SEM. The extract of the stem bark was given in doses of 250 mg/kg and 500 mg/kg body weight. All the doses were administered per orally. Results reveal that Garcinia lanceifolia Roxb. posseses remarkable analgesic and anti-inflammatory activities.

  9. Assay of adenosine 3',5' cyclic monophosphate by stimulation of protein kinase: a method not involving radioactivity

    Energy Technology Data Exchange (ETDEWEB)

    Handa, A.K.; Bressan, R.A.


    In order to meet a need for a cAMP assay which is not subject to interference by compounds in plant extracts, and which is suitable for use on occasions separated by many /sup 32/P half-lives, an assay based on cAMP-dependent protein kinase has been developed which does not require the use of (..gamma..-/sup 32/P)ATP. Instead of measuring the cAMP-stimulated increase in the rate of transfer of (..gamma..-/sup 32/P) phosphate from (..gamma..-/sup 32/P)ATP to protein, the rate of loss of ATP from the reaction mixture is determined. The ATP remaining after the protein kinase reaction is assayed by ATP-dependent chemiluminescence of the firefly luciferin-luciferase system. Under conditions of the protein kinase reaction in which a readily measurable decrease in ATP concentration occurs, the logarithm of the concentration of ATP decreases in proportion to the cAMP concentration, i.e., the reaction can be described by the equation: (ATP) = (ATP)/sub 0/ e/sup -(cAMP)kt/. The assay based on this relationship can detect less than 1 pmol of cAMP. The levels of cAMP found with this assay after partial purification of the cAMP from rat tissue, algal cells, and the media in which the cells were grown agreed with measurements made by the cAMP binding-competition assay of Gilman, and the potein kinase stimulation assay based on transfer of (/sup 32/P) phosphate from (..gamma..-/sup 32/P)ATP to protein. All of the enzymes and chemicals required for the assay of cAMP by protein kinase catalyzed loss of ATP can be stored frozen for months, making the assay suitable for occasional use.

  10. 高等植物脂氧合酶活性测定方法的研究进展%Research Advances in the Assay Methods of the Lipoxygenase Activity in Higher Plants

    Institute of Scientific and Technical Information of China (English)

    李会容; 赵昶灵; 杨焕文; 邓建华; 毛自朝


    本文系统归纳了高等植物脂氧合酶活性检测方法的研究进展.常规检测方法可分为两类:一类基于检测脂氧合酶催化反应产生氢过氧化物的速度,包括胡萝卜素脱色法、Fe(CNS)3显色法、标准分光光度法、2-硫代巴比妥酸显色法、碘化钾-淀粉法、二甲苯酚橙法;另一类基于检测反应中底物摄取氧的速度,包括量压法、氧电极法、亚甲基蓝染色法.此外还有酶联免疫法、超弱发光发射光谱法以及放射性同位素标记法.其中,标准分光光度法和氧电极法为脂氧合酶活性检测的标准方法.本文可为特定高等植物脂氧合酶活性的高效、准确测定提供参考.%The research advances in the assay methods of the lipoxygenase activity in higher plants were systematically summarized in this review. The normal assay methods involve two categories. One is based on the rate determination of the hydroperoxide produced in the lipoxygenase-catalyzed reactions, including carotene bleaching, Fe(CNS)3 coloration, standard spectrophotometric, 2-thiobarbituric acid coloration,potassium iodide-starch and xylenol orange methods. The other is based on the determination of the oxygen-absorbing rate of the substrates in the lipoxygenase-catalyzed reactions, including manometric, oxygen electrode and methylene blue staining methods. Other assay method includes enzyme linked immunosorbent assay, superweak luminescence emission spectrum assay and radioisotopic labelling method. Among all of the above methods, the standard spectrophotometric and the oxygen electrode methods are both regarded as the standard ones. This paper could provide a reference for the efficient and accurate determination of the lipoxygenase activity in the specific higher plants.

  11. Study on the Purification and Tolerance Ability of Bonsai Aloe and Snake Plant to Formaldehyde in Indoor Air%盆栽芦荟和虎皮兰对室内空气甲醛净化及耐受能力的研究

    Institute of Scientific and Technical Information of China (English)

    郭铭伟; 龚萍; 陶荣琴; 殷姿; 梁维君


    目的 研究及比较盆栽芦荟和虎皮兰对室内空气甲醛的净化及耐受能力. 方法 选择芦荟、虎皮兰两种常见、易护理且经济的盆栽植物进行三个方面的实验,即植物吸收甲醛能力的判断、不同植物吸收甲醛能力大小的比较、植物对甲醛耐受能力的研究.测得实验前后甲醛的变化量来反映植物对甲醛的净化效果,通过观察实验组植物叶片是否出现斑点、叶片枯萎、叶片坏死情况来反映绿色植物对甲醛的耐受性. 结果 实验组的甲醛浓度比对照组的甲醛浓度明显降低(P<0.01),且虎皮兰组甲醛浓度低于芦荟组(P<0.05);实验组叶片斑点增加、叶尖变黄、叶片变黄、叶片枯死等发生情况均高于对照组(P<0.05),且对虎皮兰的损伤大于芦荟(P<0.05). 结论 芦荟和虎皮兰均能净化室内空气污染物甲醛;芦荟和虎皮兰净化甲醛的能力有差异,虎皮兰比芦荟净化甲醛的能力强;甲醛对盆栽绿色植物有损伤,虎皮兰比芦荟的耐受性低.%Objective To study and compare the purification and tolerance ability of Bonsai Aloe and Snake Plant to formaldehyde in indoor air. Methods Bonsai Aloe and Snake Plant were studied and compared on their abilities to absorb and toi-erate formaldehyde in indoor air. Their purification effects were studied by detecting the formaldehyde in indoor air before and after the plants were placed in the room, with a room without plant set as the control. Their tolerance ability to formaldehyde was determined by observing the changes of the leaves, such as appearing of spots, blight, and necrosis. Results The formaldehyde concentrations in the indoor air of the experimental groups after plants placement were significantly lower than that of the control group (P<0.01), and the concentration in the Snake Plant group was statistically lower than that in the Bonsai Aloe group (P<0. 05). More spotted leaves, yellow leaves and necrotic leaves

  12. Water Purification Systems (United States)


    Clearwater Pool Technologies employs NASA-developed silver/copper ionization to purify turtle and dolphin tanks, cooling towers, spas, water recycling systems, etc. The pool purifier consists of a microcomputer to monitor water conditions, a pair of metallic electrodes, and a rheostat controller. Ions are generated by passing a low voltage current through the electrodes; the silver ions kill the bacteria, and the copper ions kill algae. This technology has found broad application because it offers an alternative to chemical disinfectants. It was originally developed to purify water on Apollo spacecraft. Caribbean Clear has been using NASA's silver ionization technology for water purification for more than a decade. Two new products incorporate advancements of the basic technology. One is the AquaKing, a system designed for areas with no source of acceptable drinking water. Another is the Caribbean Clear Controller, designed for commercial pool and water park applications where sanitizing is combined with feedback control of pH and an oxidizer, chlorine or bromine. The technology was originally developed to purify water on Apollo spacecraft.

  13. Protocol for Initial Purification of Bacteriocin (United States)


    lysate/extract preparation , column purification, and a desalting. The peptide was tracked throughout the process using a soft agar overlay activity... PREPARATION SOLUBLE EXTRACTS COLUMN CHROMATOGRAPHY BACTERIA PURIFICATION CHARACTERIZATION...3  6.1  Preparation of Target Bacteria

  14. Purification and identification of antibacterial phenolics from Tripodanthus acutifolius leaves. (United States)

    Soberón, J R; Sgariglia, M A; Sampietro, D A; Quiroga, E N; Sierra, M G; Vattuone, M A


    To perform an activity-guided purification, identification and quantification of antibacterial compounds from Tripodanthus acutifolius infusion. To validate the antibacterial activity of purified substances. Bioautographic methods were employed as screening assays for purifying bioactive substances. Purification procedures included sephadex LH-20 column chromatography and reverse phase HPLC. Identification was achieved by spectroscopic methods (UV-Vis, MS, NMR and polarimetry) and chromatographic assays (paper chromatography and HPLC). Antibacterial activity was studied by microdilution, colony count and photometric assays, Sytox green stain and transmission electron microscopy (TEM). Four glycoflavonoids (rutin, nicotiflorin, hyperoside and isoquercitrin) and an unusual phenylbutanoid glycoside (tripodantoside) were purified and identified. Tripodantoside was found at 6.59 +/- 0.82 g per 100 g of dry leaves. The flavonoids showed bactericidal effect at a concentration of 4 mg ml(-1) against Staphylococcus aureus and Pseudomonas aeruginosa strains from American Type Culture Collection, while tripodantoside was almost four times more active than those compounds, with a minimum bactericidal concentration = 1.024 mg ml(-1) against these strains. Tripodantoside aglycone showed bacteriolytic effects on the assayed strains, causing evident damages on cell wall and membrane, while tripodantoside did not exhibit those effects. The antibacterial activity of T. acutifolius infusion would be partially attributed to the purified glycoflavonoids and mainly to tripodantoside. The high extraction yield and the antibacterial activity exhibited by tripodantoside makes this chemical structure of interest to support further studies dealing with chemical modifications to increase the antibacterial activity or to seek another activities.

  15. Development and comparison of membrane methods for improving the performance of municipal waster water purification by combination of activation biology and microfiltration and construction and optimisation of an experimental plant; Entwicklung und Vergleich von Membranverfahren zur Leistungssteigerung der kommunalen Abwasserreinigung durch die Kombination von Belebungsbiologie und Mikrofiltration sowie Bau und Optimierung einer technischen Versuchsanlage

    Energy Technology Data Exchange (ETDEWEB)

    Finner, M.


    The goal of the present project was to develop a membrane-based activation method, optimise it on an experimental plant and operate this plant alongside a conventional municipal water treatment plant over a one-year period. An important part of the project consisted in making a technical comparison between conventional water purification and the membrane-based activation method. The goal was to optimise the membrane-based activation method to a point where its use in place of conventional methods based on activation biology could be considered promising also in economic terms. [German] In dem Projekt war es vorgesehen, ein Membranbelebungsverfahren zu entwickeln, durch den Betrieb einer technischen Anlage zu optimieren und diese parallel zu einer konventionellen kommunalen Klaeranlage ueber einen Zeitraum von einem Jahr zu betreiben. Ein wesentlicher Projektbestandteil war der technische Vergleich der konventionellen Abwasserreinigung mit dem Membranbelebungsverfahren. Ziel war es, das Membranbelebungsverfahren technisch soweit zu optimieren, dass sein Einsatz als Alternative zu konventionellen Belebungsbiologien kuenftig auch unter wirtschaftlichen Bedingungen als erfolgversprechend gelten kann. (orig.)

  16. Rapid purification of recombinant histones.

    Directory of Open Access Journals (Sweden)

    Henrike Klinker

    Full Text Available The development of methods to assemble nucleosomes from recombinant histones decades ago has transformed chromatin research. Nevertheless, nucleosome reconstitution remains time consuming to this day, not least because the four individual histones must be purified first. Here, we present a streamlined purification protocol of recombinant histones from bacteria. We termed this method "rapid histone purification" (RHP as it circumvents isolation of inclusion bodies and thereby cuts out the most time-consuming step of traditional purification protocols. Instead of inclusion body isolation, whole cell extracts are prepared under strongly denaturing conditions that directly solubilize inclusion bodies. By ion exchange chromatography, the histones are purified from the extracts. The protocol has been successfully applied to all four canonical Drosophila and human histones. RHP histones and histones that were purified from isolated inclusion bodies had similar purities. The different purification strategies also did not impact the quality of octamers reconstituted from these histones. We expect that the RHP protocol can be readily applied to the purification of canonical histones from other species as well as the numerous histone variants.

  17. Advanced purification of petroleum refinery wastewater by catalytic vacuum distillation. (United States)

    Yan, Long; Ma, Hongzhu; Wang, Bo; Mao, Wei; Chen, Yashao


    In our work, a new process, catalytic vacuum distillation (CVD) was utilized for purification of petroleum refinery wastewater that was characteristic of high chemical oxygen demand (COD) and salinity. Moreover, various common promoters, like FeCl(3), kaolin, H(2)SO(4) and NaOH were investigated to improve the purification efficiency of CVD. Here, the purification efficiency was estimated by COD testing, electrolytic conductivity, UV-vis spectrum, gas chromatography-mass spectrometry (GC-MS) and pH value. The results showed that NaOH promoted CVD displayed higher efficiency in purification of refinery wastewater than other systems, where the pellucid effluents with low salinity and high COD removal efficiency (99%) were obtained after treatment, and the corresponding pH values of effluents varied from 7 to 9. Furthermore, environment estimation was also tested and the results showed that the effluent had no influence on plant growth. Thus, based on satisfied removal efficiency of COD and salinity achieved simultaneously, NaOH promoted CVD process is an effective approach to purify petroleum refinery wastewater. Copyright 2010 Elsevier B.V. All rights reserved.

  18. Assays of the production of harmful substances by genetically modified oilseed rape (Brassica napus L.) plants in accordance with regulations for evaluating the impact on biodiversity in Japan. (United States)

    Asanuma, Yoko; Jinkawa, Tomoe; Tanaka, Hidenori; Gondo, Takahiro; Zaita, Norihiro; Akashi, Ryo


    Environmental risk assessment of transgenic crops is implemented under the Cartagena Protocol domestic law in accordance with guidelines for implementing the assessment established by the Ministry of Agriculture, Forestry and Fisheries (MAFF) and the Ministry of Environment (MOE) in Japan. Environmental risk assessments of transgenic crops are implemented based on the concept of 'substantial equivalence' to conventional crops. A unique requirement in Japan to monitor the production of harmful substances, or allelochemicals, is unparalleled in other countries. The potential for allelochemicals to be secreted from the roots of transgenic crops to affect other plants or soil microflora or for substances in the plant body to affect other plants after dying out must be evaluated. We evaluated the allelopathic potential of seven transgenic oilseed rape (Brassica napus L.) lines that express glufosinate tolerance in terms of substantial equivalence to conventional oilseed rape lines, and established evaluation methods. Our results indicate no potential production of allelochemicals for any of the seven transgenic oilseed rape lines compared with conventional oilseed rape lines.

  19. CTAB-silica Method for DNA Extraction and Purification from Castanea mollissima and Ginkgo biloba

    Institute of Scientific and Technical Information of China (English)

    Shen Yongbao; Shi Jisen


    A new method CTAB-silica for DNA extraction and purification from the leaves and buds of Castanea mollissima and Ginkgo biloba was tested. The method is based on the silica-based purification protocol developed by Boom et al. (1990). By modifying the protocol, plant genome DNA could be extracted easily from dormant buds, mature leaves, and other parts of plant. Our results showed that the purified DNA was of high purity and could be analyzed by PCR. Furthermore, this CTAB-silica method took much less time for a successful DNA purification process compared to the traditional methods (CTAB and SDS). By our method, the suitable DNA can be extracted and purified from over 10 plant samples by one person in an hour.


    Directory of Open Access Journals (Sweden)

    Azam Khodashenas Pelko


    Full Text Available The Jainism emphasizes three major teachings about the purification of the soul (jiva, Ahimsa, Aparigrapha and anekantwad. Jainism, The focus of this religion has been purification of the soul by means of right conduct, right faith and right knowledge. The ultimate goal of Hinduism is Moksha or liberation (total freedom. In Hinduism, purification of the soul is a goal that one must work to attain. The Buddhism is the science of pursuing the aim of making the human mind perfect, and of purifying the human soul. The knowledge of purifying of the soul and softening of the hearts is as essential for human. They having the correct motivations means purifying our souls from hypocrisy, caprice, and heedlessness. The primary goal of Taoism may be described as the mystical intuition of the Tao, which is the way, the undivided unity, and the ultimate Reality. According to the Christianity access to truth cannot be conceived without purity of the soul

  1. Assay and heterologous expression in Pichia pastoris of plant cell wall type-II membrane anchored glycosyltransferases

    DEFF Research Database (Denmark)

    Petersen, Bent Larsen; Egelund, Jack; Damager, Iben


    Two Arabidopsis xylosyltransferases, designated RGXT1 and RGXT2, were recently expressed in Baculovirus transfected insect cells and by use of the free sugar assay shown to catalyse transfer of D-xylose from UDP-α-D-xylose to L-fucose and derivatives hereof. We have now examined expression of RGXT1...... incubated with 0.5 M L-fucose and UDP-D-xylose all four RGXT1 and RGXT2 variants catalyzed transfer of D-xylose onto L-fucose with estimated turnover numbers between 0.15 and 0.3 sec-1, thus demonstrating that a free C-terminus is not required for activity. N- and O-glycanase treatment resulted...

  2. Research on landscape water purification by seeds of natural plant Moringa oleifera%天然植物辣木籽对水体净化作用的研究

    Institute of Scientific and Technical Information of China (English)

    张饮江; 王聪; 刘晓培; 董悦; 李岩; 文晓峰; 马海峰


    This paper made a comparative study on purifying effect among natural water purification a-gent Moringa seed, commonly-used chemical agent alum and polymeric aluminum chloride. The results indicate that Moringa seed is a very good natural water purification agent, and its purification efficiency is comparable to that of the polymeric aluminum chloride. The optimal dose of Moringa seed is 100 mg/L in purifying water. The crude extraction of Moringa seed has a good purifying effect but it increases organic matter content. Through the treatment of Moringa seed by using different times distilled water and NaCl solution, it is found that Moringa seed extracted by 10 times NaCl solution is the best to reduce turbidity effect, while the CODMn index does not increase. The turbidity reduction effect is on the same level when using different salt solution to extract Moringa seed. Therefore the seed of Moringa oleifera has potential value as a natural purifying agent.%文章对辣木籽天然净水剂与常用化学净水剂明矾、聚合氯化铝等净水效果进行比较研究,结果表明:辣木籽有较明显的净水效果,净水效率与聚合氯化铝相当;确定辣木籽净水最佳剂量为100mg/L;辣木籽粗提取液有良好净水效果,但水体中有机物含量有所增加;利用不同倍数蒸馏水与NaCl溶液对辣木籽进行处理,10倍NaCl溶液提取辣木籽,降低浊度的效果明显,同时高锰酸盐指数并未增加,采用不同盐溶液提取辣木籽,对水体浊度去除效果相当.因此,辣木籽作为天然净水剂具有较好的潜在应用价值.

  3. In vitro antioxidant and antiproliferative activities of methanolic plant part extracts of Theobroma cacao. (United States)

    Baharum, Zainal; Akim, Abdah Md; Taufiq-Yap, Yun Hin; Hamid, Roslida Abdul; Kasran, Rosmin


    The aims of this study were to determine the antioxidant and antiproliferative activity of the following Theobroma cacao plant part methanolic extracts: leaf, bark, husk, fermented and unfermented shell, pith, root, and cherelle. Antioxidant activity was determined using 2,2-diphenyl-2-picrylhydrazyl (DPPH), thiobarbituric acid-reactive substances (TBARS), and Folin-Ciocalteu assays; the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium (MTT) assay was used to determine antiproliferative activity. The root extract had the highest antioxidant activity; its median effective dose (EC50) was 358.3±7.0 µg/mL and total phenolic content was 22.0±1.1 g GAE/100 g extract as compared to the other methanolic plant part extracts. Only the cherelle extract demonstrated 10.4%±1.1% inhibition activity in the lipid peroxidation assay. The MTT assay revealed that the leaf extract had the highest antiproliferative activity against MCF-7 cells [median inhibitory concentration (IC50)=41.4±3.3 µg/mL]. Given the overall high IC50 for the normal liver cell line WRL-68, this study indicates that T. cacao methanolic extracts have a cytotoxic effect in cancer cells, but not in normal cells. Planned future investigations will involve the purification, identification, determination of the mechanisms of action, and molecular assay of T. cacao plant extracts.

  4. In Vitro Antioxidant and Antiproliferative Activities of Methanolic Plant Part Extracts of Theobroma cacao

    Directory of Open Access Journals (Sweden)

    Zainal Baharum


    Full Text Available The aims of this study were to determine the antioxidant and antiproliferative activity of the following Theobroma cacao plant part methanolic extracts: leaf, bark, husk, fermented and unfermented shell, pith, root, and cherelle. Antioxidant activity was determined using 2,2-diphenyl-2-picrylhydrazyl (DPPH, thiobarbituric acid-reactive substances (TBARS, and Folin-Ciocalteu assays; the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium (MTT assay was used to determine antiproliferative activity. The root extract had the highest antioxidant activity; its median effective dose (EC50 was 358.3 ± 7.0 µg/mL and total phenolic content was 22.0 ± 1.1 g GAE/100 g extract as compared to the other methanolic plant part extracts. Only the cherelle extract demonstrated 10.4% ± 1.1% inhibition activity in the lipid peroxidation assay. The MTT assay revealed that the leaf extract had the highest antiproliferative activity against MCF-7 cells [median inhibitory concentration (IC50 = 41.4 ± 3.3 µg/mL]. Given the overall high IC50 for the normal liver cell line WRL-68, this study indicates that T. cacao methanolic extracts have a cytotoxic effect in cancer cells, but not in normal cells. Planned future investigations will involve the purification, identification, determination of the mechanisms of action, and molecular assay of T. cacao plant extracts.

  5. Bioassay of Estrogenic Activity of Effluent and Influent in a Farm Wastewater Treatment Plant Using an in vitro Recombinant Assay with Yeast Cells

    Institute of Scientific and Technical Information of China (English)



    Objective Environmental estrogens at an elevated concentration are known to produce adverse effects on human and animal life. However, the majority of researches have been focused on ndustrial discharges, while the impact of livestock wastes as a source of endocrine disrupters in aquatic environments has been rarely elucidated. In order to investigate the contribution of environmental estrogens from livestock, the estrogenic activity in water samples from a farm wastewater treatment plant was analyzed by a recombinant yeast screening method. Methods The extracts prepared from 15 selected water samples from the farm wastewater treatment plant, among which 6 samples were from pre-treatment section (influents) and 9 from post-treatment section (effluents), were analyzed for estrogenic activity by cellar bioassay. Yeast cells transfected with the expression plasmid of human estrogen receptor and the Lac Z reporter plasmid encoding β-galactossidase, were used to measure the estrogen-like compounds in the farm wastewater treatment plant. Results The wastewater samples from influents showed a higher estrogenic potency than the effluent samples showing a low induction of β-galactossidase relative to solvent control condition. By comparison with a standard curve for 1713-estradiol (E2), estrogenic potency in water samples from the influents was calculated as E2-equivalent and ranged from 0.1 to 150 pM E2-equivalent. The estrogenic potency in water samples from the effluents was significantly lower than that in the influents, and 7 water samples had less detectable limit in the total of 9 samples. Conclusion Yeast bioassay of estrogenic activity in most of the samples from the farm wastewater after disposal by traditional sewage treatment showed negative results.

  6. pSiM24 is a novel versatile gene expression vector for transient assays as well as stable expression of foreign genes in plants. (United States)

    Sahoo, Dipak Kumar; Dey, Nrisingha; Maiti, Indu Bhushan


    We have constructed a small and highly efficient binary Ti vector pSiM24 for plant transformation with maximum efficacy. In the pSiM24 vector, the size of the backbone of the early binary vector pKYLXM24 (GenBank Accession No. HM036220; a derivative of pKYLX71) was reduced from 12.8 kb to 7.1 kb. The binary vector pSiM24 is composed of the following genetic elements: left and right T-DNA borders, a modified full-length transcript promoter (M24) of Mirabilis mosaic virus with duplicated enhancer domains, three multiple cloning sites, a 3'rbcsE9 terminator, replication functions for Escherichia coli (ColE1) and Agrobacterium tumefaciens (pRK2-OriV) and the replicase trfA gene, selectable marker genes for kanamycin resistance (nptII) and ampicillin resistance (bla). The pSiM24 plasmid offers a wide selection of cloning sites, high copy numbers in E. coli and a high cloning capacity for easily manipulating different genetic elements. It has been fully tested in transferring transgenes such as green fluorescent protein (GFP) and β-glucuronidase (GUS) both transiently (agro-infiltration, protoplast electroporation and biolistic) and stably in plant systems (Arabidopsis and tobacco) using both agrobacterium-mediated transformation and biolistic procedures. Not only reporter genes, several other introduced genes were also effectively expressed using pSiM24 expression vector. Hence, the pSiM24 vector would be useful for various plant biotechnological applications. In addition, the pSiM24 plasmid can act as a platform for other applications, such as gene expression studies and different promoter expressional analyses.

  7. pSiM24 is a novel versatile gene expression vector for transient assays as well as stable expression of foreign genes in plants.

    Directory of Open Access Journals (Sweden)

    Dipak Kumar Sahoo

    Full Text Available We have constructed a small and highly efficient binary Ti vector pSiM24 for plant transformation with maximum efficacy. In the pSiM24 vector, the size of the backbone of the early binary vector pKYLXM24 (GenBank Accession No. HM036220; a derivative of pKYLX71 was reduced from 12.8 kb to 7.1 kb. The binary vector pSiM24 is composed of the following genetic elements: left and right T-DNA borders, a modified full-length transcript promoter (M24 of Mirabilis mosaic virus with duplicated enhancer domains, three multiple cloning sites, a 3'rbcsE9 terminator, replication functions for Escherichia coli (ColE1 and Agrobacterium tumefaciens (pRK2-OriV and the replicase trfA gene, selectable marker genes for kanamycin resistance (nptII and ampicillin resistance (bla. The pSiM24 plasmid offers a wide selection of cloning sites, high copy numbers in E. coli and a high cloning capacity for easily manipulating different genetic elements. It has been fully tested in transferring transgenes such as green fluorescent protein (GFP and β-glucuronidase (GUS both transiently (agro-infiltration, protoplast electroporation and biolistic and stably in plant systems (Arabidopsis and tobacco using both agrobacterium-mediated transformation and biolistic procedures. Not only reporter genes, several other introduced genes were also effectively expressed using pSiM24 expression vector. Hence, the pSiM24 vector would be useful for various plant biotechnological applications. In addition, the pSiM24 plasmid can act as a platform for other applications, such as gene expression studies and different promoter expressional analyses.

  8. In vitro genotoxicity and cytotoxicity of five new chemical compounds of plant origin by means of the human lymphocyte micronucleus assay. (United States)

    Scarpato, R; Pistelli, L; Bertoli, A; Nieri, E; Migliore, L


    The micronucleus test in human peripheral lymphocytes is widely used in toxicology for the assessment of the genotoxic profile of chemical compounds of environmental concern. The aim of this study was to evaluate the genotoxic, cytotoxic and antimitotic activity of five new compounds isolated from Prunus africana Hook or from Bupleurum fruticosum L. The experiments were conducted only in vitro. Results showed that none of the plant extracts, tested over a wide range of concentrations, increased the frequency of micronuclei. Only compounds 2 and 5 were found to be toxic for phytohaemagglutinin-stimulated lymphocytes at the maximum dose used. reserved.

  9. Neurotrophic factor - Characterization and partial purification (United States)

    Popiela, H.; Ellis, S.


    Recent evidence suggests that neurotrophic activity is required for the normal proliferation and development of muscle cells. The present paper reports a study of the purification and characterization of a neurotrophic factor (NTF) from adult chicken ischiatic-peroneal nerves using two independent quantitative in vitro assay systems. The assays were performed by the measurement of the incorporation of tritiated thymidine or the sizes of single-cell clones by chick muscle cells grown in culture. The greatest amount of neutrotrophic activity is found to be extracted at a pH of 8; aqueous suspensions of the activity are stable to long-term storage at room temperature. The specific activity of the substance is doubled upon precipitation with ammonium sulfate or after gel filtration, and increase 4 to 5 fold after salt gradient elution from DEAE cellulose columns. The active fraction obtained after gel filtration and rechromatography on DEAE cellulose exhibits a 7 to 10-fold increase in specific activity. Electrophoresis of the most highly purified material yields a greatly concentrated band at around 80,000 daltons. Although NTF is purified almost 10-fold as indicated by the increase in specific activity, the maximum activity of the partially purified material is greatly reduced, possibly due to a requirement for a cofactor for the expression of maximum activity.

  10. Hydrogen purification by periodic adsorption

    Energy Technology Data Exchange (ETDEWEB)

    Barg, Christian; Secchi, Argimiro R.; Trierweiler, Jorge O. [Rio Grande do Sul Univ., Porto Alegre, RS (Brazil). Dept. de Engenharia Quimica]. E-mail:;;


    The periodic adsorption processes have been widely used for industrial applications, mainly because it spends less energy than the usual gas separation processes, like the cryogenic distillation. The largest commercial application of periodic adsorption processes is the pressure swing adsorption (PSA) applied to hydrogen purification. Although its wide use in the chemical and petrochemical industry, there are no reports in the open literature about complete modeling studies of a complex commercial unit, with multiple adsorbents and multiple beds and several feed components. This study has as objective the modeling, optimization and dynamical analysis of an industrial PSA unit for hydrogen purification. (author)

  11. Upscaled CTAB-based DNA extraction and real-time PCR assays for Fusarium culmorum and F. graminearum DNA in plant material with reduced sampling error. (United States)

    Brandfass, Christoph; Karlovsky, Petr


    Fusarium graminearum Schwabe (Gibberella zeae Schwein. Petch.) and F. culmorum W.G. Smith are major mycotoxin producers in small-grain cereals afflicted with Fusarium head blight (FHB). Real-time PCR (qPCR) is the method of choice for species-specific, quantitative estimation of fungal biomass in plant tissue. We demonstrated that increasing the amount of plant material used for DNA extraction to 0.5-1.0 g considerably reduced sampling error and improved the reproducibility of DNA yield. The costs of DNA extraction at different scales and with different methods (commercial kits versus cetyltrimethylammonium bromide-based protocol) and qPCR systems (doubly labeled hybridization probes versus SYBR Green) were compared. A cost-effective protocol for the quantification of F. graminearum and F. culmorum DNA in wheat grain and maize stalk debris based on DNA extraction from 0.5-1.0 g material and real-time PCR with SYBR Green fluorescence detection was developed.


    Directory of Open Access Journals (Sweden)

    Yu. P. Sedlukho


    Full Text Available The paper considers problems and features of biochemical removal of hydrogen sulfide from ground water. The analysis of existing methods for purification of ground water from hydrogen sulfide has been given in the paper. The paper has established shortcomings of physical and chemical purification of ground water. While using aeration methods for removal of hydrogen sulfide formation of colloidal sulfur that gives muddiness and opalescence to water occurs due to partial chemical air oxidation. In addition to this violation of sulfide-carbonate equilibrium taking place in the process of aeration due to desorption of H2S and CO2, often leads to clogging of degasifier nozzles with formed CaCO3 that causes serious operational problems. Chemical methods require relatively large flow of complex reagent facilities, storage facilities and transportation costs.In terms of hydrogen sulfide ground water purification the greatest interest is given to the biochemical method. Factors deterring widespread application of the biochemical method is its insufficient previous investigation and necessity to execute special research in order to determine optimal process parameters while purifying groundwater of a particular water supply source. Biochemical methods for oxidation of sulfur compounds are based on natural biological processes that ensure natural sulfur cycle. S. Vinogradsky has established a two-stage mechanism for oxidation of hydrogen sulfide with sulfur bacteria (Beggiatoa. The first stage presupposes oxidation of hydrogen sulphide to elemental sulfur which is accumulating in the cytoplasm in the form of globules. During the second stage sulfur bacteria begin to oxidize intracellular sulfur to sulfuric acid due to shortage of hydrogen sulfide.The paper provides the results of technological tests of large-scale pilot plants for biochemical purification of groundwater from hydrogen sulfide in semi-industrial conditions. Dependences of water quality

  13. Assays of polychlorinated biphenyl congeners and co-contaminated heavy metals in the transgenic Arabidopsis plants carrying the recombinant guinea pig aryl hydrocarbon receptor-mediated β-glucuronidase reporter gene expression system. (United States)

    Shimazu, Sayuri; Ohta, Masaya; Ohkawa, Hideo; Ashida, Hitoshi


    The transgenic Arabidopsis plant XgD2V11-6 carrying the recombinant guinea pig (g) aryl hydrocarbon receptor (AhR)-mediated β-glucuronidase (GUS) reporter gene expression system was examined for assay of polychlorinated biphenyl (PCB) congeners and co-contaminated heavy metals. When the transgenic Arabidopsis plants were treated with PCB126 (toxic equivalency factor; TEF: 0.1) and PCB169 (TEF: 0.03), the GUS activity of the whole plants was increased significantly. After treatment with PCB80 (TEF: 0), the GUS activity was nearly the same level as that treated with 0.1% dimethylsulfoxide (DMSO) as a vehicle control. After exposure to a 1:1 mixture of PCB126 and PCB169, the GUS activity was increased additively. However, after exposure to a mixture of PCB126 and PCB80, the GUS activity was lower than that of the treatment with PCB126 alone. Thus, PCB80 seemed to be an antagonist towards AhR. When the transgenic plants were treated with each of the heavy metals Fe, Cu, Zn, Cd and Pb together with PCB126, Cd and Pb increased the PCB126-induced GUS activity. On the other hand, Fe, Cu and Zn did not affect the PCB126-induced GUS activity. In the presence of the biosurfactant mannosylerythritol lipid-B (MEL-B) and the carrier protein bovine serum albumin (BSA), the PCB126-induced GUS activity was increased, but the Cd-assisted PCB126-induced GUS activity was not affected. Thus, MEL-B and BSA seemed to increase uptake and transport of PCB126, respectively.

  14. A new QRT-PCR assay designed for the differentiation between elements provided from Agrobacterium sp. in GMOs plant events and natural Agrobacterium sp. bacteria. (United States)

    Nabi, Nesrine; Chaouachi, Maher; Zellama, Mohamed Salem; Ben Hafsa, Ahmed; Mrabet, Besma; Saïd, Khaled; Fathia, Harzallah Skhiri


    The question asked in the present work was how to differentiate between contamination of field samples with and GM plants contained sequences provided from this bacterium in order to avoid false positives in the frame of the detection and the quantification of GMO. For this, new set of primers and corresponding TaqMan Minor Groove Binder (MGB) probes were designed to target Agrobacterium sp. using the tumor-morphology-shooty gene (TMS1). Final standard curves were calculated for each pathogen by plotting the threshold cycle value against the bacterial number (log (colony forming units) per milliliter) via linear regression. The method designed was highly specific and sensitive, with a detection limit of 10CFU/ml. No significant cross-reaction was observed. Results from this study showed that TaqMan real-time PCR, is potentially an effective method for the rapid and reliable quantification of Agrobacterium sp. in samples containing GMO or non GMO samples.

  15. Demonstration project of an autonomous hybrid power supply with computer supported energy management for the purification plant Koerkwitz/Riebnitz-Damgarten (Baltic Sea area, Germany). Subproject: PV/biogas. Final report; Demonstrationsvorhaben zur objektorientierten hybriden Stromversorgung des Klaerwerkes Koerkwitz/Riebnitz-Damgarten mit DV-gestuetztem Energiemanagement. Teilprojekt: PV/Biogas. Schlussbericht

    Energy Technology Data Exchange (ETDEWEB)

    Krausen, E.; Neuhaeusser, G.


    For the purification plant Koerkwitz (Baltic Sea area, Germany) an autonomous hybrid power supply system was developed and installed using the regenerative energy sources sun and wind. The realized energy management is furthermore prepared for the additional installation of a bio-gas plant. For the final realization stage of self-supporting portion of more than 80% is expected. Surplus energy is fed to the public grid. Main components of the installed plant are a 250 kWp solar generator and a 300 kW wind converter. For the energy conversion three in parallel connected inverters are installed in a master/slave mode. An efficiency optimization is obtained by Maximum-Power-Point controlling and automatic switching on and off under partial load operation. The whole process control is performed by an energy management system which consists of a command caculator, central compact programmable controllers and local data acquisition devices for subsystems. The plant was started up in September, 1993 and is operating since that time without problems. (orig.) [Deutsch] Fuer das Klaerwerk Koerkwitz/Riebnitz-Damgarten ist eine objektorientierte hybride Eigenstromversorgungsanlage fuer die Nutzung der regenerativen Energiequellen Sonne und Wind installiert worden. Das Energiekonzept sowie das realisierte Energiemanagement sind fuer eine Erweiterung um eine Biogas-Anlage bereits vorbereitet. Im Endausbau wird dann eine Versorgungs-Autarkie von ueber 80% erwartet. Zeitweise ueberschuessige Energie wird ins oeffentliche Netz eingespeist. Die wesentlichen Komponenten der installierten Anlage sind ein 250 kWp Solargenerator sowie ein 300 kW Windkonverter. Die Aufbereitung des vom Solargenerator erzeugten Stroms folgt ueber drei parallel geschaltete Wechselrichter im Master/Slave-Betrieb, fuer die eine Wirkungsgradoptimierung durch MPP-Regelung und automatische leistungsabhaengige Zu- und Abschaltung im Teillastbereich erreicht wird. Die gesamte Prozessfuehrung wird von einem

  16. Purification and properties of dialkylfluorophosphatase

    NARCIS (Netherlands)

    Cohen, J.A.; Warringa, M.G.P.J.


    1. 1. Zone electrophoresis on starch columns of purified preparations of fluorophosphatase resulted in a further purification. The preparations thus obtained differed in various respects from the cruder ones so far described. 2. 2. In the course of this electrophoresis fractions were obtained, which

  17. Quantum entanglement purification in cavities

    CERN Document Server

    Romero, J L; Saavedra, C; Retamal, J C


    A physical implementation of an entanglement purification protocol is studied using a cavity quantum electrodynamic based proposal, where, the quantum information is stored in quantum field sates inside cavities. Also a procedure is given for quantifying the degree of entanglement between quantum fields. (Author)

  18. Bioinspired Materials for Water Purification

    Directory of Open Access Journals (Sweden)

    Alfredo Gonzalez-Perez


    Full Text Available Water scarcity issues associated with inadequate access to clean water and sanitation is a ubiquitous problem occurring globally. Addressing future challenges will require a combination of new technological development in water purification and environmental remediation technology with suitable conservation policies. In this scenario, new bioinspired materials will play a pivotal role in the development of more efficient and environmentally friendly solutions. The role of amphiphilic self-assembly on the fabrication of new biomimetic membranes for membrane separation like reverse osmosis is emphasized. Mesoporous support materials for semiconductor growth in the photocatalytic degradation of pollutants and new carriers for immobilization of bacteria in bioreactors are used in the removal and processing of different kind of water pollutants like heavy metals. Obstacles to improve and optimize the fabrication as well as a better understanding of their performance in small-scale and pilot purification systems need to be addressed. However, it is expected that these new biomimetic materials will find their way into the current water purification technologies to improve their purification/removal performance in a cost-effective and environmentally friendly way.

  19. [Immobilized microorganisms and water purification]. (United States)

    Mogilevich, N F


    Advantages and disadvantages of cells of aerobic microorganisms immobilized by the type of adhesion and incorporation into the gel beads, the amount of retained biomass, limitations of diffusion of oxygen and nutrients, viability, morphology, biochemical properties are described. Immobilized biocatalysts are discussed in the aspect of their use in purification of sewage waters.

  20. Purification and concentration of alphavirus. (United States)

    Lundstrom, Kenneth


    The alphaviruses Semliki Forest virus and Sindbis virus have been used frequently as expression vectors in vitro and in vivo. Usually, these systems consist of replication-deficient vectors that require a helper vector for packaging of recombinant particles. Replication-proficient vectors have also been engineered. Alphaviral vectors can be used as nucleic-acid-based vectors (DNA and RNA) or infectious particles. High-titer viral production is achieved in alphaviruses facilitates studies in mammalian and nonmammalian cell lines, primary cells in culture, and in vivo. The strong preference for expression in neuronal cells has made alphaviruses particularly useful in neurobiological studies. Unfortunately, their strong cytotoxic effect on host cells, relatively short-term transient expression patterns, and the reasonably high cost of viral production remain drawbacks. However, novel mutant alphaviruses have showed reduced cytotoxicity and prolonged expression. Membrane proteins (which are generally difficult to express at high levels in recombinant systems) have generated high yields and facilitate applications in structural biology. Alphaviruses have also been applied in vaccine development and gene therapy. Generally, purification or concentration of alphaviruses is not necessary. However, for instance, the medium derived from baby hamster kidney cells is toxic to primary neurons in culture. Including a purification step substantially improves the survival of the transduced neurons. Viral concentration and purification may also be advantageous for in vivo studies in animal models and are mandatory for clinical applications. This protocol describes three methods for purification and concentration of alphavirus.

  1. Biobased monoliths for adenovirus purification. (United States)

    Fernandes, Cláudia S M; Gonçalves, Bianca; Sousa, Margarida; Martins, Duarte L; Barroso, Telma; Pina, Ana Sofia; Peixoto, Cristina; Aguiar-Ricardo, Ana; Roque, A Cecília A


    Adenoviruses are important platforms for vaccine development and vectors for gene therapy, increasing the demand for high titers of purified viral preparations. Monoliths are macroporous supports regarded as ideal for the purification of macromolecular complexes, including viral particles. Although common monoliths are based on synthetic polymers as methacrylates, we explored the potential of biopolymers processed by clean technologies to produce monoliths for adenovirus purification. Such an approach enables the development of disposable and biodegradable matrices for bioprocessing. A total of 20 monoliths were produced from different biopolymers (chitosan, agarose, and dextran), employing two distinct temperatures during the freezing process (-20 °C and -80 °C). The morphological and physical properties of the structures were thoroughly characterized. The monoliths presenting higher robustness and permeability rates were further analyzed for the nonspecific binding of Adenovirus serotype 5 (Ad5) preparations. The matrices presenting lower nonspecific Ad5 binding were further functionalized with quaternary amine anion-exchange ligand glycidyltrimethylammonium chloride hydrochloride by two distinct methods, and their performance toward Ad5 purification was assessed. The monolith composed of chitosan and poly(vinyl) alcohol (50:50) prepared at -80 °C allowed 100% recovery of Ad5 particles bound to the support. This is the first report of the successful purification of adenovirus using monoliths obtained from biopolymers processed by clean technologies.

  2. A non-isotopic assay for histone deacetylase activity. (United States)

    Hoffmann, K; Brosch, G; Loidl, P; Jung, M


    Inhibitors of histone deacetylase (HD) bear great potential as new drugs due to their ability to modulate transcription and to induce apoptosis or differentiation in cancer cells. To study the activity of HD and the effect of potential inhibitors in vitro so far only radio-active assays have existed. For the search of new inhibitors and for the use in HD identification and purification we established a simple, non-radioactive assay that allows screening of large numbers of compounds. The assay is based on an aminocoumarin derivative of an Omega-acetylated lysine as enzyme substrate.

  3. Separation process design for isolation and purification of natural products

    DEFF Research Database (Denmark)

    Malwade, Chandrakant R.

    selection of separation techniques and operating conditions. The key factor in designing separation processes with multiple unit operations is to determine the synergy between them which in turn demands molecular level understanding of process streams. Therefore, the methodology is fortified with process......, thereby providing process information crucial for determining synergistic effects between different unit operations. In this work, the formulated methodology has been used to isolate and purify artemisinin, an antimalarial drug, from dried leaves of the plant Artemisia annua. A process flow sheet...... is generated consisting of maceration, flash column chromatography and crystallization unit operations for extraction, partial purification and final purification of artemisinin, respectively. PAT framework is used extensively to characterize the process streams at molecular level and the generated process...

  4. [Diversity and in vitro antitumnor activity of endophytic fungi from mangrove plants Xylocarpus]. (United States)

    Li, Ning; Ruan, Fei-Ying; Wen, Zheng-Shun; Li, Jian-Hua; Chen, Ri-Dao; Liu, Xiao; Xie, Dan; Li, Min-Yi; Wang, Chun-Mei; Wu, Jun; Dai, Jun-Gui


    A total of 24 biologically pure entophytic fungal strains were isolated from stems, leaves, and seed coats of Xylocarpus plants by repeated purification, and identified with Internal Transcribed Spacer (ITS) rDNA molecular method, which belonging to 14 genera, 11 families, 9 orders and 3 classes. There were differences in genus and species levels among three plant materials from different habitats and species, and it was found that the strains of Phomopsis and Colletotrichum existed in all three plant materials. In vitro assay of antitumor activity by MTT method revealed that the EtOAc extracts of 15 strains exhibited potent antitumor activity. These results suggest that it is of value for further investigation on the above fungal strains.

  5. Bioremediation of diesel-polluted soil using biostimulation as post-treatment after oxidation with Fenton-like reagents: assays in a pilot plant. (United States)

    Silva-Castro, Gloria Andrea; Rodelas, Belén; Perucha, Carlos; Laguna, Jaime; González-López, Jesús; Calvo, Concepción


    The present study focuses on the remediation of diesel-polluted soil using modified Fenton treatment coupled with inorganic NPK fertilizer ("Fenton+NPK"). Studies were carried out in a pilot plant containing 1 m(3) of sandy soil contaminated with 20,000 mg kg(-1) of diesel, placed outdoors at a temperature ranging between 5 and 10 °C. Results showed that NPK-fertilizer as post-treatment stimulated culturable degrading bacteria and enhanced dehydrogenase activity. Fenton+NPK treatment increased total petroleum hydrocarbon (TPH) removal efficacy. Natural attenuation removed 49% of TPH in the surface layer, 23% of TPH in the non-saturated layer and 4% of the TPH in the saturated layer, while the percentage removed of TPH after Fenton+NPK treatment was 58%, 57% and 32% respectively. The results from our study showed that, immediately after soil contamination, occurred a specialization and differentiation of the bacterial community, but after this initial modification, no significant changes of bacterial diversity was observed under natural attenuation conditions. In contrast, when the Fenton's reagent was applied a reduction of the bacterial biodiversity was observed. However, the post-biostimulation did enhance the degrading microbiota and stimulated their degrading biological activity. In conclusion, biostimulation, as a post-treatment step in chemical oxidation, is an effective solution to remediate hydrocarbon-polluted sites.

  6. Large-scale functional purification of recombinant HIV-1 capsid.

    Directory of Open Access Journals (Sweden)

    Magdeleine Hung

    Full Text Available During human immunodeficiency virus type-1 (HIV-1 virion maturation, capsid proteins undergo a major rearrangement to form a conical core that protects the viral nucleoprotein complexes. Mutations in the capsid sequence that alter the stability of the capsid core are deleterious to viral infectivity and replication. Recently, capsid assembly has become an attractive target for the development of a new generation of anti-retroviral agents. Drug screening efforts and subsequent structural and mechanistic studies require gram quantities of active, homogeneous and pure protein. Conventional means of laboratory purification of Escherichia coli expressed recombinant capsid protein rely on column chromatography steps that are not amenable to large-scale production. Here we present a function-based purification of wild-type and quadruple mutant capsid proteins, which relies on the inherent propensity of capsid protein to polymerize and depolymerize. This method does not require the packing of sizable chromatography columns and can generate double-digit gram quantities of functionally and biochemically well-behaved proteins with greater than 98% purity. We have used the purified capsid protein to characterize two known assembly inhibitors in our in-house developed polymerization assay and to measure their binding affinities. Our capsid purification procedure provides a robust method for purifying large quantities of a key protein in the HIV-1 life cycle, facilitating identification of the next generation anti-HIV agents.

  7. Development of RAP Tag, a Novel Tagging System for Protein Detection and Purification. (United States)

    Fujii, Yuki; Kaneko, Mika K; Ogasawara, Satoshi; Yamada, Shinji; Yanaka, Miyuki; Nakamura, Takuro; Saidoh, Noriko; Yoshida, Kanae; Honma, Ryusuke; Kato, Yukinari


    Affinity tag systems, possessing high affinity and specificity, are useful for protein detection and purification. The most suitable tag for a particular purpose should be selected from many available affinity tag systems. In this study, we developed a novel affinity tag called the "RAP tag" system, which comprises a mouse antirat podoplanin monoclonal antibody (clone PMab-2) and the RAP tag (DMVNPGLEDRIE). This system is useful not only for protein detection in Western blotting, flow cytometry, and sandwich enzyme-linked immunosorbent assay, but also for protein purification.

  8. [Efficiency of the preparation "Ekolan-M" for purification of oil polluted soil]. (United States)

    Nogina, T M; Dumanskaia, T U; Khomenko, L A; Podgorskiĭ, V S


    The efficiency of purification of oil contaminated loamy chernozem by the preparation "Ekolan-M" was investigated. During 12 months a complex soil bioremediation using the preparation and alfalfa, as the land-improving plant, at the final stage of purification resulted in the reduction of hydrocarbon content by 97.0%, and without the preparation - by 65.5 %. In the version of experiment with the preparation a 100% decrease of soil phytotoxicity was achieved and a significant stimulation of plant growth and development was observed. The process of soil purification was accompanied by intensive development of hydrocarbon-oxidizing microorganisms, the amount of which during the process of oil concentration gradually decreased, approaching the level in the control uncontaminated soil.

  9. Purification of an active fragment of Cry1Ie toxin from Bacillus thuringiensis. (United States)

    Guo, Shuyuan; Zhang, Chunlu; Lin, Xiaoyin; Zhang, Yanrui; He, Kanglai; Song, Fuping; Zhang, Jie


    The cry1I genes from Bacillus thuringiensis are a class of special genes with unique characteristics; they are silent in B. thuringiensis strains but can be over-expressed in Escherichia coli, resulting in a Cry1I-type protein with a molecular mass of approximately 81kDa. Cry1I-type protein is toxic to Lepidoptera larvae. A truncated Cry1Ie protein, IE648, which corresponds to the first 648 amino acids from the N-terminus of Cry1Ie, was purified from E. coli using Ni-NTA affinity isolation, Q-Sepharose Fast Flow chromatography, and Superdex-200 size-exclusion chromatography. It was determined using laboratory bioassays that the purified IE648 protein has good insecticidal activity. Heterologous competitive binding assays show that IE648 does not compete with Cry1Ac for binding to the brush border membrane vesicles of the Asian corn borer and does not compete with Cry1Ac at concentrations below a 500-fold excess of unlabeled Cry1Ac for binding to the peritrophic matrix of the insect. This result implies that IE648 may be a good candidate as part of a multiple-toxin strategy for the potential control of resistance in insect pests. The method of purification reported here is valuable for further research on the structure and function of IE648 and in evaluating the biosafety of this protein within transgenic plants. Copyright © 2011 Elsevier Inc. All rights reserved.

  10. Membrane adsorbers as purification tools for monoclonal antibody purification. (United States)

    Boi, Cristiana


    Downstream purification processes for monoclonal antibody production typically involve multiple steps; some of them are conventionally performed by bead-based column chromatography. Affinity chromatography with Protein A is the most selective method for protein purification and is conventionally used for the initial capturing step to facilitate rapid volume reduction as well as separation of the antibody. However, conventional affinity chromatography has some limitations that are inherent with the method, it exhibits slow intraparticle diffusion and high pressure drop within the column. Membrane-based separation processes can be used in order to overcome these mass transfer limitations. The ligand is immobilized in the membrane pores and the convective flow brings the solute molecules very close to the ligand and hence minimizes the diffusional limitations associated with the beads. Nonetheless, the adoption of this technology has been slow because membrane chromatography has been limited by a lower binding capacity than that of conventional columns, even though the high flux advantages provided by membrane adsorbers would lead to higher productivity. This review considers the use of membrane adsorbers as an alternative technology for capture and polishing steps for the purification of monoclonal antibodies. Promising industrial applications as well as new trends in research will be addressed.

  11. Study on the levels of activity of radionuclides in products solids of catalan water purification; Estudio sobre los niveles de actividad de radionuclidos en productos solidos de plants de potabilizacion de aguas catalanas

    Energy Technology Data Exchange (ETDEWEB)

    Montana, M.; Camacho, A.; Cespedes, R.; Devesa, R.; Serrano, I.; Duch, M. A.; Valles, I.


    In this work the results of radioactivity are presented in the sludge obtained part of the treatment process of 2 water treatment plants in Catalonia in which it is water from the rivers Ter and Llobregat. He has been also assessed the radiological impact of the sludge generated in these plants when used as raw material for the production of materials for the construction. (Author)

  12. Purification of Tetrahymena cytoskeletal proteins. (United States)

    Honts, Jerry E


    Like all eukaryotic cells, Tetrahymena thermophila contains a rich array of cytoskeletal proteins, some familiar and some novel. A detailed analysis of the structure, function, and interactions of these proteins requires procedures for purifying the individual protein components. Procedures for the purification of actin and tubulin from Tetrahymena are reviewed, followed by a description of a procedure that yields proteins from the epiplasmic layer and associated structures, including the tetrins. Finally, the challenges and opportunities for future advances are assessed.

  13. The recovery of cytokinins during extraction and purification of clubroot tissue

    NARCIS (Netherlands)

    Dekhuijzen, H.M.; Gevers, E.C.T.


    Losses of one naturally occurring cytokinin (zeatin) and one synthetic cytoknin (kinetin) were determined during purification of turnips (Brassica compestris) infected by Plasmodiophora brassicae (clubroot). A known amount of zeatin and 8‐14C‐kinetin was added after homogenization of plant material

  14. The recovery of cytokinins during extraction and purification of clubroot tissue

    NARCIS (Netherlands)

    Dekhuijzen, H.M.; Gevers, E.C.T.


    Losses of one naturally occurring cytokinin (zeatin) and one synthetic cytoknin (kinetin) were determined during purification of turnips (Brassica compestris) infected by Plasmodiophora brassicae (clubroot). A known amount of zeatin and 8‐14C‐kinetin was added after homogenization of plant material

  15. Comparing Russian and Finnish standards of water purification


    Maria, Pupkova


    The subject of this thesis is water purification. The first aim of this thesis is to consider different ways of water purification. The second aim is to compare Finnish and Russian standards of water purification. The third one is to show water purification methods on the pattern of Mikkeli water purification plan. Water purification methods of water intended for human consumption will be described.Combined tables will be done according to the quality requirement of drinking water of both,...

  16. Purification technology of molten aluminum

    Institute of Scientific and Technical Information of China (English)

    孙宝德; 丁文江; 疏达; 周尧和


    Various purification methods were explored to eliminate the dissolved hydrogen and nonmetallic inclusions from molten aluminum alloys. A novel rotating impeller head with self-oscillation nozzles or an electromagnetic valve in the gas circuit was used to produce pulse gas currents for the rotary impeller degassing method. Water simulation results show that the size of gas bubbles can be decreased by 10%-20% as compared with the constant gas current mode. By coating ceramic filters or particles with active flux or enamels, composite filters were used to filter the scrap A356 alloy and pure aluminum. Experimental results demonstrate that better filtration efficiency and operation performance can be obtained. Based on numerical calculations, the separation efficiency of inclusions by high frequency magnetic field can be significantly improved by using a hollow cylinder-like separator or utilizing the effects of secondary flow of the melt in a square separator. A multi-stage and multi-media purification platform based on these methods was designed and applied in on-line processing of molten aluminum alloys. Mechanical properties of the processed scrap A356 alloy are greatly improved by the composite purification.

  17. Technological assumptions for biogas purification. (United States)

    Makareviciene, Violeta; Sendzikiene, Egle


    Biogas can be used in the engines of transport vehicles and blended into natural gas networks, but it also requires the removal of carbon dioxide, hydrogen sulphide, and moisture. Biogas purification process flow diagrams have been developed for a process enabling the use of a dolomite suspension, as well as for solutions obtained by the filtration of the suspension, to obtain biogas free of hydrogen sulphide and with a carbon dioxide content that does not exceed 2%. The cost of biogas purification was evaluated on the basis of data on biogas production capacity and biogas production cost obtained from local water treatment facilities. It has been found that, with the use of dolomite suspension, the cost of biogas purification is approximately six times lower than that in the case of using a chemical sorbent such as monoethanolamine. The results showed travelling costs using biogas purified by dolomite suspension are nearly 1.5 time lower than travelling costs using gasoline and slightly lower than travelling costs using mineral diesel fuel.

  18. Sewage Purification Business Process Management

    Directory of Open Access Journals (Sweden)

    Esad Ahmetagić


    Full Text Available This paper presents the current level of drainage and sewage purification facilities built in the Autonomous Province of Vojvodina, a territorial unit of the Republic of Serbia. It also points out the issues related to organized business management in companies involved in this business.The management of business processes in sewage purification involves a comprehensive cycle: business organizing process, issues of standard, investments, workforce, and information system design as factors in establishing an effective organization of business processes. The definition of gap existing between the current approach to organizing business activities and the need to establish an approach based on knowledge, information technologies, and effective business process management points to the necessity for organization redesign and standard definition in business process management. Sewage purification business process management in Vojvodina, the Republic of Serbia has been elaborated through theoretical presentation and a practical example realized by electronic ISO 9001:2008 system of quality management in public water utility company JKP "Vodokanal" Sombor.

  19. Nanostructured Catalytic Reactors for Air Purification Project (United States)

    National Aeronautics and Space Administration — This SBIR Phase I project proposes the development of lightweight compact nanostructured catalytic reactors for air purification from toxic gaseous organic...

  20. Nanostructured Catalytic Reactors for Air Purification Project (United States)

    National Aeronautics and Space Administration — This SBIR Phase II project proposes the development of lightweight compact nanostructured catalytic reactors for air purification from toxic gaseous organic...

  1. Characterization and purification of bacteriophages using chromatofocusing. (United States)

    Brorson, Kurt; Shen, Hong; Lute, Scott; Pérez, Jessica Soto; Frey, Douglas D


    The technique of chromatofocusing was applied to the characterization and purification of three bacteriophages that are routinely used for testing virus filters: phiX174, PR772, and PP7. Chemically well-defined eluent buffers were used, instead of the more commonly used chromatofocusing polyampholyte buffers. Chromatographic column packings were selected to minimize band broadening by confining bacteriophage adsorption solely to the exterior particle surface. Under the conditions used it was determined that bacteriophages could be made to focus into narrow bands in a retained pH gradient with recoveries of live phage that ranged from 15 to nearly 100% as determined by a plaque-forming assay. Retention times and apparent isoelectric point data were obtained for samples consisting either of purified bacteriophage, or samples consisting of crude preparations of bacteriophages containing host cell impurities. Isoelectric point estimates were obtained using modified, previously described models. The results obtained suggest that chromatofocusing is a simple and rapid method for obtaining approximate isoelectric points for bacteriophages and probably other types of viruses. It is also likely a useful method for purifying these materials.

  2. Purification and properties of thioether methyltransferase

    Energy Technology Data Exchange (ETDEWEB)

    Mozier, N.M.


    A method to assay activity was developed which measures acceptance of methyl groups from (methyl-{sup 3}H)-S-adenosylmethionine by dimethyl selenide. The product, ({sup 3}H)trimethylselenonium ion, is separated by HPLC and quantitated by scintillation counting. Thioether methyltransferase from mouse liver and lung resides primarily in the cytosol. In terms of specific activity the enzyme is most active in the lung and liver. Purification from lung cytosol requires a three-step process of DEAE and gel filtration column chromatographies followed by chromatofocusing. SDS-Polyacrylamide gel electrophoresis shows a single homogeneous band with a molecular mass of 28,000 daltons. Vmax and Km values for dimethyl selenide as a substrate are 15. 7 pmol/min and 0.44 {mu}M, respectively. Our studies have also shown that this purified enzyme is capable of methylating a wide range of compounds. To further test the enzyme's role in detoxification, in vivo studies were performed by injecting mice with substrate and (methyl-{sup 3}H)methionine and analyzing tissue extracts and urine for (methyl-{sup 3}H)sulfonium.

  3. A reversible Renilla luciferase protein complementation assay for rapid identification of protein-protein interactions reveals the existence of an interaction network involved in xyloglucan biosynthesis in the plant Golgi apparatus. (United States)

    Lund, Christian H; Bromley, Jennifer R; Stenbæk, Anne; Rasmussen, Randi E; Scheller, Henrik V; Sakuragi, Yumiko


    A growing body of evidence suggests that protein-protein interactions (PPIs) occur amongst glycosyltransferases (GTs) required for plant glycan biosynthesis (e.g. cell wall polysaccharides and N-glycans) in the Golgi apparatus, and may control the functions of these enzymes. However, identification of PPIs in the endomembrane system in a relatively fast and simple fashion is technically challenging, hampering the progress in understanding the functional coordination of the enzymes in Golgi glycan biosynthesis. To solve the challenges, we adapted and streamlined a reversible Renilla luciferase protein complementation assay (Rluc-PCA), originally reported for use in human cells, for transient expression in Nicotiana benthamiana. We tested Rluc-PCA and successfully identified luminescence complementation amongst Golgi-localizing GTs known to form a heterodimer (GAUT1 and GAUT7) and those which homooligomerize (ARAD1). In contrast, no interaction was shown between negative controls (e.g. GAUT7, ARAD1, IRX9). Rluc-PCA was used to investigate PPIs amongst Golgi-localizing GTs involved in biosynthesis of hemicelluloses. Although no PPI was identified among six GTs involved in xylan biosynthesis, Rluc-PCA confirmed three previously proposed interactions and identified seven novel PPIs amongst GTs involved in xyloglucan biosynthesis. Notably, three of the novel PPIs were confirmed by a yeast-based split-ubiquitin assay. Finally, Gateway-enabled expression vectors were generated, allowing rapid construction of fusion proteins to the Rluc reporters and epitope tags. Our results show that Rluc-PCA coupled with transient expression in N. benthamiana is a fast and versatile method suitable for analysis of PPIs between Golgi resident proteins in an easy and mid-throughput fashion in planta.

  4. Breathing Air Purification for Hyperbaric Purposes, Part II

    Directory of Open Access Journals (Sweden)

    Woźniak Arkadiusz


    Full Text Available Determining the efficiency of breathing air purification for hyperbaric purposes with the use of filtration systems is of a crucial importance. However, when the Polish Navy took samples of breathing air from their own filtration plant for quality purposes, these were found to not meet the required standard. The identification of this problem imposed the need to undertake actions aimed at the elimination of the identified disruptions in the process of breathing air production, with the objective of assuring its proper quality. This study presents the results of the initial tests on the air supply sources utilised by the Polish Navy, which were carried out for the purpose of setting a proper direction of future works and implementing corrective measures in order to optimise the breathing air production process. The obtained test results will be used in a subsequent publication devoted to the assessment of the level of efficiency of air purification with the use of a multifaceted approach consisting in the utilisation of various types of air supply sources and different configurations of purification systems.

  5. Production and purification of polyclonal antibody against melatonin hormone

    Directory of Open Access Journals (Sweden)

    Fooladsaz K


    Full Text Available Nowadays immunochemical techniques have played a very important and valuable role in quantitative and qualitative assays of liquid compounds of the body. Producing antibody against immunogenes is the first step to make immunochemical kits. In this study production and purification of polyclonal antibody against melatonin has been considered. This hormone which has several important functions in physiological conditions such as migraine, cirrhosis, mammary gland cancer and other diseases, is the most important pineal gland secretion. This gland is a circumventricular organ of brain and according to histological and anatomical studies, it is a high secretory organ, that secretes active biological substances like melatonin, oxytocin, serotonin and ect. In this study, melatonin has been considered as hapten and has become an immunogen by being linked to the bovine serum Albumin. Then, by the immunization of three white New Zeland rabbits that had the booster injections in regular intervals, the antibody titer was detected to be 1/2000, by using checkboard curves, and with the use of melatonin linked to penicillinase as a labeled antigen, the titer was detected 1/200. Finally an antibody with high purification rate has been obtained, which can be used in immunochemical assays like RIA, ELISA, and EIA.

  6. Recovery of radiogenic lead-208 from a residue of thorium and rare earths obtained during the operation of a thorium purification pilot plant; Separacao e recuperacao de chumbo-208 dos residuos de torio terras raras gerados na unidade piloto de purificacao de nitrato de torio

    Energy Technology Data Exchange (ETDEWEB)

    Seneda, Jose Antonio


    Brazil has a long tradition in thorium technology, from mineral dressing (monazite) to the nuclear grade thorium compounds. The estimate reserves are 1200,000. ton of ThO{sub 2}. As a consequence from the work of thorium purification pilot plant at Instituto de Pesquisas Energeticas e Nucleares-CNEN/IPEN-SP, about 25 ton of a sludge containing thorium and rare earths was accumulated. It comes as a raffinate and washing solutions from thorium solvent extraction. This sludge, a crude hydroxide named RETOTER contains thorium, rare earths and minor impurities including the radiogenic lead-208, with abundance 88.34 %. This work discusses the results of the studies and main parameters for its recovery by anionic ion exchange technique in the hydrochloric system. The isotope abundance of this lead was analyzed by high resolution mass spectrometer (ICPMS) and thermoionic mass spectrometer (TIMS) and the data was used to calculate the thermal neutron capture cross section. The value of {sigma}{gamma}{sup 0} = 14.6{+-}0.7 mb was found, quite different from the {sigma}{gamma}{sup 0} = 174.2 {+-} 7.0 mb measure cross section for the natural lead. Preliminary study for the thorium and rare earths separation and recovery was discussed as well. (author)

  7. Function and anatomy of plant siRNA pools derived from hairpin transgenes

    Directory of Open Access Journals (Sweden)

    Lee Kevin AW


    Full Text Available Abstract Background RNA interference results in specific gene silencing by small-interfering RNAs (siRNAs. Synthetic siRNAs provide a powerful tool for manipulating gene expression but high cost suggests that novel siRNA production methods are desirable. Strong evolutionary conservation of siRNA structure suggested that siRNAs will retain cross-species function and that transgenic plants expressing heterologous siRNAs might serve as useful siRNA bioreactors. Here we report a detailed evaluation of the above proposition and present evidence regarding structural features of siRNAs extracted from plants. Results Testing the gene silencing capacity of plant-derived siRNAs in mammalian cells proved to be very challenging and required partial siRNA purification and design of a highly sensitive assay. Using the above assay we found that plant-derived siRNAs are ineffective for gene silencing in mammalian cells. Plant-derived siRNAs are almost exclusively double-stranded and most likely comprise a mixture of bona fide siRNAs and aberrant partially complementary duplexes. We also provide indirect evidence that plant-derived siRNAs may contain a hitherto undetected physiological modification, distinct from 3' terminal 2-O-methylation. Conclusion siRNAs produced from plant hairpin transgenes and extracted from plants are ineffective for gene silencing in mammalian cells. Thus our findings establish that a previous claim that transgenic plants offer a cost-effective, scalable and sustainable source of siRNAs is unwarranted. Our results also indicate that the presence of aberrant siRNA duplexes and possibly a plant-specific siRNA modification, compromises the gene silencing capacity of plant-derived siRNAs in mammalian cells.

  8. Hemolytic plate assay for quantification of active human complement component C3 using methylamine-treated plasma as complement source

    DEFF Research Database (Denmark)

    Ploug, M; Jessen, T E; Welinder, K. G.


    A hemolytic plate assay specific for active human complement component C3 is described. The method is well suited for tracing active C3 during preparative purification or for screening of plasma samples. The assay is based on activation of the alternative pathway of complement by unmodified rabbi...

  9. 烟气CO2捕集胺液净化回用技术%Amine purification technology for CO2 capture system in coal-fired power plants

    Institute of Scientific and Technical Information of China (English)

    张小刚; 张一; 毛小雨; 李建玺; 李爽佩; 张安琪; 何涌; 张弛


    针对CO2捕集系统有机胺液中热稳定盐的积累进而引起胺液损耗增加、腐蚀严重等问题,提出采用阴离子交换树脂去除胺液中热稳定盐阴离子,采用活性炭吸附去除胺液色度,最后采用大孔树脂进一步净化胺液的组合工艺,并对工艺的影响因素,以及净化胺液的吸收性能进行了测试。结果表明:阴离子交换树脂处理工艺在较高的温度、pH 值及含盐量水平的条件下,均可以达到较好的效果,热稳盐去除率可达90%以上;变质有机胺液经过处理后,其 CO2吸附能力恢复至新鲜胺液的92%,并且溶液颜色恢复至淡黄色透明液体,可满足工业回用要求。%The accumulation of heat stable salts (HSS)in amine liquid during CO2 capture cycle can increase amine liquid consumption and cause serious corrosion.Thus,a combined process was developed,including removal of HSS anion by anion exchange resin,chromaticity color by activated carbon,and further purifica-tion by macro-porous resin.Some influencing factors were investigated and the absorption performance of purified amine was tested.The results show that,the amine exchange resin can perform well even under conditions with high temperature,high pH value and high salt content,of which the removal rate of HSS was over 90%.After treated by the combined process above,the CO2 absorption ability of the purified a-mine liquid can recover to 9 2% of that of the flesh amine liquid,with a color of transparent pale yellow, which meets the requirements of industrial recycling.

  10. Abundances of tetracycline, sulphonamide and beta-lactam antibiotic resistance genes in conventional wastewater treatment plants (WWTPs) with different waste load

    DEFF Research Database (Denmark)

    Laht, Mailis; Karkman, Antti; Voolaid, Veiko


    Antibiotics and antibiotic resistant bacteria enter wastewater treatment plants (WWTPs), an environment where resistance genes can potentially spread and exchange between microbes. Several antibiotic resistance genes (ARGs) were quantified using qPCR in three WWTPs of decreasing capacity located...... conclude that there is neither considerable enrichment nor purification of antibiotic resistance genes in studied conventional WWTPs....... in Helsinki, Tallinn, and Tartu, respectively: sulphonamide resistance genes (sul1 and sul2), tetracycline resistance genes (tetM and tetC), and resistance genes for extended spectrum beta-lactams (blaoxa-58, blashv-34, and blactx-m-32). To avoid inconsistencies among qPCR assays we normalised the ARG...

  11. Plant Litter Submergence Affects the Water Quality of a Constructed Wetland

    National Research Council Canada - National Science Library

    Pan, Xu; Ping, Yunmei; Cui, Lijuan; Li, Wei; Zhang, Xiaodong; Zhou, Jian; Yu, Fei-Hai; Prinzing, Andreas


      Plant litter is an indispensable component of constructed wetlands, but how the submergence of plant litter affects their ecosystem functions and services, such as water purification, is still unclear...

  12. Biogas Purification up to Final Product

    Directory of Open Access Journals (Sweden)

    Yu. Losiuk


    Full Text Available The paper considers main technological methods for biogas purification from impurities that permit to increase energy value of the product and decrease its corrosion activity.  While evaluating economic efficiency due to introduction of the corresponding purification technology, in addition, it is necessary to take into account an ecological factor.

  13. Purification and characterization of a thermostable glucoamylase ...

    African Journals Online (AJOL)



    Jun 7, 2010 ... tryptophan and serine residues in the catalytic process. Raw corn .... selected for amylase production, purification and characterization. .... chromatogram was developed with solvent system of butanol/ ... starch affinity and acetone precipitation method for ..... Optimization of extraction and purification of.

  14. Boron carbide morphology changing under purification (United States)

    Rahmatullin, I. A.; Sivkov, A. A.


    Boron carbide synthesized by using coaxial magnetoplasma accelerator with graphite electrodes was purified by two different ways. XRD-investigations showed content changing and respectively powder purification. Moreover TEM-investigations demonstrated morphology changing of product under purification that was discussed in the work.


    DEFF Research Database (Denmark)


    Purification of fluoride contaminated magadi is studied using bone char sorption and calcium precipitation. The bone char treatment is found to be workable both in columns and in batches where the magadi is dissolved in water prior to treatment. The concentrations in the solutions were 89 g magadi...... treatment method. A procedure for purification of fluoride contaminated magadi at household level is described....

  16. Partial Purification and Characterization of Extracellular Protease ...

    African Journals Online (AJOL)


    Purification of the enzyme by gel filtration chromatography on Sephadex G75 following ammonium sulphate precipitation gave 2.26 fold increase in purification with specific activity of 46.13 .... minutes in a water bath and were allowed to cool.

  17. Cloning, purification, crystallization and 1.57 Å resolution X-ray data analysis of AmsI, the tyrosine phosphatase controlling amylovoran biosynthesis in the plant pathogen Erwinia amylovora. (United States)

    Benini, Stefano; Caputi, Lorenzo; Cianci, Michele


    The Gram-negative bacterium Erwinia amylovora is a destructive pathogen of plants belonging to the Rosaceae family. Amongst its pathogenicity factors, E. amylovora produces the exopolysaccharide amylovoran, which contributes to the occlusion of plant vessels, causing wilting of shoots and eventually resulting in plant death. Amylovoran biosynthesis requires the presence of 12 genes (from amsA to amsL) clustered in the ams region of the E. amylovora genome. They mostly encode glycosyl transferases (AmsG, AmsB, AmsD, AmsE, AmsJ and AmsK), proteins involved in amylovoran translocation and assembly (AmsH, AmsL and AmsC), and also a tyrosine kinase (AmsA) and a tyrosine phosphatase (AmsI), which are both involved in the regulation of amylovoran biosynthesis. The low-molecular-weight protein tyrosine phosphatase AmsI was overexpressed as a His6-tagged protein in Escherichia coli, purified and crystallized. X-ray diffraction data were collected to a maximum resolution of 1.57 Å in space group P3121.

  18. 超滤+反渗透在净水厂脱盐深度处理的应用%The Application of Ultrafiltration and Reverse Osmosis Treatment in the Depth Desalting Processing of Purification Plant

    Institute of Scientific and Technical Information of China (English)

    姚琦; 张继昌; 佘丽华


    In view of the actual situation that the water near the beach, has high salinity (chloride), Hangfeng plant project phaseⅡusing ultrafiltration and reverse osmosis treatment, ob-tains bet er effect of desalination, water treatment plant efflue-nt reached standards. This paper introduces the whole plant and ultrafiltration and reverse osmosis treating process flow, d-esign parameters and treatment effect, in order to more widely applied in the treatment project of high containing water.%针对水源靠近海边、含盐量(氯化物)较高的实际情况,航丰水厂二期工程采用超滤+反渗透深度处理工艺,取得较好脱盐效果,水厂出水稳定达标。文章介绍了整个水厂及超滤+反渗透深度处理工艺流程、设计参数及处理效果,以期更广泛地应用于高含盐水处理工程。

  19. 紫外交联合并串联亲和纯化高效鉴定蛋白复合物组分的RNA结合活性%Efficient Assay of The RNA-binding Activities in a Protein Complex by UV Cross-linking and Tandem Affinity Purification

    Institute of Scientific and Technical Information of China (English)

    岳莎; 周淑如; 郭学敏


    RNA结合蛋白在RNA的生成与代谢中发挥着重要作用.我们在近年报道的PAR-CLIP (photoactivatableribonucleoside-enhanced crosslinking and immunoprecipitation)技术的基础上建立了一套快速、有效鉴定RNA结合蛋白的实验方法:以串联亲和纯化替代一步免疫沉淀获得高纯度蛋白-RNA复合物;将Sypro Ruby蛋白染色与RNA放射自显影相结合判断复合物中哪种或哪些组分为RNA结合蛋白,该方法命名为紫外交联合并的串联亲和纯化(cross-linking and tandem affinity purification,CLiTAP).运用该方法对布氏锥虫的三种锌指蛋白ZC3H7、ZC3H34和ZC3H5进行分析,发现ZC3H7作为帽结合蛋白复合物的核心组分具有很强的RNA结合能力;ZC3H34结合RNA能力较弱,但其互作蛋白具有强的RNA结合活性;相比之下,ZC3H5及其复合物组分皆无RNA结合活性.这些结果表明,CLiTAP与蛋白质鉴定方法相结合,能够有效鉴定靶蛋白复合物中的RNA结合蛋白种类,也为进一步定位RNA结合位点、研究RNA结合蛋白的结构及作用机制奠定了基础.%RNA-binding proteins (RBPs) function importantly in RNA synthesis and metabolism by interacting with specific RNAs.The identification of RNA-protein interactions is required for understanding the cellular function mechanisms.A new technique,PAR-CLIP (photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation),has been used to determine the target transcripts of a few RBPs and map the RNA binding sites in a genome-wide fashion.In this study,a rapid and effective method was developed on the basis of PAR-CLIP to detect the RNA binding activity of putative RBPs and their complexes,termed UV cross-linking and tandem affinity purification (CLiTAP).The improvements include:(1) Tandem affinity purification was performed to purify RNA-protein complexes efficiently; (2) Sypro Ruby staining and autoradiography were used sequentially to determine which protein(s) possessing RNA

  20. High yield purification of full-length functional hERG K+ channels produced in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Molbaek, Karen; Scharff-Poulsen, Peter; Hélix-Nielsen, Claus;


    knowledge this is the first reported high-yield production and purification of full length, tetrameric and functional hERG. This significant breakthrough will be paramount in obtaining hERG crystal structures, and in establishment of new high-throughput hERG drug safety screening assays....

  1. 自然沟渠与水泥沟渠水生植物群落结构及净水效果研究%Study on the community structure of aquatic plant and the effect of water purification in natural ditch and cement ditch

    Institute of Scientific and Technical Information of China (English)

    刘丰雷; 谢从新; 张念; 吕元蛟; 张志敏; 吴强亮


    Through field investigating,the community structure and biomass of aquatic plant and the effect of Natural Ditch (ND) and Cement Ditch (CD) on water quality were studied to improve basis for building ecological ditch in the aquaculture farm.Studies showed:it comprised 17 families,23 genus,24 species,and consisted of Alternanthera philoxeroides-Zizania latifolia community,Alternanthera philoxeroide-Nelumbo nucifera community in ND,where the basal coverage of aquatic plant is more than 50% and the average of biomass is 2 633.85 g/m2 ; it comprised 7 families,7 genus,7 species,and composed of Valisneria batans community,Ceratophyllum demersum community in CD,where the basal coverage is less than 20% and the biodiversity is greater than CD.The concentration of TN in ND and CD are 0.97 ±0.32 mg/L,1.48 ±0.61 mg/L respectively; the concentration of TP is(0.13 ± 0.06)mg/L,(0.16 ± 0.07)mg/L respectively; the concentration of Chl-a is(35.12 ± 17.73) μg/L,(69.24 ± 17.31) μg/L respectively; the concentration of DO is (3.67 ± 2.17)mg/L,(6.12 ± 1.95)mg/L respectively.The aquatic plant is rich and the effect of water quality purification is obvious in ND,but DO is substantially lower than CD.Considering system stability,cost of building and effect of water quality purification,suggest to employ ND.%采用野外调查法,对养殖场自然沟渠与水泥沟渠的水生植物群落结构、生物量及其对水质的影响进行比较研究.结果表明:自然沟渠全年水生植物种类7科23属24种,以空心莲子草-菰群和空心莲子草-莲群为主,平均生物量为2 633.85 g/m2,真盖度>50%;水泥沟渠7科7属7种,以苦草群、金鱼藻群为主,真盖度<20%;自然沟渠生物多样性显著高于水泥沟渠.自然沟渠和水泥沟渠的TN分别为(0.97±0.32) mg/L和(1.48±0.61) mg/L,TP分别为(0.13±0.06) mg/L和(0.16±0.07)mg/L,Chl-a分别为(35.12±17.73)μg/L和(69.24±17.31) μg/L,DO分别为(3.67±2.17) mg/L和(6.12±1.95) mg/L

  2. TMI-2 purification demineralizer resin study

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, J D; Osterhoudt, T R


    Study of the Makeup and Purification System demineralizers at TMI-2 has established that fuel quantities in the vessels are low, precluding criticality, that the high radioactive cesium concentration on the demineralizer resins can be chemically removed, and that the demineralizer resins can probably be removed from the vessels by sluicing through existing plant piping. Radiation measurements from outside the demineralizers establishing that there is between 1.5 and 5.1 (probably 3.3) lb of fuel in the A vessel and less than that amount in the B vessel. Dose rates up to 2780 R per hour were measured on contact with the A demineralizer. Remote visual observation of the A demineralizer showed a crystalline crust overlaying amber-colored resins. The cesium activity in solid resin samples ranged from 220 to 16,900 Based on this information, researchers concluded that the resins cannot be removed through the normal pathway in their present condition. Studies do show that the resins will withstand chemical processing designed to rinse and elute cesium from the resins. The process developed should work on the TMI-2 resins.

  3. Purification of tomato yellow leaf curl geminivirus. (United States)

    Luisoni, E; Milne, R G; Vecchiati, M


    Attempts were made to find a good purification procedure for tomato yellow leaf curl virus (TYLCV), a dangerous and continuously spreading whitefly-transmitted germinivirus, up to now only partially purified. Electron microscopy, serology and spectrophotometry were used to evaluate different procedures. The scheme finally adopted was the following: collect leaves and stems from Nicotiana benthamiana graft-infected 45-60 days previously (5-10 g/plant); homogenize with 0.5 M phosphate buffer pH 6 containing 2.5 mM NaEDTA, 10 mM Na2SO3, 0.1% 2-mercaptoethanol, 1% Triton X-100 and 0.1% Driselase (3-4 ml of buffer for each g of material); incubate overnight on ice with gentle agitation; filter; emulsify with 15% cold chloroform; centrifuge at low speed; ultracentrifuge supernatant; resuspend pellets in 0.5 M phosphate buffer pH 7 containing 2.5 mM NaEDTA; centrifuge at low speed; repeat resuspension of the pellets and low-speed centrifugation; ultracentrifuge the pooled supernatant on a Cs2SO4 gradient (e.g. for 5 h at 41,000 rpm); collect the virus band and dialyse or ultracentrifuge the virus. The virus yield was 5-10 mg per kg of tissue.

  4. Seperation and Purification of Plant Insulin and Its Hypoglycemic Effect on the Experimontal Animals%植物胰岛素的分离纯化及降糖作用的实验动物研究

    Institute of Scientific and Technical Information of China (English)

    徐军; 吴鹏; 韩林; 张忠; 黄竹青


    目的:探寻大量纯化苦瓜中植物胰岛素的技术方法.方法:采用盐析、阴离子交换层析和凝胶过滤层析三步技术纯化苦瓜中植物胰岛素,并行电泳、层析和动物实验鉴定,壳聚糖包封.结果:目的提取物于11000处出现一条电泳带,相对迁移率与胰岛素相同;壳聚糖包封后苦瓜中植物胰岛素的降糖作用更显著.结论:本方法可适用于大量纯化苦瓜中植物胰岛素.%Objective: To explore a technical method to purify the plant insulin in the bitter melon. Methods: applying three - step methods to purify insulin from bitter melon these are salting out, ion exchange chromatography and gel filtration chromatography. And than using electrophoresis chromatography, animal experiments and chitosan envelopment methods to identify it. Results:The extracts appeared electrophoresis band at the 11000 molecular weight, and had the same relative mobility with insulin. The plant insulin from the bitter melons has the hypoglycemic effect significantly after the chitosan envelopment methods. Conclusion: To stabilized extract the plant insulin from the bitter melon can depend on this method mentiond in this paper.

  5. 绿萝等6种室内观赏植物对氨气净化作用分析%Analysis of A Purification Effect of Ammonia on Epipremnum Aureum and Other Indoor Ornamental Plants

    Institute of Scientific and Technical Information of China (English)

    崔洪珊; 贾伟


    选取绿萝(Epipremnum aureum)、金边虎皮兰(Sansevieria trifasciata Lanrentii)、常春藤(Hedera nepalensis var.Sinensis)、豹纹竹芋(Maranta leuconeura)、金边吊兰(Phnom Penh Chlorophytum)、芦荟(Aloe vera var.Chinensis)六种植物,采用熏蒸方式,研究它们在.6 h熏蒸时间内,对氨气的净化效果.实验结果表明,6种植物对氨气均有净化作用,但不同植物净化效果有显著性差异.吸附氨气能力较强的为绿萝、常春藤;.中等的为豹纹竹芋、金边虎皮兰;.较弱的为金边吊兰、芦荟.%The research selectedsix kinds of plantssuch as Epipremnum aureum, Sansevieria trifasciata Lanrentii, Hedera nepalensis var. Sinensis, Maranta leuconeura, Phnom Penh ChlorophytumandAloe vera var. Chinensis, and used fumigation to study the purifying effect to Ammonia of above plant within 6 h fumigation time. The results show that 6 kinds of plants all have purifying effect to Ammonia, but there are significant differences between different plants. The Epipremnum aureum and Hedera nepalensis var. Sinensis has strong adsorption ability to Ammonia, medium for Maranta leuconeuraand Sansevieria trifasciata Lanrentii, and weaker for Phnom Penh Chlorophytum and Aloe vera var. Chinensis.

  6. Purification of anaerobic digestion biogas from a wastewater treatment plant for its use as bio fuel; Purificacion del biogas de digestion anaerobia de una depuradora de aguas residuales para uso como biocombustible

    Energy Technology Data Exchange (ETDEWEB)

    Osorio Robles, F.; Torres Rojo, J. C.; Sanchez Bas, M.; Moya Sanchez, N.


    The first phase of the investigation whose results are presented in this article, consists on the optimization of the biogas desulphurization. In our case this process was made by chemical way. Besides the scrubbing towers, the pilot plant used included filters of activated carbon at the end of the line. The H{sub 2}S inflow concentrations were quite high. After the carried out rehearsals, the effluent biogas from the scrubbing towers presents a H{sub 2}S concentration less than 1 ppm and zero or undetectable values of up to fifty eight analyzed trace elements. (Author) 12 refs.

  7. 茶树酯型儿茶素水解酶鉴定及其检测体系的建立%Identification and Reaction Assay of Galloylated Catechins Hydrolase in Tea Plant

    Institute of Scientific and Technical Information of China (English)

    聂志银; 刘亚军; 刘莉; 高丽萍; 夏涛


    In this experiment, high activity of galloylated catechins hydrolase (GCH) was detected existing in tea plant (Camellia sinensis (L.) O. Kuntze) by enzymology analysis in vitro, combining thin layer chromatography (TLC), high performance liquid chromatography (HPLC), and liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis. The galloylated catechins could be hydrolyzed to ungalloylated catechins and gallic acid (GA) with the GCH action. The optimum reaction assay of GCH has been established. The 2.5 mL enzyme reaction assay included 0.1 mol/L phosphate buffer (pH 6.5), 0.2 mmol/L EGCG, 2.4 mmol/L sodium ascorbate, crude enzyme extract (0.1 mg total protein), and then it was incubated at 30 ℃ for 30 min. Besides, the crude enzyme extract was partially purified via ammonium sulfate fractionation, anion exchange chromatography on Q Sepharose Fast Flow column and gel filtration chromatography on superdex 200 sequentially.%本试验利用体外酶学方法,结合薄层层析( TLC)、高效液相色谱(HPLC)和液相色谱-串联质谱(LC-MS/MS)分析,首次从茶树中检测到活性较高的酯型儿茶素水解酶(Galloylated Catechins Hydrolase,GCH)的存在.在酯型儿茶素水解酶催化下,酯型儿茶素发生水解反应,生成没食子酸(GA)和非酯型儿茶素.试验确立了酯型儿茶素水解酶的最适检测体系,在2.5 mL反应体系中包含0.2 mmol/L酯型儿茶素、2.4 mmol/L抗坏血酸、粗酶提取液若干(含0.1 mg酶蛋白)和0.1 mol/L磷酸缓冲液(pH6.5),在30℃下,反应30 min.此外,试验利用硫酸铵分级沉淀、阴离子交换层析和凝胶过滤层析对该酶进行了初步纯化.

  8. Purification in a functional form of the terminal protein of Bacillus subtilis phage phi 29.



    Phage phi 29 terminal protein, p3, essentially pure, was isolated in a denatured form from viral particles, and anti-p3 antiserum was obtained. A radioimmunoassay to detect and quantitate protein p3 was developed. By using this assay, native protein p3 was highly purified from Escherichia coli cells harboring a gene 3-containing recombinant plasmid. After three purification steps, the protein was more than 96% pure; its amino acid composition was very similar to that deduced from the nucleoti...

  9. Purification in a functional form of the terminal protein of Bacillus subtilis phage ø29



    Phage phi 29 terminal protein, p3, essentially pure, was isolated in a denatured form from viral particles, and anti-p3 antiserum was obtained. A radioimmunoassay to detect and quantitate protein p3 was developed. By using this assay, native protein p3 was highly purified from Escherichia coli cells harboring a gene 3-containing recombinant plasmid. After three purification steps, the protein was more than 96% pure; its amino acid composition was very similar to that deduced from the nucleoti...

  10. Herbicide resistance screening assay. (United States)

    Peterson, Joan M


    Herbicide resistance screening is a method that can be used not only to determine presence of the enzyme, phosphinothricin acetyltransferase, encoded by either the Bar or the Pat gene in transgenic maize, but also to assess the inheritance ratio of those genes in a segregating population. Herbicide screening can also be used to study linkage of a transgene of interest that was cotransformed with the herbicide resistance marker gene. By combining the herbicide screen assay with a PCR-based screen of leaf tissue DNA for the presence of both the Bar or the Pat gene marker and a cotransformed transgene of interest from the same seedling tissue and maintaining that seedling identity, the researcher can identify linkage or the possible breakdown in linkage of the marker gene and the transgene of interest. Further, the occurrence of "DNA silencing" can be evaluated if an individual seedling that was susceptible to the applied herbicide nonetheless gave PCR data that indicated presence of the gene responsible for herbicide resistance. Similarly, "DNA silencing" of the gene of interest may be investigated if the seeds can be screened and scored for that phenotypic trait in a nondestructive manner prior to planting.

  11. Investigation of purification process stresses on erythropoietin peptide mapping profile. (United States)

    Sepahi, Mina; Kaghazian, Hooman; Hadadian, Shahin; Norouzian, Dariush


    Full compliance of recombinant protein peptide mapping chromatogram with the standard reference material, is one of the most basic quality control tests of biopharmaceuticals. Changing a single amino acid substitution or side chain diversity for a given peptide changes protein hydrophobicity and causes peak shape or retention time alteration in a peptide mapping assay. In this work, the effect of different stresses during the recombinant erythropoietin (EPO) purification process, including pH 4, pH 5, and room temperature were checked on product peptide mapping results. Cell culture harvest was purified under stress by different chromatographic techniques consisting of gel filtration, anionic ion exchange, concentration by ultrafiltration, and high resolution size exclusion chromatography. To induce more pH stresses, the purified EPO was exposed to pH stress 4 and 5 by exchanging buffer by a 10 KDa dialysis sac overnight. The effects of temperature and partial deglycosylation (acid hydrolysis) on purified EPO were also studied by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and peptide mapping analysis. Removal of sialic acid by mild hydrolysis was performed by exposure to two molar acetic acid at 80°C for 3 h. No significant effect was observed between intact and stressed erythropoietin peptide mapping profiles and SDS-PAGE results. To validate the sensibility of the technique, erythropoietin was partially acid hydrolyzed and significant changes in the chromatographic peptide map of the intact form and a reduction on its molecular weight were detected, which indicates some partial deglycosylation. Purification process does not alter the peptide mapping profile and purification process stresses are not the cause of peptide mapping noncompliance.

  12. Aplicación de una técnica de Cromatografía de Exclusión molecular para la purificación de ADN en plantas de Coffea sp. APPLICATION OF A TECHNIQUE OF MOLECULAR EXCLUSION CHROMATOGRAPHY FOR THE PURIFICATION OF DNA FROM Coffea sp. PLANTS

    Directory of Open Access Journals (Sweden)

    Ana María García Cepero


    Full Text Available Uno de los mayores inconvenientes en la extracción y purificación de biomoléculas a partir de plantas del género Coffea, es un alto contenido de polifenoles y compuestos tánicos. En el presente artículo se describe una metodología que permite obtener ADN de alta pureza. La extracción del ADN del homogeneizado de tejido foliar en siete genotipos de Coffea sp., se realizó mediante la técnica citada por Chaparro (1993 y su purificación se logró mediante cromatografía de exclusión molecular sobre una fase estacionaria de Sephacryl S-1000. Los resultados muestran que la alta eficiencia de separación de ARN degradado, proteínas, pigmentos y compuestos que absorben entre 220 y 300 nm, permiten obtener un ADN de alta pureza a juzgar por los datos espectrofotométricos y electroforéticos.One of the greatest difficulties in extracting and purifying biomolecules from plants in the genus Coffea is the high polyphenol and tannin contents. In this study a methodology is described that allows obtaining high purity DNA from leaf tissues of seven genotypes of Coffea sp. by means of the technique desribed by Chaparro (1993 and its further purification was achieved by molecular exclusion chromatography on Sephacryl S-1000 (Pharmacia. The results showed that the high separation efficiency of degraded RNA, proteins, pigments, and other compounds that absorb between 220 and 300 nm allowed obtaining high purity DNA as judged by the spectophometric and electroforetic data.

  13. Gas purification process. Verfahren zur Gasreinigung

    Energy Technology Data Exchange (ETDEWEB)

    Hoelter, H.; Gresch, H.; Igelbuescher, H.; Dewert, H.


    To avoid the problems of reheating in a wet process as well as the problems of higher gas supply in a dry process, the invention proposes to separate the raw gas in two component currents, one of which undergoes wet purification while the other is led through a dry purification process. The two component currents are mixed before entering the stack. The dry chemisorption masses added in substoichiometric doses are treated in a milk-of-lime processing stage, after which the reacted and non-reacted chemisorption masses are treated by wet purification and then by oxidation.

  14. Evaluation of automated and manual DNA purification methods for detecting Ricinus communis DNA during ricin investigations. (United States)

    Hutchins, Anne S; Astwood, Michael J; Saah, J Royden; Michel, Pierre A; Newton, Bruce R; Dauphin, Leslie A


    In April of 2013, letters addressed to the President of United States and other government officials were intercepted and found to be contaminated with ricin, heightening awareness about the need to evaluate laboratory methods for detecting ricin. This study evaluated commercial DNA purification methods for isolating Ricinus communis DNA as measured by real-time polymerase chain reaction (PCR). Four commercially available DNA purification methods (two automated, MagNA Pure compact and MagNA Pure LC, and two manual, MasterPure complete DNA and RNA purification kit and QIAamp DNA blood mini kit) were evaluated. We compared their ability to purify detectable levels of R. communis DNA from four different sample types, including crude preparations of ricin that could be used for biological crimes or acts of bioterrorism. Castor beans, spiked swabs, and spiked powders were included to simulate sample types typically tested during criminal and public health investigations. Real-time PCR analysis indicated that the QIAamp kit resulted in the greatest sensitivity for ricin preparations; the MasterPure kit performed best with spiked powders. The four methods detected equivalent levels by real-time PCR when castor beans and spiked swabs were used. All four methods yielded DNA free of PCR inhibitors as determined by the use of a PCR inhibition control assay. This study demonstrated that DNA purification methods differ in their ability to purify R. communis DNA; therefore, the purification method used for a given sample type can influence the sensitivity of real-time PCR assays for R. communis. Published by Elsevier Ireland Ltd.

  15. Cloning, expression and purification of orthologous membrane proteins: a general protocol for preparation of the histidine sensor kinase ETR1 from different species. (United States)

    Classen, Elisa; Groth, Georg


    Orthologous proteins do not necessarily share the same function in all species and those sharing the same function might employ a modified catalytic mechanism. Thus, comparative analysis of homologous or orthologous proteins from different organisms can provide detailed information on the function and the mechanism of an entire protein family. The sensor kinase ETR1 from Arabidopsis thaliana has been well characterized by genetic, physiological and biochemical studies. However, as further model plants are coming into focus for plant hormone research, a general protocol for isolation and purification of orthologous ETR1 proteins seems instrumental for a detailed molecular analysis of this protein family. In this study, we describe the native purification of recombinant ETR1 from Arabidopsis thaliana by mild solubilization with the zwitter-ionic detergent Fos-Choline-14 and single-step purification by immobilized metal ion affinity chromatography. The same protocol was successfully applied for the purification of the orthologous proteins from the moss Physcomitrella patens subsp. patens and the tomato Lycopersicon esculentum. The successful transfer of the purification protocol to proteins of the same family which share sequence identity of 63-80% only suggests that this protocol presents a general purification strategy which is likely to apply also to the purification of other members of the sensor histidine kinase family.

  16. Chromosome aberration assays in Allium

    Energy Technology Data Exchange (ETDEWEB)

    Grant, W.F.


    The common onion (Allium cepa) is an excellent plant for the assay of chromosome aberrations after chemical treatment. Other species of Allium (A. cepa var. proliferum, A. carinatum, A. fistulosum and A. sativum) have also been used but to a much lesser extent. Protocols have been given for using root tips from either bulbs or seeds of Allium cepa to study the cytological end-points, such as chromosome breaks and exchanges, which follow the testing of chemicals in somatic cells. It is considered that both mitotic and meiotic end-points should be used to a greater extent in assaying the cytogenetic effects of a chemical. From a literature survey, 148 chemicals are tabulated that have been assayed in 164 Allium tests for their clastogenic effect. Of the 164 assays which have been carried out, 75 are reported as giving a positive reaction, 49 positive and with a dose response, 1 positive and temperature-related, 9 borderline positive, and 30 negative; 76% of the chemicals gave a definite positive response. It is proposed that the Allium test be included among those tests routinely used for assessing chromosomal damage induced by chemicals.

  17. Identification and purification of novel chlorogenic acids in Artemisia annua L

    Directory of Open Access Journals (Sweden)

    Wenwen Zhao


    Full Text Available Present work has been carried out to study the identification and purification of chlorogenic acids in Artemisia annua L. Thirty-six chlorogenic acids were identified from this plant. Among these fifteen viz. two monocaffeoylquinic acids (Mr354, five dicaffeoylquinic acids (Mr516, one feruloylquinic acid (Mr368, three caffeoylferuloylquinic acids (Mr530, two ferulylquinic acids (Mr544, one dimethoxy-cinnamoylquinic acid (Mr382 and one p-coumaroylquinic acid (Mr338 were reported first time in present study by LC/MSn . Cis-isomers of these chlorogenic acids were also identified. Furthermore, column chromatography was used for the separation and purification of these chlorogenic acid; by the use of petroleum ether and ethyl acetate decolorization methods as mentioned in the literature, thus separation and purification process carried out at the same time. Polyamide and dextran were also used to purify Dicaffeoylquinic acid and purity level reached 85.7% with a yield of 53.4% after the secondary purification by Sephadex LH-20. Result of study revealed that A. annua can not only used for the production of artemisinin, but also yielding different kinds of chlorogenic acids, thus making comprehensive utilization of this plant.

  18. Purification

    DEFF Research Database (Denmark)

    Andersen, Astrid Oberborbeck


    —was treated before it was returned to the river where it continues its flow downstream towards cultivated fields and, finally, into the Pacific Ocean. It takes specialized knowledge and manifold technologies to manage water and sustain life in Arequipa, and engineers are central actors for making water flow......In Arequipa, Peru’s second largest city, engineers work hard to control water flows and provide different sectors with clean and sufficient water. In 2011, only 10 percent of the totality of water used daily by Arequipa’s then close to 1 million people—in households, tourism, industry, and mining...

  19. Purification of highly chlorinated dioxins degrading enzyme

    Energy Technology Data Exchange (ETDEWEB)

    Ishii, K.; Furuichi, T.; Koike, K.; Kuboshima, M. [Hokkaido Univ. (Japan). Division of Environment Resource Engineering, Graduate School of Engineering


    Soil contamination caused by dioxins in and around sites of incinerators for municipal solid waste (MSW) is a concern in Japan. For example, scattering wastewater from a wet gas scrubber at an MSW incinerator facility in Nose, Osaka caused soil and surface water contamination. The concentration of dioxins in the soil was about 8,000 pg-TEQ/g. Other contamination sites include soils on which fly ash has been placed directly or improperly stored and landfill sites that have received bottom and fly ash over a long period. Some countermeasures are required immediately at these dioxins-contaminated sites. We have previously developed bioreactor systems for dioxin-contaminated water and soil. We have shown that a fungus, Pseudallescheria boydii (P. boydii), isolated from activated sludge treating wastewater that contained dioxins, has the ability to degrade highly chlorinated dioxins. A reaction product of octachlorinated dibenzo-p-dioxin (OCDD) was identified as heptachlorinated dibenzo-p-dioxin. Therefore, one of the pathways for degradation of OCDD by this fungus was predicted to be as follows: OCDD is transformed by dechlorination and then one of the remaining aromatic rings is oxidized. To apply P. boydii to on-site technologies (e.g., bioreactor systems), as well as in situ technologies, enzyme treatment using a dioxin-degrading enzyme from P. boydii needs to be developed because P. boydii is a weak pathogenic fungus, known to cause opportunistic infection. As a result, we have studied enzyme purification of nonchlorinated dioxin, namely, dibenzo-pdioxin (DD). However, we did not try to identify enzymes capable of degrading highly chlorinated dioxins. This study has elucidated a method of enzyme assay for measuring OCDD-degrading activity, and has attempted to purify OCDD-degrading enzymes from P. boydii using enzyme assay. In addition, as first step toward purifying 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD), 2,3,7,8-TCDD degradation tests were carried out

  20. 含硫天然气净化厂硫化氢泄漏分析及对策%Study of hydrogen sulfide leakage and dispersion in sour gas purification plants and countermeasures

    Institute of Scientific and Technical Information of China (English)

    高少华; 邹兵; 严龙; 张贺; 王振


    Processing facilities with higher risk of H2S leakage were determined through analyzing purifying process and various causes that may result in leakage from one of the sour gas plants in Northeastern Sichuan. The feed gas in the desulphurization units and acid gas in the sulfur recovery units , mainly consisting of H2S and CO2 ,were selected for the simulation through comparing the process pressures, flow rates and material compositions. Weather conditions and terrains around the plant were studied. According to the statistics of local meteorological data during the last 5 years, low wind speed and cloudy weather were typical weather conditions, and Dl. 5m/s was selected for calculation of hydrogen sulfide dispersion at the plant. Surface roughness length recommended for small refineries by American Petroleum Institute was adopted. The Immediately Dangerous to Life or Health (IDLH ) zones resulted from the hazardous gas dispersions were calculated with PHAST based on three different leakage holes with lmin, 5min and 30min. The IDLH downwind distances range from 41m to 1190m with the farthest one resulting from the large hole release of feed gas from the desulphurization units under the typical weather. The effects of wind speeds and atmospheric stabilities on the H2S dispersion were also simulated and discussed with small hole release. Some suggestions were made for setting online leak detection and interlock systems in order to reduce the risk of H2S leakage It has instructive meanings in avoiding or reducing casualties due to hydrogen sulfide poisoning.%以川东北某含硫天然气净化厂为对象,通过分析该净化厂的处理工艺及可能造成泄漏的各种原因,确定了硫化氢泄漏危险较高的生产单元.通过工艺压力、流量、物料组分的比对,选取了脱硫单元原料气和硫磺回收单元酸性气作为模拟泄漏物料.对该厂所在地的气象条件和厂区的地形地貌进行了调查,净化厂当地近5年风速


    Directory of Open Access Journals (Sweden)

    R. Satheeskumar


    Full Text Available Partial purification and characterization of alkalophilic protease production from Pseudomonas aeruginosa was isolated from the gut of marine and coastal waters shrimp Penaeus monodon. The protease production was assayed in submerged fermentation to produce maximum protease activity (423 ± 0.09 U/ml. The enzyme was precipitated with ammonium sulphate and partially purified by ion exchange chromatography through DEAE Sephadex A-50 column. In 10th fraction showed maximum protease activity (734 ± 0.18 U/ml with increase in purification fold. The molecular weight of protease from Pseudomonas aeruginosa was recorded as 60 kDa. The stability of protease was tested at various pH and temperature; it showed maximum protease activity at pH-9 and temperature 50ºC. Among the various surfactants tested for enzyme stability, maximum activity was retained in poly ethylene glycol. The compatibility of protease enzyme with various commercial detergents; the enzyme retained maximum protease activity in tide. The results are indicated that all these properties make the bacterial proteases are most suitable for wide industrial applications.

  2. Reverse osmosis water purification system (United States)

    Ahlstrom, H. G.; Hames, P. S.; Menninger, F. J.


    A reverse osmosis water purification system, which uses a programmable controller (PC) as the control system, was designed and built to maintain the cleanliness and level of water for various systems of a 64-m antenna. The installation operates with other equipment of the antenna at the Goldstone Deep Space Communication Complex. The reverse osmosis system was designed to be fully automatic; with the PC, many complex sequential and timed logic networks were easily implemented and are modified. The PC monitors water levels, pressures, flows, control panel requests, and set points on analog meters; with this information various processes are initiated, monitored, modified, halted, or eliminated as required by the equipment being supplied pure water.

  3. Rehabilitation Scheme and Water Purification Effect of Advanced Treatment Processes for Songjiang No.2 Water Treatment Plant%松江二水厂深度处理改造方案及净水效果

    Institute of Scientific and Technical Information of China (English)

    冯钧; 陶明; 徐建平


    In order to meet the new drinking water standards, the Songjiang No.2 Water Treatment Plant reconstructed the conventional horizontal-flow sedimentation basin into a short horizontal-flow sedimentation basin, an inclined-tube sedimentation basin and an up-flow GAC filter. To realize the goal of land and energy saving, an ozone-BAC advanced treatment system was designed and established. The effluent quality was improved.%为使出厂水达到新颁(GB 5749-2006)的要求,松江自来水公司二水厂将水平沉淀池改造为短水平沉淀池、斜管沉淀池及上向流活性炭滤池,利用两组沉淀池之间的空间安装臭氧设备.在节能、节地条件下,实现了臭氧生物活性炭水处理工艺,提高了出厂水水质.

  4. Research on Wastewater Purification and Its Recycling in a Lead-Zinc Ore Beneficiation Plant%铅锌选矿厂废水净化回用研究

    Institute of Scientific and Technical Information of China (English)



    介绍了采用“中和-机械混合反应-斜管沉淀-活性炭吸附”对铅锌选矿厂选矿废水进行处理后回用的方法,运行结果表明,采用该工艺处理后废水达到选矿工艺用水标准要求,由此不仅节约了大量新水,而且处理效率高,运行成本低,创造了良好的经济效益、社会效益和环境效益.%The method of wastewater reuse after treating wastewater from a lead-zinc ore benefieiation plant covering neutralisation, mix reaction by machinery, inclined lube sedimentation and activated carbon adsorption was introduced. The running results show that wastewater treated by the proposal technology has met the requirement of water standards for mineral processing technology, which saves a large amount of fresh water, and has created favorable economic, social and environmental benefits.

  5. Liquid membrane purification of biogas

    Energy Technology Data Exchange (ETDEWEB)

    Majumdar, S.; Guha, A.K.; Lee, Y.T.; Papadopoulos, T.; Khare, S. (Stevens Inst. of Tech., Hoboken, NJ (United States). Dept. of Chemistry and Chemical Engineering)


    Conventional gas purification technologies are highly energy intensive. They are not suitable for economic removal of CO{sub 2} from methane obtained in biogas due to the small scale of gas production. Membrane separation techniques on the other hand are ideally suited for low gas production rate applications due to their modular nature. Although liquid membranes possess a high species permeability and selectivity, they have not been used for industrial applications due to the problems of membrane stability, membrane flooding and poor operational flexibility, etc. A new hollow-fiber-contained liquid membrane (HFCLM) technique has been developed recently. This technique overcomes the shortcomings of the traditional immobilized liquid membrane technology. A new technique uses two sets of hydrophobic, microporous hollow fine fibers, packed tightly in a permeator shell. The inter-fiber space is filled with an aqueous liquid acting as the membrane. The feed gas mixture is separated by selective permeation of a species through the liquid from one fiber set to the other. The second fiber set carries a sweep stream, gas or liquid, or simply the permeated gas stream. The objectives (which were met) of the present investigation were as follows. To study the selective removal of CO{sub 2} from a model biogas mixture containing 40% CO{sub 2} (the rest being N{sub 2} or CH{sub 4}) using a HFCLM permeator under various operating modes that include sweep gas, sweep liquid, vacuum and conventional permeation; to develop a mathematical model for each mode of operation; to build a large-scale purification loop and large-scale permeators for model biogas separation and to show stable performance over a period of one month.

  6. 锰污染土壤渗漏液与径流生态拦截净化系统的植物筛选%Screening of plant species for establishing an retention and purification ecosystem of soil infiltration water and surface runoff in manganese polluted area

    Institute of Scientific and Technical Information of China (English)

    陈星; 文仕知; 陈永华; 郝君; 刘凯; 吴子剑


    Screening of plant species were carried out for establishing an ecosystem for retention and purification of soil infiltration water and surface runoff in Xiangtan manganese polluted area. The results obtained from a five month plant growth period indicate that mushroom grass had a very low survival rate while Arundo donax var. versicolor and Acorus calamus Linn had a negative value in its biomass increment. In comparison, the other nine plant species, Thalia dealbata, Boehmeria, Canna warscewiezii A. Dietr, Phragmites australis, Typha orientalis Presl, Pontederia cordala, Nerium oleander, Pontederia cordata, Sofistem bulrush and Iris germanica grew well in the manganese polluted sites. The manganese contents in shoots of the nine plant species were all more than 1000 mg/kg and their zinc, copper and cadmium contents were also relatively high, with the ratio of the metal content in above-ground tissues to that in roots being greater than 1. In contrast, the above-ground tissue to root ratio of zinc, copper, manganese and cadmium contents in A. calamus and that of zinc, copper, and cadmium contents in A. donax var versicolor were lower than 1, suggesting that the metal accumulation in roots due to weak heavy metal transfer abilities of these species had led to poisoning effects on the pant growth. The highest manganese uptake in above-ground tissues of Boehmeria reached 217.8 mg per plant. The next uptake value was given by T. dealbata, Boehmeria, followed in turn by C. warscewiezii, Dietr, P. australis, P. cordata and S. bulrush.%为建立锰污染土壤渗漏液和径流收集处理系统,在湘潭锰矿废弃地开展了植物筛选试验.5个月植物生长的试验结果表明,香菇草成活率低,花叶芦竹、菖蒲生长量下降,而再力花、苎麻、紫叶美人蕉、芦苇、香蒲、夹竹桃、梭鱼草、水葱和德国鸢尾长势良好,其地上部分锰的含量多高于1 000 mg/kg,锌、铜、镉的含量也相对较高,锰含量地上

  7. Purification of rhamnolipid using colloidal magnetic nanoparticles

    African Journals Online (AJOL)



    Jul 6, 2009 ... separation and purification of bio-molecules, particularly, .... of model Rhamnolipid by ion exchange processes. The ..... dynamic diameter of the particles is 35.6 nm. ... solubility of rhaminolipid was decreased and amount of.

  8. Accelerated purification of colloidal silica sols (United States)

    Bahnsen, E. B.; Garofalini, S.; Pechman, A.


    Accelerated purification process for colloidal sols using heat/deionization scheme, sharply reduces waiting time between deionization cycles from several months to a few days. Process produces same high purity silica sols as conventional methods.

  9. Solid State Air Purification System Project (United States)

    National Aeronautics and Space Administration — The purpose of this proposed research is to develop a new air purification system based on a liquid membrane, capable of purifying carbon dioxide from air in a far...

  10. Affinity purification of recombinant human plasminogen activator ...

    African Journals Online (AJOL)

    Purpose: To develop processes for effective isolation and purification of recombinant human plasminogen ... three hybridoma strains were superior for producing PR-mAbs (C1, C4, C8). ..... characterization of a polyol- responsive monoclonal.

  11. Extracorporeal blood purification in burns: a review. (United States)

    Linden, Katharina; Stewart, Ian J; Kreyer, Stefan F X; Scaravilli, Vittorio; Cannon, Jeremy W; Cancio, Leopoldo C; Batchinsky, Andriy I; Chung, Kevin K


    A prolonged and fulminant inflammatory state, with high levels of pro- and anti-inflammatory mediators, is seen after extensive thermal injury. Blood purification techniques including plasma exchange, continuous venovenous hemofiltration, and adsorbing membranes have the potential to modulate this response, thereby improving outcomes. This article describes the scientific rationale behind blood purification in burns and offers a review of literature regarding its potential application in this patient cohort.

  12. Purification system in Puglia region. Control of efficiency of the treatment plants. State-of-the-art and perspectives; Il sistema depurativo in Puglia. Controllo dell'efficienza degli impianti di trattamento, stato dell'arte e prospettive

    Energy Technology Data Exchange (ETDEWEB)

    D' Elia, E. [Naples Univ. Federico 2., Naples (Italy). Dipt. di Protezione Idraulica del Territorio; Di Marino, R. [Aquedotto Pugliese Spa, Bari (Italy)


    This report was written to provide information regarding the depuration system and the final destinations of urban wastewater in the Puglia Region and arises form the need to provide a fully detailed analysis of the situation so that proper action may be taken by the Prefect of Bari, Delegate Commissioner on behalf of the Italian Government for the current state of social, economic and environmental emergency in the Region. The report analyses the current status of the entire system, the territorial network of distribution as related to the performance of pollution reduction, the sludge digestion systems utilized and the types of final discharge. Finally, the procedures currently in use of verifying the efficiency of depuration are described and the principal results which indirectly provide an accounting of the reliability of the project-criteria as well as the present state of functionality of the treatment plants. [Italian] Questa nota si prefigge lo scopo di fornire un contributo alla conoscenza del sistema depurativo e dei recapiti finali delle acque reflue urbane che attualmente si registra nella Regione Publia e nasce dall'esigenza di disporre di un esauriente quadro conoscitivo, utile all'azione del Prefetto di Bari, Commissario di Governo per lo stato di Emergenza socio-economico-ambientale in cui versa la Regione. Viene descritta la situazione attuale dell'intero sistema e la sua distribuzione sul territorio, in connessione con il rendimento di abbattimento del carico inquinante, del tipo di digestione ei fanghi e della natura dei recapiti finali. Vengono infine descritte le procedure correnti di verifica dell'efficienza di depurazione e ne vengono forniti i principali risultati che danno indirettamente conto della attendibilita' dei criteri di progettazione e dell'attuale stato di funzionamento degli impianti di trattamento.

  13. Efficient expression and purification of biologically active human cystatin proteins. (United States)

    Chauhan, Sakshi; Tomar, Raghuvir S


    Cystatins are reversible cysteine protease inhibitor proteins. They are known to play important roles in controlling cathepsins, neurodegenerative disease, and in immune system regulation. Production of recombinant cystatin proteins is important for biochemical and function characterization. In this study, we cloned and expressed human stefin A, stefin B and cystatin C in Escherichia coli. Human stefin A, stefin B and cystatin C were purified from soluble fraction. For cystatin C, we used various chaperone plasmids to make cystatin C soluble, as it is reported to localize in inclusion bodies. Trigger factor, GroES-GroEL, DnaK-DnaJ-GrpE chaperones lead to the presence of cystatin C in the soluble fraction. Immobilized metal affinity chromatography, glutathione sepharose and anion exchange chromatography techniques were employed for efficient purification of these proteins. Their biological activities were tested by inhibition assays against cathepsin L and H3 protease.

  14. Microbead agglutination based assays

    KAUST Repository

    Kodzius, Rimantas


    We report a simple and rapid room temperature assay for point-of-care (POC) testing that is based on specific agglutination. Agglutination tests are based on aggregation of microbeads in the presence of a specific analyte thus enabling the macroscopic observation. Such tests are most often used to explore antibody-antigen reactions. Agglutination has been used for protein assays using a biotin/streptavidin system as well as a hybridization based assay. The agglutination systems are prone to selftermination of the linking analyte, prone to active site saturation and loss of agglomeration at high analyte concentrations. We investigated the molecular target/ligand interaction, explaining the common agglutination problems related to analyte self-termination, linkage of the analyte to the same bead instead of different microbeads. We classified the agglutination process into three kinds of assays: a two- component assay, a three-component assay and a stepped three- component assay. Although we compared these three kinds of assays for recognizing DNA and protein molecules, the assay can be used for virtually any molecule, including ions and metabolites. In total, the optimized assay permits detecting analytes with high sensitivity in a short time, 5 min, at room temperature. Such a system is appropriate for POC testing.

  15. Colorimetric protein assay techniques. (United States)

    Sapan, C V; Lundblad, R L; Price, N C


    There has been an increase in the number of colorimetric assay techniques for the determination of protein concentration over the past 20 years. This has resulted in a perceived increase in sensitivity and accuracy with the advent of new techniques. The present review considers these advances with emphasis on the potential use of such technologies in the assay of biopharmaceuticals. The techniques reviewed include Coomassie Blue G-250 dye binding (the Bradford assay), the Lowry assay, the bicinchoninic acid assay and the biuret assay. It is shown that each assay has advantages and disadvantages relative to sensitivity, ease of performance, acceptance in the literature, accuracy and reproducibility/coefficient of variation/laboratory-to-laboratory variation. A comparison of the use of several assays with the same sample population is presented. It is suggested that the most critical issue in the use of a chromogenic protein assay for the characterization of a biopharmaceutical is the selection of a standard for the calibration of the assay; it is crucial that the standard be representative of the sample. If it is not possible to match the standard with the sample from the perspective of protein composition, then it is preferable to use an assay that is not sensitive to the composition of the protein such as a micro-Kjeldahl technique, quantitative amino acid analysis or the biuret assay. In a complex mixture it might be inappropriate to focus on a general method of protein determination and much more informative to use specific methods relating to the protein(s) of particular interest, using either specific assays or antibody-based methods. The key point is that whatever method is adopted as the 'gold standard' for a given protein, this method needs to be used routinely for calibration.

  16. Expression, Purification and Activity Assay of the Full-length and Truncated Human Cystathionine β-Synthase%人胱硫醚β-合酶及其截短型片段的表达、纯化和活性测定

    Institute of Scientific and Technical Information of China (English)

    牛卫宁; 羊梦林; 曹珊珊; 许乐; 钦传光


    used. From a 1L culture, some 15.2 mg of purified CBS protein at 95% purity as judged by SDS-PAGE could be abtained, and the purified enzyme had a specific activity of 143 unit/mg protein. S-adenosylmethionine(AdoMet) activates the CBS enzyme by as much as 5. 1-fold in the presence of 1 mmol/L AdoMet with a specific activity of 735 unit/mg protein. Additionally, the recombinant E. Coli Rosetta (pETDuet-l-CBS1-413) strain which highly express truncated CBS(CBS1-413) gene was constructed. Using a His Trap Fast Flow affinity chromatography, the purity of recombinant CBS1-413 lacking the C-terminal regulatory domain reached 95% by one-step purification with the specific activity of 965 unit/mg. The productivity of the soluble CBS reached 12. 8mg/L and in 74. 3% overall yield. In addition, The expression and purification of recombinant cystathionine p-lyase (CBL) in E. Coli were described. The present study established a novel method, which relies on CBL as coupling enzyme, for detemination of CBS activity based on the color reaction between pyruvate and 2,4-dinitrophenylhydrazine.

  17. Production of nodulation factors by Rhizobium meliloti: fermentation, purification and characterization of glycolipids. (United States)

    Kohring, B; Baier, R; Niehaus, K; Pühler, A; Flaschel, E


    Lipooligosaccharides, synthesized by soil bacteria of the genera Rhizobium, are known to have multifunctional effects on a wide variety of plants as signal substances in symbiosis initiation, cell response elicitation and growth regulation. These so called nodulation (Nod-) factors represent interesting biotechnological products with respect to fundamental studies of symbiotic interactions as well as for potential applications. Therefore, a batch fermentation process on a scale of 30 l has been developed by means of the Rhizobium meliloti strain R.m. 1021 (pEK327) strongly overexpressing the genes for the synthesis of Nod factors. Induction by the flavone luteolin led to growth associated production of the lipooligosaccharides. Ultrafiltration was used for separating the biomass from the filtrate containing the extracellular Nod factors. Simultaneously, ultrafiltration reduced the amount of lipophilic substances, which would otherwise interfere with processes downstream. The second separation step consisted in adsorption on XAD-2, a nonspecific hydrophobic adsorptive resin. Adsorption of Nod factors was carried out by batch operation of a stirred tank. Desorption was performed by elution with methanol in a fixed bed column. A semi-preparative reversed phase HPLC (Polygoprep 100-30 C18) was chosen as the final purification step. The Nod factors were obtained after evaporation and lyophilization. Thus, about 600 mg of Nod factors were produced from 20 l of fermentation broth. The Nod factors produced by Rhizobium meliloti R.m. 1021 (pEK327) were identified by liquid secondary ion mass spectrometry and by reversed-phase HPLC as fluorescent derivatives of 2-aminobenzamide. The biological activity of the products was demonstrated by means of the root hair deformation (HAD-) assay.

  18. Unified Model of Purification Units in Hydrogen Networks

    Institute of Scientific and Technical Information of China (English)

    吴思东; 王彧斐; 冯霄


    Purification processes are widely used in hydrogen networks of refineries to increase hydrogen reuse. In refineries, hydrogen purification techniques include hydrocarbon, hydrogen sulfide and CO removal units. In addi-tion, light hydrocarbon recovery from the hydrogen source streams can also result in hydrogen purification. In order to simplify the superstructure and mathematical model of hydrogen network integration, the models of different pu-rification processes are unified in this paper, including mass balance and the expressions for hydrogen recovery and impurity removal ratios, which are given for all the purification units in refineries. Based on the proposed unified model, a superstructure of hydrogen networks with purification processes is constructed.

  19. Production, partial purification and characterization of feruloyl esterase by Aspergillus awamori in submerged fermentation. (United States)

    Fazary, Ahmed Eid; Ju, Yi-Hsu


    Microbial feruloyl esterases acting on plant cell wall polymers represent key tools for the degradation of plant cell wall. In this paper, we describe in detail the microbial production, partial purification and characterization of feruloyl esterase from a culture medium of Aspergillus awamori strain IFO4033 obtained from a crude hemicellulose preparation of wheat straw, corncobs and wheat germ. Feruloyl esterase was extracted using centrifugation and dialysis, and then purified by ion exchange chromatography and microfiltration to homogeneity, which was checked by SDSPAGE and isoelectric focusing-PAGE. Protein content and activity of the enzyme were measured in each step of extraction and purification. Biomass was determined by the dry weight method. pH and temperature optima of feruloyl esterase enzyme were also determined. The effects of culturing time, and carbon and nitrogen sources on enzyme production were systematically investigated. Finally, enzyme activities under different storage conditions were examined.

  20. Absolute nuclear material assay (United States)

    Prasad, Manoj K [Pleasanton, CA; Snyderman, Neal J [Berkeley, CA; Rowland, Mark S [Alamo, CA


    A method of absolute nuclear material assay of an unknown source comprising counting neutrons from the unknown source and providing an absolute nuclear material assay utilizing a model to optimally compare to the measured count distributions. In one embodiment, the step of providing an absolute nuclear material assay comprises utilizing a random sampling of analytically computed fission chain distributions to generate a continuous time-evolving sequence of event-counts by spreading the fission chain distribution in time.

  1. New research on bioregenerative air/water purification systems (United States)

    Johnson, Anne H.; Ellender, R. D.; Watkins, Paul J.


    For the past several years, air and water purification systems have been developed and used. This technology is based on the combined activities of plants and microorganisms as they function in a natural environment. More recently, researchers have begun to address the problems associated with indoor air pollution. Various common houseplants are currently being evaluated for their abilities to reduce concentrations of volatile organic compounds (VOCS) such as formaldehyde and benzene. With development of the Space Exploration Initiative, missions will increase in duration, and problems with resupply necessitates implementation of regenerative technology. Aspects of bioregenerative technology have been included in a habitat known as the BioHome. The ultimate goal is to use this technology in conjunction with physicochemical systems for air and water purification within closed systems. This study continued the risk assessment of bioregenerative technology with emphasis on biological hazards. In an effort to evaluate the risk for human infection, analyses were directed at enumeration of fecal streptococci and enteric viruses with the BioHome waste water treatment system.

  2. Simple optimization method for partitioning purification of hydrogen networks

    Directory of Open Access Journals (Sweden)

    W.M. Shehata


    Full Text Available The Egyptian petroleum fuel market is increasing rapidly nowadays. These fuels must be in the standard specifications of the Egyptian General Petroleum Corporation (EGPC, which required lower sulfur gasoline and diesel fuels. So the fuels must be deep hydrotreated which resulted in increasing hydrogen (H2 consumption for deeper hydrotreating. Along with increased H2 consumption for deeper hydrotreating, additional H2 is needed for processing heavier and higher sulfur crude slates especially in hydrocracking process, in addition to hydrotreating unit, isomerization units and lubricant plants. Purification technology is used to increase the amount of recycled hydrogen. If the amount of recycled hydrogen is increased, the amount of hydrogen that is sent to the furnaces with the off gas will decrease. In this work, El Halwagi et al. (2003 and El Halwagi (2012 optimization methods which are used for recycle/reuse integration systems have been extended to be used in the partitioning purification of hydrogen networks to minimize the hydrogen consumption and the hydrogen discharge. An actual case study and two case studies from the literature are solved to illustrate the proposed method.

  3. Phytoremediation of Anaerobic Digester Effluent for Water Purification and Production of Animal Feed


    Abdel E. Ghaly; H. A. Farag


    The application of phytoremediation for purification of an anaerobically treated dairy manure and production of forage crops was investigated. Four crops (two cereals and two grasses) were examined for their ability to grow hydroponically and to remove pollutants (nutrients) from dairy wastewater. The preliminary experiments showed that timothygrass and orchardgrass did not perform well as aquatic plants. Only 24 and 29% of the seeds germinated after 19-21 days giving a crop yield of 21 and 1...

  4. Partial purification and characterization of a Ca(2+)-dependent protein kinase from the green alga, Dunaliella salina (United States)

    Roux, S. J.


    A calcium-dependent protein kinase was partially purified and characterized from the green alga Dunaliella salina. The enzyme was activated at free Ca2+ concentrations above 10(-7) molar. and half-maximal activation was at about 3 x 10(-7) molar. The optimum pH for its Ca(2+)-dependent activity was 7.5. The addition of various phospholipids and diolein had no effects on enzyme activity and did not alter the sensitivity of the enzyme toward Ca2+. The enzyme was inhibited by calmodulin antagonists, N-(6-aminohexyl)-1-naphthalene sulfonamide and N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide in a dose-dependent manner while the protein kinase C inhibitor, sphingosine, had little effect on enzyme activity up to 800 micromolar. Immunoassay showed some calmodulin was present in the kinase preparations. However, it is unlikely the kinase was calmodulin regulated, since it still showed stimulation by Ca2+ in gel assays after being electrophoretically separated from calmodulin by two different methods. This gel method of detection of the enzyme indicated that a protein band with an apparent molecular weight of 40,000 showed protein kinase activity at each one of the several steps in the purification procedure. Gel assay analysis also showed that after native gel isoelectric focusing the partially purified kinase preparations had two bands with calcium-dependent activity, at isoelectric points 6.7 and 7.1. By molecular weight, by isoelectric point, and by a comparative immunoassay, the Dunaliella kinase appears to differ from at least some of the calcium-dependent, but calmodulin and phospholipid independent kinases described from higher plants.

  5. Cyclodextrin purification with hollow fibers

    Energy Technology Data Exchange (ETDEWEB)

    Berthod, A. (Univ. de Lyon 1, Villeubranne Cedex (France)); Jin, Heng Liang,; Armstrong, D.W. (Univ. of Missouri, Rolla (USA))


    Cyclodextrins are cyclic 1-4 linked oligomers of {alpha}-D-glucopyranose prepared from starch hydrolysis through enzymatic reactions. Mixtures of the three main cyclodextrins (CD), {alpha}-, {beta}-, and {gamma}-CDs, are always produced. A possible facile purification process is proposed. Permeation through hollow fibers made of a perfluorinated ionomer membrane. Nafion type, is shown to be an effective way to separate {alpha}-CD from {beta}- and {gamma}-CD. {Alpha}-CD with 95% purity was obtained after permeation through a Nafion hollow fiber of an equimolar 0.02 M solution of the three CDs. The fiber had a 56 cm{sup 2}/cm{sup 3} surface area per volume ratio. Kinetic studies and continuous extraction experiments with a 2-m coiled fiber showed that it is possible to obtain a 11.5 g {alpha}-CD solution with 92.4% purity or a 0.6 g {alpha}-CD solution with 97.2% purity, depending on the flow rate. The transport of CDs through the membrane could be due to moving water pools inside the ionomer. The small {alpha}-CD fits easily in such pools when the large {beta}- and {gamma}-CDs are excluded by steric hindrance. Temperature raises increased the permeation rates while decreasing the selectivity. The process could be scaled-up associating hollow fibers in bundle.

  6. Cloning, high-level expression, purification and characterization of a ...

    African Journals Online (AJOL)

    Cloning, high-level expression, purification and characterization of a staphylokinase variant, SakøC, ... African Journal of Biotechnology ... Hence in this study, we reported the cloning, high-level expression, purification and characterization of ...

  7. Isolation, purification and properties of lipase from Pseudomonas ...

    African Journals Online (AJOL)

    Isolation, purification and properties of lipase from Pseudomonas aeruginosa. ... medium agar containing Tween 80 or olive oil as the only source of carbon. ... through the purification procedure of ammonium sulphate precipitation and diethyl ...

  8. 高含硫气田开发酸污对天然气净化厂输电线路安全影响研究及防护措施优选%Optimization of protective measures and safety impact study of high sulfur gas field acid pollution on the electric transmission line of natural gas purification plant

    Institute of Scientific and Technical Information of China (English)

    王和琴; 陈惟国; 苗辉; 周培利


    基于高含硫气田开发过程中的天然气净化厂尾气排放特性及在大气中浓度分布的预测,以普光气田为例,就酸性气体对220 kV 线路绝缘性能分析,分别测试了模拟酸雾环境中污秽绝缘子以及空气的绝缘性能。结果表明,在盐雾的作用下,绝缘子的闪络电压下降了约20%,雾的pH值为3时,空气的绝缘强度下降了约15.1%,pH值等于3的酸雾与II级空气污染相结合,空气的绝缘强度大约下降了23.2%。在pH值等于3的酸雨酸雾环境下,线路的耐雷水平下降了约20%。结合实验室分析结果和普光气田的实际情况,分析了酸性气田输电线路的防护措施。%Based on the gas emission characteristics of natural gas purification plant and con-centration distribution prediction in atmosphere during the development process of high sour gas filed ,insulation performance analysis of acid gases on the 220 kV line was conducted ,taking Pu-guang gas field as example .Insulating property of polluted insulator and the air under simulated acid mist environment were measured respectively .The results showed that flashover voltage of insulator dropped about 20% under the action of salt mist .Insulating strength of the air dropped about 15 .1% when the pH value of mist was 3 .After the combination of acid mist (pH=3) and class II air pollution ,insulating strength of the air dropped about 23 .2% .Lightning withstand level of electric transmission line decreased 20% approximately under acid rain and acid mist envi-ronment with pH value of 3 .Combined the laboratory analysis results and practical situation in Puguang gas field ,the protective measures of electric transmission line in acid gas field were ana-lyzed .

  9. Photocatalytic materials and technologies for air purification. (United States)

    Ren, Hangjuan; Koshy, Pramod; Chen, Wen-Fan; Qi, Shaohua; Sorrell, Charles Christopher


    Since there is increasing concern for the impact of air quality on human health, the present work surveys the materials and technologies for air purification using photocatalytic materials. The coverage includes (1) current photocatalytic materials for the decomposition of chemical contaminants and disinfection of pathogens present in air and (2) photocatalytic air purification systems that are used currently and under development. The present work focuses on five main themes. First, the mechanisms of photodegradation and photodisinfection are explained. Second, system designs for photocatalytic air purification are surveyed. Third, the photocatalytic materials used for air purification and their characteristics are considered, including both conventional and more recently developed photocatalysts. Fourth, the methods used to fabricate these materials are discussed. Fifth, the most significant coverage is devoted to materials design strategies aimed at improving the performance of photocatalysts for air purification. The review concludes with a brief consideration of promising future directions for materials research in photocatalysis. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Novel peptide ligand with high binding capacity for antibody purification

    DEFF Research Database (Denmark)

    Lund, L. N.; Gustavsson, P. E.; Michael, R.


    Small synthetic ligands for protein purification have become increasingly interesting with the growing need for cheap chromatographic materials for protein purification and especially for the purification of monoclonal antibodies (mAbs). Today, Protein A-based chromatographic resins are the most ......-aggregated IgG, indicating that the ligand could be used both as a primary purification step of IgG as well as a subsequent polishing step. (C) 2012 Elsevier B.V. All rights reserved....

  11. Fibulin-1 purification from human plasma using affinity chromatography on Factor H-Sepharose. (United States)

    DiScipio, Richard G; Liddington, Robert C; Schraufstatter, Ingrid U


    A method is reported to purify Fibulin-1 from human plasma resulting in a 36% recovery. The steps involve removal of the cryoglobulin and the vitamin K dependent proteins followed by polyethylene glycol and ammonium sulfate precipitations, DEAE-Sephadex column chromatography and finally Factor H-Sepharose affinity purification. The procedure is designed to be integrated into an overall scheme for the isolation of over 30 plasma proteins from a single batch of human plasma. Results from mass spectroscopy, SDS-PAGE, and Western blotting indicate that human plasma Fibulin-1 is a single chain of the largest isotype. Functional binding assays demonstrated calcium ion dependent interaction of Fibulin-1 for fibrinogen, fibronectin, and Factor H. The procedure described is the first to our knowledge that enables a large scale purification of Fibulin-1 from human plasma.

  12. Argatroban-coupled Affi-Gel matrix for the purification of thrombin from plasma. (United States)

    Lefkowitz, Jerry B


    Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay. None of the available purification methods easily deals with this subject. The procedure described in the present paper uses a readily available pharmaceutical agent, argatroban, to construct an affinity matrix. Argatroban has a high affinity for thrombin and its thrombin binding is reversible. Prothrombin derived from a Ba(2+) precipitate of human plasma is used as the starting material. The crude prothrombin can be bulk activated to thrombin using taipan-snake (Oxyuranus scutellatus) venom and bound to the argatroban-coupled matrix without further processing steps. The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis. This purification scheme is rapid, yielding purified thrombin within 2 days.

  13. Purification of histidine-tagged T4 RNA ligase from E. coli. (United States)

    Wang, Qing S; Unrau, Peter J


    Here we report the construction of a histidine-tagged T4 RNA ligase expression plasmid (pRHT4). The construct, when overexpressed in BL21 (DE3) cells, allows the preparation of large quantities of T4 RNA ligase in high purity using only a single purification column. The histidine affinity tag does not inhibit enzyme function, and we were able to purify 1-3 mg pure protein/g cell pellet. A simple purification procedure ensures that the enzyme is de-adenylated to levels comparable to those found for many commercial preparations. The purified protein has very low levels of RNase contamination and functioned normally in a variety of activity assays.

  14. Purification of capping protein using the capping protein binding site of CARMIL as an affinity matrix. (United States)

    Remmert, Kirsten; Uruno, Takehito; Hammer, John A


    Capping protein (CP) is a ubiquitously expressed, heterodimeric actin binding protein that is essential for normal actin dynamics in cells. The existing methods for purifying native CP from tissues and recombinant CP from bacteria are time-consuming processes that involve numerous conventional chromatographic steps and functional assays to achieve a homogeneous preparation of the protein. Here, we report the rapid purification of Acanthamoeba CP from amoeba extracts and recombinant mouse CP from E. coli extracts using as an affinity matrix GST-fusion proteins containing the CP binding site from Acanthamoeba CARMIL and mouse CARMIL-1, respectively. This improved method for CP purification should facilitate the in vitro analysis of CP structure, function, and regulation.

  15. 21 CFR 876.5665 - Water purification system for hemodialysis. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Water purification system for hemodialysis. 876... SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5665 Water purification system for hemodialysis. (a) Identification. A water purification system for hemodialysis is...

  16. 煤基氮肥甲醇生产中深度净化技术综述(上)%Summary of deep purification technology in coal-based nitrogen fertilizer methanol production(Part Ⅰ)

    Institute of Scientific and Technical Information of China (English)



    简要介绍煤基氮肥、甲醇生产中气体深度净化技术,其中涉及中小型合成氨厂的深度净化技术,合成氨和甲醇厂的脱硫脱碳一体化净化技术,大型甲醇厂的深度净化技术,液氮洗净化技术,布朗深冷净化技术,焦炉煤气的深度净化技术等。%The paper briefly introduces deep purification technology in coal-based nitrogen fertilizer, methanol production, which involves deep purification technology in small and medium-sized ammonia plants, desulfurization integrated purification technology in ammonia and methanol plants, deep purification technology in large-scale methanol plants, purification technology in liquid ammonia washing, Brown cryogenic purification technology, deep purification technology in coke oven gas and so on.

  17. Purification and Characterization of Tryptophan Hydroxylase

    DEFF Research Database (Denmark)

    Haahr, Lærke Tvedebrink

    This thesis deals with the purification and characterization of the iron-containing enzyme tryptophan hydroxylase (TPH). TPH exists in two isoforms, called TPH1 and TPH2. Each isoform consists of threestructural distinct domains: the regulatory, the catalytic and the tetramerization domain. TPH...... of this project was to developpurification methods for full-length TPH1 and TPH2 as well as to characterize purified TPH variants. A successful purification method for full-length human TPH1 (hTPH1) was developed, which resulted in pure, active and stable protein. The method includes affinity-purification using....... The crystallization procedure for the catalytic domain of gallus gallus TPH1 (cgTPH1) was optimized to faster crystal growth by addition of tryptophan and incubation at room temperature. Crystals without imidazole in the crystallization conditions could be obtained. The solved structures were however of poor quality...

  18. Reconsidering Rapid Qubit Purification by Feedback

    CERN Document Server

    Wiseman, H M


    This paper reconsiders the properties of a scheme for the rapid purification of the quantum state of a qubit, proposed recently in Jacobs 2003 Phys. Rev. A67 030301(R). The qubit starts in a completely mixed state, and information is obtained by a continuous measurement. Jacobs' rapid purification protocol uses Hamiltonian feedback control to maximise the average purity of the qubit for a given time, with a factor of two increase in the purification rate over the no-feedback protocol. However, by re-examining the latter approach, we show that it mininises the average time taken for a qubit to reach a given purity. In fact, the average time taken for the no-feedback protocol beats that for Jacobs' protocol by a factor of two. We discuss how this is compatible with Jacobs' result, and the usefulness of the different approaches.

  19. Overview of the purification of recombinant proteins. (United States)

    Wingfield, Paul T


    When the first version of this unit was written in 1995, protein purification of recombinant proteins was based on a variety of standard chromatographic methods and approaches, many of which were described and mentioned throughout Current Protocols in Protein Science. In the interim, there has been a shift toward an almost universal usage of the affinity or fusion tag. This may not be the case for biotechnology manufacture where affinity tags can complicate producing proteins under regulatory conditions. Regardless of the protein expression system, questions are asked as to which and how many affinity tags to use, where to attach them in the protein, and whether to engineer a self-cleavage system or simply leave them on. We will briefly address some of these issues. Also, although this overview focuses on E.coli, protein expression and purification, other commonly used expression systems are mentioned and, apart from cell-breakage methods, protein purification methods and strategies are essentially the same.

  20. Purification of Carbon Nanotubes: Alternative Methods (United States)

    Files, Bradley; Scott, Carl; Gorelik, Olga; Nikolaev, Pasha; Hulse, Lou; Arepalli, Sivaram


    Traditional carbon nanotube purification process involves nitric acid refluxing and cross flow filtration using surfactant TritonX. This is believed to result in damage to nanotubes and surfactant residue on nanotube surface. Alternative purification procedures involving solvent extraction, thermal zone refining and nitric acid refiuxing are used in the current study. The effect of duration and type of solvent to dissolve impurities including fullerenes and P ACs (polyaromatic compounds) are monitored by nuclear magnetic reasonance, high performance liquid chromatography, and thermogravimetric analysis. Thermal zone refining yielded sample areas rich in nanotubes as seen by scanning electric microscopy. Refluxing in boiling nitric acid seem to improve the nanotube content. Different procedural steps are needed to purify samples produced by laser process compared to arc process. These alternative methods of nanotube purification will be presented along with results from supporting analytical techniques.

  1. Purification process for vertically aligned carbon nanofibers (United States)

    Nguyen, Cattien V.; Delziet, Lance; Matthews, Kristopher; Chen, Bin; Meyyappan, M.


    Individual, free-standing, vertically aligned multiwall carbon nanotubes or nanofibers are ideal for sensor and electrode applications. Our plasma-enhanced chemical vapor deposition techniques for producing free-standing and vertically aligned carbon nanofibers use catalyst particles at the tip of the fiber. Here we present a simple purification process for the removal of iron catalyst particles at the tip of vertically aligned carbon nanofibers derived by plasma-enhanced chemical vapor deposition. The first step involves thermal oxidation in air, at temperatures of 200-400 degrees C, resulting in the physical swelling of the iron particles from the formation of iron oxide. Subsequently, the complete removal of the iron oxide particles is achieved with diluted acid (12% HCl). The purification process appears to be very efficient at removing all of the iron catalyst particles. Electron microscopy images and Raman spectroscopy data indicate that the purification process does not damage the graphitic structure of the nanotubes.

  2. Antioxidant assay using genetically engineered bioluminescent Escherichia coli (United States)

    Bartolome, Amelita; Macalino, Bernadette; Pastoral, Ian Lemuel; Sevilla, Fortunato, III


    A new antioxidant activity assay based on the reactive oxygen species (ROS)-inducible bacterial strain (E. coli DPD2511) is described. The strain harbors the plasmid pKatG::luxCDABE and responds to hydrogen peroxide treatment by increasing light emission at 490 nm. Antioxidant capacity is evaluated through the ability of an agent to inhibit the hydrogen peroxide-induced bioluminescence of E. coli DPD2511. Applicability of the developed assay in detecting levels of antioxidants in various aqueous plant extracts is demonstrated. The assay was validated against 2,2-diphenylpicrylhydrazyl (DPPH) assay, a known antioxidant assay.

  3. Prevalence of Campylobacter in Dutch sewage purification plants

    NARCIS (Netherlands)

    Koenraad, P.M.F.J.


    Campylobacter bacteria are an important cause of bacterial gastro-enteritis in man. Although food of animal origin is the main source of human infection, a casecontrol study in the United States of America showed that 8% of all campylobacteriosis cases could be

  4. Prevalence of Campylobacter in Dutch sewage purification plants.

    NARCIS (Netherlands)

    Koenraad, P.M.F.J.


    Campylobacter bacteria are an important cause of bacterial gastro-enteritis in man. Although food of animal origin is the main source of human infection, a casecontrol study in the United States of America showed that 8% of all campylobacteriosis cases could be attributed to consumption of contamina

  5. Feeding preferences of Microtheca punctigera (Achard (Coleoptera: Chrysomelidae for some Brassicaceae plants in multiple-choice assays Preferência alimentar de Microtheca punctigera (Achard (Coleoptera: Chrysomelidae por algumas crucíferas em testes de mútipla escolha

    Directory of Open Access Journals (Sweden)

    Ayres Oliveira Menezes Jr.


    Full Text Available Host plant feeding preference is important basic information for the development of insect management strategies. Multiple-choice feeding preference assays were conducted in the laboratory for the chrysomelid beetle, Microtheca punctigera (Achard. Feeding was assessed 72 h after onset of experiments. With one larva per Petri dish, food items comprised watercress, Nasturtium officinale L., arugula, Eruca sativa L., mustard, Brassica juncea Cosson, Chinese cabbage, B. pekinensis (Lour. Rupr. and wild radish (Raphanus raphanistrum L.. Feeding ranking preferences were Chinese cabbage, mustard, wild radish, arugula and watercress (7.97, 1.85, 0.98, 0.36 and 0.11 mm², respectively. Feeding on Chinese cabbage was 4.31 times more intense than on mustard. The same experiment was repeated with one adult per dish. Responses of males and females were quite similar. Feeding was higher on mustard (87.2 and 142.8 for males and females, respectively. Feeding on arugula (51.5 and 132.7 and Chinese cabbage (51.8 and 89.0 were intermediate. Watercress (22.96 and 39.3 and wild radish (12.03 and 28.4 were the least preferred host plants. In a third experiment, ten larvae per dish were used and spinach, Tetragonia expansa Murr., radish, Raphanus sativus L. and collard, B. oleracea var. acephala L., were also included. Daily larval frequencies on each food were also measured. Feeding was similar on Chinese cabbage and mustard (47.89 and 53.78, respectively. Number of insects was greater on mustard, Chinese cabbage and wild radish. Probable explanations for results and proposals for further investigations are discussed.Preferência alimentar é informação básica importante para o desenvolvimento de estratégias de manejo. Experimentos de preferência alimentar com múltipla chance de escolha foram conduzidos em laboratório para o crisomelídeo Microtheca punctigera (Acherd. A alimentação foi avaliada 72h após o início dos experimentos. Com uma larva por placa

  6. Purification of the off-gases of the process of radioactive waste vitrification in induction melter

    Energy Technology Data Exchange (ETDEWEB)

    Gorbunov, V. A.; Katannikov, I. S.; Knyasev, O. A.; Kornev, V. I.; Lifanov, F. A.; Polkanov, M. A.; Savkin, A. E. [Moscow Scientific and Inndustrial Association RADON, Moscow (Russian Federation)


    Moscow SIA RADON has developed the method of vitrifying both radioactive ashes, arising from radioactive waste incineration, and liquid radioactive waste in induction melter. In the experimental plant the characteristics of off-gases were determined and various constructions of filters and filtering materials for dust trapping were tested. On the base of test results the plant for liquid radioactive waste vitrification has been constructed on the base of induction melter {sup c}old crucible{sup ,} equipped with modern effective dust and gas purification system, consisting of filtration unit, absorption unit and unit for nitrogen oxide catalytic reduction. (author). 3 refs., 9 tabs., 3 figs.

  7. Cell viability assays: introduction. (United States)

    Stoddart, Martin J


    The measurement of cell viability plays a fundamental role in all forms of cell culture. Sometimes it is the main purpose of the experiment, such as in toxicity assays. Alternatively, cell viability can be used to -correlate cell behaviour to cell number, providing a more accurate picture of, for example, anabolic -activity. There are wide arrays of cell viability methods which range from the most routine trypan blue dye exclusion assay to highly complex analysis of individual cells, such as using RAMAN microscopy. The cost, speed, and complexity of equipment required will all play a role in determining the assay used. This chapter aims to provide an overview of many of the assays available today.

  8. Tube-Forming Assays. (United States)

    Brown, Ryan M; Meah, Christopher J; Heath, Victoria L; Styles, Iain B; Bicknell, Roy


    Angiogenesis involves the generation of new blood vessels from the existing vasculature and is dependent on many growth factors and signaling events. In vivo angiogenesis is dynamic and complex, meaning assays are commonly utilized to explore specific targets for research into this area. Tube-forming assays offer an excellent overview of the molecular processes in angiogenesis. The Matrigel tube forming assay is a simple-to-implement but powerful tool for identifying biomolecules involved in angiogenesis. A detailed experimental protocol on the implementation of the assay is described in conjunction with an in-depth review of methods that can be applied to the analysis of the tube formation. In addition, an ImageJ plug-in is presented which allows automatic quantification of tube images reducing analysis times while removing user bias and subjectivity.

  9. Transgenic Animal Mutation Assays

    Institute of Scientific and Technical Information of China (English)

    Tao Chen; Ph.D.D.A.B.T.


    @@ The novel transgenic mouse and rat mutation assays have provided a tool for analyzing in vivo mutation in any tissue, thus permitting the direct comparison of cancer incidence with mutant frequency.

  10. Assays for thrombopoietin

    Energy Technology Data Exchange (ETDEWEB)

    McDonald, T.P.


    In summary, thrombopoietin levels have been determined indirectly by measuring thrombocytopoiesis in assay animals (platelet counting, measurement of isotope incorporation into newly formed platelets, changes in platelet sizes, or alterations in number and size of megakaryocytes) and by use of an immunoassay. Although much work remains, it seems clear at the present time that isotopic uptake into platelets of specially prepared assay mice (rebound-thrombocytosis) is superior to the other techniques now available for the measurement of thrombopoietin. However, the ideal assay for TSF which is specific, rapid, and inexpensive is yet to be developed. An immunoassay is in the development stage, but will require additional work before it can be utilized for the routine assay of TSF.

  11. New Rapid Spore Assay (United States)

    Kminek, Gerhard; Conley, Catharine


    The presentation will detail approved Planetary Protection specifications for the Rapid Spore Assay for spacecraft components and subsystems. Outlined will be the research and studies on which the specifications were based. The research, funded by ESA and NASA/JPL, was conducted over a period of two years and was followed by limited cleanroom studies to assess the feasibility of this assay during spacecraft assembly.

  12. EB technology for the purification of flue gases

    Energy Technology Data Exchange (ETDEWEB)

    Kojima, Takuji [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment


    Sulfur oxides and nitrogen oxides in flue gas from coal-combustion boilers in power plants, dioxins in flue gas from municipal waste incineration facilities and toxic volatile organic compounds (VOCs) in off-gas from painting or cleaning factories are among air pollutants for which emission is regulated by a law in Japan. Electron beam is the effective and easy controllable radiation source for treatment of these flue gases. This report describes outline of the results so far obtained at JAERI on electron beam treatment of flue gas. The removal performance higher than 90% at 10 kGy for flue gas containing 800 ppm SOx and 225 ppm NOx were achieved and being applied to real-scale power plants in Poland and China with expectation of cost reduction of 20% compared to conventional plants. Decomposition of dioxins in flue gas from solid waste incinerators is another project. Using an accelerator of 300 keV and 40 mA for treatment of real incineration gas at 200degC, we obtain 90% decomposition of dioxins at 15 kGy irradiation. Expansion of these flue gas purification technologies combined with low-energy electron accelerators is expected. (S. Ohno)

  13. Separation process design for isolation and purification of natural products

    DEFF Research Database (Denmark)

    Malwade, Chandrakant R.

    Natural products are defined as secondary metabolites produced by plants and form a vast pool of compounds with unlimited chemical and functional diversity. Many of these secondary metabolites are high value added chemicals that are frequently used as ingredients in food, cosmetics, pharmaceutica...... information is used to further optimize the process flow sheet. Chemometric methods have been used to extract molecular level information for understanding the process streams in relation to their separation operations....... and other consumer products. Therefore, process technology towards industrial scale production of such high value chemicals from plants has significant value. Natural products can be obtained in pure form via synthetic or semi-synthetic route, but due to their complicated nature these methods have not been...... developed to the extent of industrial production for majority of natural products. Thus, isolation and purification of such natural products from plants is the most viable way to obtain natural products in pure form. This PhD project is mainly concerned with the design of separation process to isolate...

  14. Intein-mediated purification system: mechanism and applications

    Institute of Scientific and Technical Information of China (English)

    Sarra setrerrahmane; Shuhua Tan


    The incorporation of self-cleaving protein elements into a variety of fusion-based purification systems; has been an important development in the area of recombinant protein purification. The self-cleaving capability of these tags has recently been combined with additional purification tags to generate novel and convenient protein purification methods. This review elucidates the properties of intein, the mechanism of the intein-based protein splicing and the progress of intein-based protein purification procedures, and recent advances in the applications of intein.

  15. Chemical looping integration with a carbon dioxide gas purification unit

    Energy Technology Data Exchange (ETDEWEB)

    Andrus, Jr., Herbert E.; Jukkola, Glen D.; Thibeault, Paul R.; Liljedahl, Gregory N.


    A chemical looping system that contains an oxidizer and a reducer is in fluid communication with a gas purification unit. The gas purification unit has at least one compressor, at least one dryer; and at least one distillation purification system; where the gas purification unit is operative to separate carbon dioxide from other contaminants present in the flue gas stream; and where the gas purification unit is operative to recycle the contaminants to the chemical looping system in the form of a vent gas that provides lift for reactants in the reducer.

  16. A robust robotic high-throughput antibody purification platform. (United States)

    Schmidt, Peter M; Abdo, Michael; Butcher, Rebecca E; Yap, Min-Yin; Scotney, Pierre D; Ramunno, Melanie L; Martin-Roussety, Genevieve; Owczarek, Catherine; Hardy, Matthew P; Chen, Chao-Guang; Fabri, Louis J


    Monoclonal antibodies (mAbs) have become the fastest growing segment in the drug market with annual sales of more than 40 billion US$ in 2013. The selection of lead candidate molecules involves the generation of large repertoires of antibodies from which to choose a final therapeutic candidate. Improvements in the ability to rapidly produce and purify many antibodies in sufficient quantities reduces the lead time for selection which ultimately impacts on the speed with which an antibody may transition through the research stage and into product development. Miniaturization and automation of chromatography using micro columns (RoboColumns(®) from Atoll GmbH) coupled to an automated liquid handling instrument (ALH; Freedom EVO(®) from Tecan) has been a successful approach to establish high throughput process development platforms. Recent advances in transient gene expression (TGE) using the high-titre Expi293F™ system have enabled recombinant mAb titres of greater than 500mg/L. These relatively high protein titres reduce the volume required to generate several milligrams of individual antibodies for initial biochemical and biological downstream assays, making TGE in the Expi293F™ system ideally suited to high throughput chromatography on an ALH. The present publication describes a novel platform for purifying Expi293F™-expressed recombinant mAbs directly from cell-free culture supernatant on a Perkin Elmer JANUS-VariSpan ALH equipped with a plate shuttle device. The purification platform allows automated 2-step purification (Protein A-desalting/size exclusion chromatography) of several hundred mAbs per week. The new robotic method can purify mAbs with high recovery (>90%) at sub-milligram level with yields of up to 2mg from 4mL of cell-free culture supernatant.

  17. A Heparin Purification Process Removes Spiked Transmissible Spongiform Encephalopathy Agent. (United States)

    Bett, Cyrus; Grgac, Ksenija; Long, Dianna; Karfunkle, Michael; Keire, David A; Asher, David M; Gregori, Luisa


    In 2000, bovine heparin was withdrawn from the US market for fear of contamination with bovine spongiform encephalopathy (BSE) agent, the cause of variant Creutzfeldt-Jakob disease in humans. Thus, US heparin is currently sourced only from pig intestines. Availability of alternative sources of crude heparin, a life-saving drug, would benefit public health. Bovine heparin is an obvious option, but BSE clearance by the bovine heparin manufacturing process should be evaluated. To this end, using hamster 263K scrapie as a surrogate for BSE agent, we applied a four-step bench-scale heparin purification protocol resembling a typical heparin manufacturing process to investigate removal of the spiked scrapie agent. We removed aliquots from each step and analyzed them for residual abnormal prion protein (PrP(TSE)) using a sensitive in vitro method, real-time quaking-induced conversion (RT-QuIC) assay, and for infectivity using animal bioassays. The purification process reduced infectivity by 3.6 log10 and removed PrP(TSE), measured as seeding activity, by 3.4 log10. NaOH treatment was the most effective removal step tested. We also investigated NaOH at different concentrations and pH: the results showed that as much as 5.2 log10 of PrP(TSE) seeding activity was removed at pH 12.5. Thus, changes to the concentration, treatment time, and temperature of alkaline extraction might further improve removal. Our results, using a basic heparin manufacturing process, inform efforts to reintroduce safe bovine heparin in the USA.

  18. FastTrack to supercritical fluid chromatographic purification: Implementation of a walk-up analytical supercritical fluid chromatography/mass spectrometry screening system in the medicinal chemistry laboratory. (United States)

    Aurigemma, Christine; Farrell, William


    Medicinal chemists often depend on analytical instrumentation for reaction monitoring and product confirmation at all stages of pharmaceutical discovery and development. To obtain pure compounds for biological assays, the removal of side products and final compounds through purification is often necessary. Prior to purification, chemists often utilize open-access analytical LC/MS instruments because mass confirmation is fast and reliable, and the chromatographic separation of most sample constituents is sufficient. Supercritical fluid chromatography (SFC) is often used as an orthogonal technique to HPLC or when isolation of the free base of a compound is desired. In laboratories where SFC is the predominant technique for analysis and purification of compounds, a reasonable approach for quickly determining suitable purification conditions is to screen the sample against different columns. This can be a bottleneck to the purification process. To commission SFC for open-access use, a walk-up analytical SFC/MS screening system was implemented in the medicinal chemistry laboratory. Each sample is automatically screened through six column/method conditions, and on-demand data processing occurs for the chromatographers after each screening method is complete. This paper highlights the "FastTrack" approach to expediting samples through purification.

  19. High-throughput immunoturbidimetric assays for in-process determination of polyclonal antibody concentration and functionality in crude samples

    DEFF Research Database (Denmark)

    Bak, Hanne; Kyhse-Andersen, J.; Thomas, O.R.T.


    We present fast, simple immunoturbidimetric assays suitable for direct determination of antibody 'concentration' and 'functionality' in crude samples, such as in-process samples taken at various stages during antibody purification. Both assays display excellent linearity and analytical recovery....... The 'functionality' assay displayed concentration dependent sensitivity to interference for ammonium sulphate and Tris(hydroxymethyl)-amino-methane, but was essentially unaffected by all other salts and buffer combinations tested. The immunoturbidimetric assays described here are generically applicable to polyclonal...... antibodies, require only basic laboratory equipment, are robust, fast, cheap, easy to perform, and readily adapted to automation....

  20. Partial purification and characterization of an endo-alpha-N-acetylgalactosaminidase from the culture medium of Streptomyces sp. OH-11242.


    Ishii-Karakasa, I; Iwase, H.; Hotta, K; Tanaka, Y.; Omura, S


    For the purification of a new type of endo-alpha-N-acetylgalactosaminidase from the culture medium of Streptomyces sp. OH-11242 (endo-GalNAc-ase-S) [Iwase, Ishii, Ishihara, Tanaka, Omura & Hotta (1988) Biochem. Biophys. Res. Commun. 151, 422-428], a method for assaying enzyme activity was established. Using purified pig gastric mucus glycoprotein (PGM) as the substrate, oligosaccharides liberated from PGM were pyridylaminated, and the reducing terminal sugars of oligosaccharides larger than G...

  1. Enhancing the water purification efficiency of a floating treatment wetland using a biofilm carrier. (United States)

    Zhang, Lingling; Zhao, Jing; Cui, Naxin; Dai, Yanran; Kong, Lingwei; Wu, Juan; Cheng, Shuiping


    Floating treatment wetlands (FTWs) and biofilm carriers are widely used in water purification. The objective of the present work was to explore whether and to what extent an FTW integrated with plants and biofilm carriers (FTW-I) could enhance the nutrient removal efficiency. Significantly higher removal rates of ammonia nitrogen (85.2 %), total phosphorus (82.7 %), and orthophosphate (82.5 %) were observed in the FTW-I treatment relative to the FTW with plants (FTW-P; 80.0, 78.5, and 77.6 %, respectively) and the FTW with biofilm carriers (FTW-B; 56.7, 12.9, and 13.4 %, respectively) (p < 0.05). The mass balance results indicated that plant uptake was the main pathway for N and P removal (accounting for 58.1 and 91.4 %, respectively) in FTW-I, in which only 1.2 % of the N and 5.7 % of the P was deposited on the bottom of the tank. In addition, the plants translocated 43.9 and 80.2 % of the N and P in the water and 83.5 and 88.3 % of the absorbed N and P, respectively, into their aboveground tissues. The combination of an FTW and biofilm carriers can improve the efficiency of water purification, and nutrients can be rapidly removed from the system by harvesting the aboveground plant tissues.

  2. Purification of phosphinothricin acetyltransferase using Reactive brown 10 affinity in a single chromatography step. (United States)

    Wang, Cunxi; Lee, Thomas C; Crowley, Kathleen S; Bell, Erin


    The expression of phosphinothricin N-acetyltransferase (PAT) protein in transgenic plants confers tolerance to the herbicide glufosinate. To enable the characterization of PAT protein expressed in plants, it is necessary to obtain high purity PAT protein from the transgenic grain. Because transgenically expressed proteins are typical present at very low levels (i.e. 0.1-50 μg protein/g grain), a highly specific and efficient purification protocol is required to purify them. Based on the physicochemical properties of PAT, we developed a novel purification method that is simple, time-saving, inexpensive and reproducible. The novel method employs a single chromatography step using a reactive dye resin, Reactive brown 10-agarose. Reactive brown 10 preferentially binds the PAT protein, which can then be specifically released by one of its substrates, acetyl-CoA. Using Reactive brown 10-agarose, PAT protein was purified to homogeneity from cottonseed with high recovery efficiency. As expected, the Reactive brown 10-produced PAT was enzymatically active. Other applications of the method on protein expression and purification, and development of PAT enzymatic inhibitors were also discussed. Copyright © 2013 Elsevier Inc. All rights reserved.

  3. Soils and waste water purification from oil products using combined methods under the North conditions. (United States)

    Evdokimova, Galina A; Gershenkop, Alexander Sh; Mozgova, Natalia P; Myazin, Vladimir A; Fokina, Nadejda V


    Oil and gas production and transportation in Russia is increasingly moving to the north regions. Such regions are characterized by relatively low self-purification capacity of the natural environments from the contaminants due to slow character of the energy exchange and mass transfer processes. Off-shore field development in the Barents Sea and oil product transportation can result in contamination, as confirmed by the national and international practice of the developed oil and gas regions. The research aims at development of the soil bioremediation methods and industrial waste water purification contaminated by oil products in the north-western region of Russia. The dynamics of oil products carry-over have been investigated under the field model experiments in podzolic soils: gas condensate, diesel fuel and mazut from oil and the plants were selected for phyto-remediation of contaminated soils under high north latitudes. It is shown that soil purification from light hydrocarbons takes place during one vegetation period. In three months of the vegetation period the gas condensate was completely removed from the soil, diesel fuel - almost completely (more than 90%). Residual amounts of heavy hydrocarbons were traced, even 1.5 later. The following plants that were highly resistant to the oil product contamination were recommended for bioremediation: Phalaroides arundinacea, Festuca pratensis, Phleum pratense, Leymus arenarius. There has been developed and patented the combined method of treatment of waste water contaminated with hydrocarbons based on inorganic coagulants and local oil-oxidizing bacteria.

  4. Purification of functionalized DNA origami nanostructures. (United States)

    Shaw, Alan; Benson, Erik; Högberg, Björn


    The high programmability of DNA origami has provided tools for precise manipulation of matter at the nanoscale. This manipulation of matter opens up the possibility to arrange functional elements for a diverse range of applications that utilize the nanometer precision provided by these structures. However, the realization of functionalized DNA origami still suffers from imperfect production methods, in particular in the purification step, where excess material is separated from the desired functionalized DNA origami. In this article we demonstrate and optimize two purification methods that have not previously been applied to DNA origami. In addition, we provide a systematic study comparing the purification efficacy of these and five other commonly used purification methods. Three types of functionalized DNA origami were used as model systems in this study. DNA origami was patterned with either small molecules, antibodies, or larger proteins. With the results of our work we aim to provide a guideline in quality fabrication of various types of functionalized DNA origami and to provide a route for scalable production of these promising tools.

  5. Purification of His-Tagged Proteins. (United States)

    Spriestersbach, Anne; Kubicek, Jan; Schäfer, Frank; Block, Helena; Maertens, Barbara


    Ni-NTA affinity purification of His-tagged proteins is a bind-wash-elute procedure that can be performed under native or denaturing conditions. Here, protocols for purification of His-tagged proteins under native, as well as under denaturing conditions, are given. The choice whether to purify the target protein under native or denaturing conditions depends on protein location and solubility, the accessibility of the His tag, and the desired downstream application. His-tagged proteins can be purified by a single-step affinity chromatography, namely immobilized metal ion affinity chromatography (IMAC), which is commercially available in different kinds of formats, Ni-NTA matrices being the most widely used. The provided protocols describe protein purification in the batch binding mode and apply gravity-assisted flow in disposable columns; this procedure is simple to conduct and extremely robust. IMAC purification can equally be performed in prepacked columns using FPLC or other liquid chromatography instrumentation, or using magnetic bead-based methods (Block et al., 2009).

  6. Simple and Economic Purification Method of Berberine


    SHU-HUA, LU; Xu, Li; GUI-BAO, LU; TOYOKAZU, KISHI; Setsuko, SEKITA; Motoyoshi, SATAKE; Tianjin Institute for Drug Control; Japan International Corporation Agency; National Institute of Health Sciences


    The authers examined for the purification and quantitative analysis of berberine, thin layer chromatography, centrifugal round plate thin layer chromatography, recrystallization, dry silica gel column chromatography, wet silica gel column chromatography, solvent partition and preparative thin layer chromatography. We established a simple and economic method which is a combination of solvent partition and preparative thin layer chromatography.

  7. Expression and Purification of Sperm Whale Myoglobin (United States)

    Miller, Stephen; Indivero, Virginia; Burkhard, Caroline


    We present a multiweek laboratory exercise that exposes students to the fundamental techniques of bacterial expression and protein purification through the preparation of sperm whale myoglobin. Myoglobin, a robust oxygen-binding protein, contains a single heme that gives the protein a reddish color, making it an ideal subject for the teaching…

  8. Enterovirus 71 Virus Propagation and Purification


    Kristin L Shingler; Organtini, Lindsey J.; Hafenstein, Susan


    Since its discovery in 1969, enterovirus 71 (EV71) has emerged as a serious worldwide health threat. This member of the picornavirus family causes hand, foot, and mouth disease, and also has the capacity to invade the central nervous system to cause severe disease and death. This is the propagation and purification procedure to produce infectious virion.

  9. Expression and Purification of Sperm Whale Myoglobin (United States)

    Miller, Stephen; Indivero, Virginia; Burkhard, Caroline


    We present a multiweek laboratory exercise that exposes students to the fundamental techniques of bacterial expression and protein purification through the preparation of sperm whale myoglobin. Myoglobin, a robust oxygen-binding protein, contains a single heme that gives the protein a reddish color, making it an ideal subject for the teaching…

  10. Evaluation of immunoglobulin purification methods and their impact on quality and yield of antigen-specific antibodies

    Directory of Open Access Journals (Sweden)

    Khan Farhat


    Full Text Available Abstract Background Antibodies are the main effectors against malaria blood-stage parasites. Evaluation of functional activities in immune sera from Phase 2a/b vaccine trials may provide invaluable information in the search for immune correlates of protection. However, the presence of anti-malarial-drugs, improper collection/storage conditions or concomitant immune responses against other pathogens can contribute to non-specific anti-parasite activities when the sera/plasma are tested in vitro. Purification of immunoglobulin is a standard approach for reducing such non-specific background activities, but the purification method itself can alter the quality and yield of recovered Ag-specific antibodies. Methods To address this concern, various immunoglobulin (Ig purification methods (protein G Sepharose, protein A/G Sepharose, polyethylene glycol and caprylic acid-ammonium sulphate precipitation were evaluated for their impact on the quality, quantity and functional activity of purified rabbit and human Igs. The recovered Igs were analysed for yield and purity by SDS-PAGE, for quality by Ag-specific ELISAs (determining changes in titer, avidity and isotype distribution and for functional activity by in vitro parasite growth inhibition assay (GIA. Results This comparison demonstrated that overall polyethylene glycol purification of human serum/plasma samples and protein G Sepharose purification of rabbit sera are optimal for recovering functional Ag-specific antibodies. Conclusion Consequently, critical consideration of the purification method is required to avoid selecting non-representative populations of recovered Ig, which could influence interpretations of vaccine efficacy, or affect the search for immune correlates of protection.

  11. Computer-Aided Modelling of Short-Path Evaporation for Chemical Product Purification, Analysis and Design

    DEFF Research Database (Denmark)

    Sales-Cruz, Alfonso Mauricio; Gani, Rafiqul


    An important stage in the design process for many chemical products is its manufacture where, for a class of chemical products that may be thermally unstable (such as, drugs, insecticides, flavours /fragrances, and so on), the purification step plays a major role. Short-path evaporation is a safe...... method, suitable for separation and purification of thermally unstable materials whose design and analysis can be efficiently performed through reliable model-based techniques. This paper presents a generalized model for short-path evaporation and highlights its development, implementation and solution...... glycerol, mono-, di- and triglycerides, and (b) the recovery of a pharmaceutical product from a six-component mixture. Validation of the short-path evaporation model is highlighted through the comparison of experimental data from an industrial pilot plant with the simulated results from the model. Also...



    Rakhmatulin I.R.


    The article discusses the possibility of using renewable energy for water purification. Results of analysis of a preferred energy source for a water purification using installed in places where fresh water shortages and a lack of electrical energy. The possibility of desalination of salt water using solar energy for regions with temperate climate. Presented desalination plant working on energy vacuum solar collectors, principles of action developed by the desalination plant. The experimental ...

  13. Against vaccine assay secrecy. (United States)

    Herder, Matthew; Hatchette, Todd F; Halperin, Scott A; Langley, Joanne M


    Increasing the transparency of the evidence base behind health interventions such as pharmaceuticals, biologics, and medical devices, has become a major point of critique, conflict, and policy focus in recent years. Yet the lack of publicly available information regarding the immunogenicity assays upon which many important, widely used vaccines are based has received no attention to date. In this paper we draw attention to this critical public health problem by reporting on our efforts to secure vaccine assay information in respect of 10 vaccines through Canada's access to information law. We argue, under Canadian law, that the public health interest in having access to the methods for these laboratory procedures should override claims by vaccine manufacturers and regulators that this information is proprietary; and, we call upon several actors to take steps to ensure greater transparency with respect to vaccine assays, including regulators, private firms, researchers, research institutions, research funders, and journal editors.

  14. Against vaccine assay secrecy (United States)

    Herder, Matthew; Hatchette, Todd F; Halperin, Scott A; Langley, Joanne M


    Increasing the transparency of the evidence base behind health interventions such as pharmaceuticals, biologics, and medical devices, has become a major point of critique, conflict, and policy focus in recent years. Yet the lack of publicly available information regarding the immunogenicity assays upon which many important, widely used vaccines are based has received no attention to date. In this paper we draw attention to this critical public health problem by reporting on our efforts to secure vaccine assay information in respect of 10 vaccines through Canada's access to information law. We argue, under Canadian law, that the public health interest in having access to the methods for these laboratory procedures should override claims by vaccine manufacturers and regulators that this information is proprietary; and, we call upon several actors to take steps to ensure greater transparency with respect to vaccine assays, including regulators, private firms, researchers, research institutions, research funders, and journal editors. PMID:25826194

  15. Development of an mlrA Gene-Directed TaqMan PCR Assay for Quantitative Assessment of Microcystin-Degrading Bacteria within Water Treatment Plant Sand Filter Biofilms▿ (United States)

    Hoefel, Daniel; Adriansen, Caroline M. M.; Bouyssou, Magali A. C.; Saint, Christopher P.; Newcombe, Gayle; Ho, Lionel


    We report for the first time a quantitative mlrA gene-directed TaqMan PCR assay for the rapid detection of microcystin-degrading bacteria. This was applied, in combination with 16S ribosomal DNA-directed quantitative PCR and denaturing gradient gel electrophoresis, to study virgin sand filter column biofilm development and to correlate mlrA gene abundance with microcystin removal efficiency. PMID:19502429

  16. Secretory leukocyte protease inhibitor (SLPI) might contaminate murine monoclonal antibodies after purification on protein G. (United States)

    Schenk, Jörg A; Fettke, Joerg; Lenz, Christine; Albers, Katharina; Mallwitz, Frank; Gajovic-Eichelmann, Nenad; Ehrentreich-Förster, Eva; Kusch, Emely; Sellrie, Frank


    The large scale production of a monoclonal anti-progesterone antibody in serum free medium followed by affinity chromatography on protein G lead to a contamination of the antibody sample with a protein of about 14 kDa. This protein was identified by mass spectrometry as secretory leukocyte protease inhibitor (SLPI). This SLPI contamination lead to a failure of the fiber-optic based competitive fluorescence assay to detect progesterone in milk. Purification of the monoclonal antibody using protein A columns circumvented this problem.

  17. Expression and purification of soluble porcine cystatin 11 in Pichia pastoris. (United States)

    Fan, Kuohai; Jiang, Junbing; Wang, Zhirui; Fan, Ruicheng; Yin, Wei; Sun, Yaogui; Li, Hongquan


    Cystatin 11 (CST11) belongs to the cystatin type 2 family of cysteine protease inhibitors and exhibits antimicrobial activity in vitro. In this study, we describe the expression and purification of recombinant porcine CST11 in the Pichia pastoris system. We then assess its antimicrobial activity against Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus subtilis by liquid growth inhibition assay. Kinetic studies indicate that the recombinant porcine CST11 has high potency against E. coli and S. aureus. Scanning electronic microscope analysis showed that CST11 might be targeting the bacterial membrane and, thus, could potentially be developed as a therapeutic agent for inhibiting microbe infection without the risk of antibiotic resistance.

  18. Rapid Purification of Salmonella DNA in Minced Meat and Detection by Real-time PCR

    DEFF Research Database (Denmark)

    Jenikova, G.; Jensen, Annette Nygaard; Demnerova, K.;


    of DNeasy was found to be 6-8 CFU in just 19 end-point fluorescence (C-t) values, while this was 22 C-t for a combination of DNeasy and BactXtractor. Extraction by DNeasy resulted in C-t ...Four rapid and simple DNA purification and sample treatment protocols were evaluated for detection of Salmonella enterica in spiked minced meat, using a fluorogenic 5' nuclease (TaqMan) PCR assay in an ABI-Prism 7700 Sequence Detector. The detection limit with the single separation treatment...

  19. CTL ELISPOT assay. (United States)

    Ranieri, Elena; Popescu, Iulia; Gigante, Margherita


    Enzyme-linked immune absorbent spot (Elispot) is a quantitative method for measuring relevant parameters of T cell activation. The sensitivity of Elispot allows the detection of low-frequency antigen-specific T cells that secrete cytokines and effector molecules, such as granzyme B and perforin. Cytotoxic T cell (CTL) studies have taken advantage with this high-throughput technology by providing insights into quantity and immune kinetics. Accuracy, sensitivity, reproducibility, and robustness of Elispot resulted in a wide range of applications in research as well as in diagnostic field. Actually, CTL monitoring by Elispot is a gold standard for the evaluation of antigen-specific T cell immunity in clinical trials and vaccine candidates where the ability to detect rare antigen-specific T cells is of relevance for immune diagnostic. The most utilized Elispot assay is the interferon-gamma (IFN-γ) test, a marker for CD8(+) CTL activation, but Elispot can also be used to distinguish different subsets of activated T cells by using other cytokines such as T-helper (Th) 1-type cells (characterized by the production of IFN-γ, IL-2, IL-6, IL-12, IL-21, and TNF-α), Th2 (producing cytokines like IL-4, IL-5, IL-10, and IL-13), and Th17 (IL-17) cells. The reliability of Elispot-generated data, by the evaluation of T cell frequency recognizing individual antigen/peptide, is the core of this method currently applied widely to investigate specific immune responses in cancer, infections, allergies, and autoimmune diseases. The Elispot assay is competing with other methods measuring single-cell cytokine production, e.g., intracellular cytokine by FACS or Miltenyi cytokine secretion assay. Other types of lymphocyte frequency and function assays include limiting dilution assay (LDA), cytotoxic T cell assay (CTL), and tetramer staining. Compared with respect to sensitivity the Elispot assay is outranking other methods to define frequency of antigen-specific lymphocytes. The method

  20. Assays for calcitonin receptors

    Energy Technology Data Exchange (ETDEWEB)

    Teitelbaum, A.P.; Nissenson, R.A.; Arnaud, C.D.


    The assays for calcitonin receptors described focus on their use in the study of the well-established target organs for calcitonin, bone and kidney. The radioligand used in virtually all calcitonin binding studies is /sup 125/I-labelled salmon calcitonin. The lack of methionine residues in this peptide permits the use of chloramine-T for the iodination reaction. Binding assays are described for intact bone, skeletal plasma membranes, renal plasma membranes, and primary kidney cell cultures of rats. Studies on calcitonin metabolism in laboratory animals and regulation of calcitonin receptors are reviewed.

  1. Purification and characterization of broccoli (Brassica oleracea var. italica) myrosinase (β-thioglucosidase glucohydrolase). (United States)

    Mahn, Andrea; Angulo, Alejandro; Cabañas, Fernanda


    Myrosinase (β-thioglucosidase glucohydrolase, EC from broccoli (Brassica oleracea var. italica) was purified by ammonium sulfate precipitation followed by concanavalin A affinity chromatography, with an intermediate dialysis step, resulting in 88% recovery and 1318-fold purification. These are the highest values reported for the purification of any myrosinase. The subunits of broccoli myrosinase have a molecular mass of 50-55 kDa. The native molecular mass of myrosinase was 157 kDa, and accordingly, it is composed of three subunits. The maximum activity was observed at 40 °C and at pH below 5.0. Kinetic assays demonstrated that broccoli myrosinase is subjected to substrate (sinigrin) inhibition. The Michaelis-Menten model, considering substrate inhibition, gave Vmax equal to 0.246 μmol min(-1), Km equal to 0.086 mM, and K(I) equal to 0.368 mM. This is the first study about purification and characterization of broccoli myrosinase.

  2. Recombinant expression and purification of an ATP-dependent DNA ligase from Aliivibrio salmonicida. (United States)

    Williamson, Adele; Pedersen, Hege


    The genome of the psychrophilic fish-pathogen Aliivibrio salmonicida encodes a putative ATP-dependent DNA ligase in addition to a housekeeping NAD-dependent enzyme. In order to study the structure and activity of the ATP dependent ligase in vitro we have undertaken its recombinant production and purification from an Escherichia coli based expression system. Expression and purification of this protein presented two significant challenges. First, the gene product was moderately toxic to E. coli cells, second it was necessary to remove the large amounts of E. coli DNA present in bacterial lysates without contamination of the protein preparation by nucleases which might interfere with future assaying. The toxicity problem was overcome by fusion of the putative ligase to large solubility tags such as maltose-binding protein (MBP) or Glutathione-S-transferase (GST), and DNA was removed by treatment with a nuclease which could be inhibited by reducing agents. As the A. salmonicida ATP-dependent DNA ligase gene encodes a predicted leader peptide, both the full-length and mature forms of the protein were produced. Both possessed ATP-dependent DNA ligase activity, but the truncated form was significantly more active. Here we detail the first reported production, purification and preliminary characterization of active A. salmonicida ATP-dependent DNA ligase.

  3. Transient expression assays in tobacco protoplasts. (United States)

    Vanden Bossche, Robin; Demedts, Brecht; Vanderhaeghen, Rudy; Goossens, Alain


    The sequence information generated through genome and transcriptome analysis from plant tissues has reached unprecedented sizes. Sequence homology-based annotations may provide hints for the possible function and roles of particular plant genes, but the functional annotation remains nonexistent or incomplete for many of them. To discover gene functions, transient expression assays are a valuable tool because they can be done more rapidly and at a higher scale than generating stably transformed tissues. Here, we describe a transient expression assay in protoplasts derived from suspension cells of tobacco (Nicotiana tabacum) for the study of the transactivation capacities of transcription factors. To enhance throughput and reproducibility, this method can be automated, allowing medium-throughput screening of interactions between large compendia of potential transcription factors and gene promoters.

  4. In Vitro Antibacterial, Antifungal, Antibiofilm, Antioxidant, and Anticancer Properties of Isosteviol Isolated from Endangered Medicinal Plant Pittosporum tetraspermum

    Directory of Open Access Journals (Sweden)

    Naif Abdullah Al-Dhabi


    Full Text Available This study aimed to investigate the in vitro antibacterial, antifungal, antibiofilm, antioxidant, and anticancer properties of isosteviol isolated from endangered medicinal plant Pittosporum tetraspermum. Pure compound was obtained and characterized by column chromatography followed by 1H NMR, 13C NMR, IR, and mass spectral analysis. The antimicrobial activities of the compound were assessed by the broth microdilution method and the antioxidant properties were determined using reducing ability assay, DPPH scavenging assay, hydroxyl radical scavenging activity, and superoxide radical scavenging assay. Anticancer study was evaluated by following MTT assay. Column purification and spectrocopical analysis lead to identifying isosteviol from the crude ethyl acetate extract. The compound exhibited significant activity against bacteria such as Staphylococcus epidermidis (125 µg/mL, Staphylococcus aureus (125 µg/mL, and Klebsiella pneumoniae (62.5 µg/mL. The MIC of the compound against Candida albicans, Aspergillus niger, and Trichophyton mentagrophytes was 62.5, 125, and 500 µg/mL, respectively. The compound showed comparatively better antibiofilm activity against E. coli, S. typhi, and P. aeruginosa. Furthermore, it exhibited good antioxidant properties. Anticancer properties of the compound against Vero and MCF7 cell lines were its advantage. Novel isosteviol would be useful to reduce the infectious diseases caused by pathogenic microorganisms or slow the progress of various oxidative stress-related diseases.

  5. Comet assay on tetraploid yeast cells

    DEFF Research Database (Denmark)

    Rank, Jette; Syberg, Kristian; Jensen, Klara


    Tetraploid yeast cells (Saccharomyces cerevisiae) were used in the comet assay with the intention of developing a new, fast and easy assay for detecting environmental genotoxic agents without using higher organisms. Two DNA-damaging chemicals, H2O2 and acrylamide, together with wastewater from...... three municipal treatment plants were tested for their effect on the yeast-cell DNA. The main problem with using yeast in the comet assay is the necessity to degrade the cell wall. This was achieved by using Zymolase 100 T twice during the procedure, since Zymolase 20 T did not open the cell wall....... Analytical problems that arose due to the small amount of DNA in the yeast nuclei in haploid and diploid cells, which contain 13 Mbp and 26 Mbp DNA per cell, respectively, were solved by using tetraploid yeast cells (52 Mbp) instead. DNA damage was shown after exposure to H2O2 and acrylamide. The lowest dose...

  6. New oligosaccharyltransferase assay method. (United States)

    Kohda, Daisuke; Yamada, Masaki; Igura, Mayumi; Kamishikiryo, Jun; Maenaka, Katsumi


    We developed a new in vitro assay for oligosaccharyltransferase (OST), which catalyzes the transfer of preassembled oligosaccharides on lipid carriers onto asparagine residues in polypeptide chains. The asparagine residues reside in the sequon, Asn-X-Thr/Ser, where X can be any amino acid residue except Pro. We demonstrate the potency of our assay using the OST from yeast. In our method, polyacrylamide gel electrophoresis is used to separate the glycopeptide products from the peptide substrates. The substrate peptide is fluorescently labeled and the formation of glycopeptides is analyzed by fluorescence gel imaging. Two in vitro OST assay methods are now widely used, but both the methods depend on previous knowledge of the oligosaccharide moiety: One method uses lectin binding as the separation mechanism and the other method uses biosynthetically or chemoenzymatically synthesized lipid-linked oligosaccharides as donors. N-linked protein glycosylation is found in all three domains of life, but little is known about the N-glycosylation in Archaea. Thus, our new assay, which does not require a priori knowledge of the oligosaccharides, will be useful in such cases. Indeed, we have detected the OST activity in the membrane fraction from a hyperthermophilic archaeon, Pyrococcus furiosus.

  7. Hyaluronic Acid Assays

    DEFF Research Database (Denmark)

    Itenov, Theis S; Kirkby, Nikolai S; Bestle, Morten H


    BACKGROUD: Hyaluronic acid (HA) is proposed as a marker of functional liver capacity. The aim of the present study was to compare a new turbidimetric assay for measuring HA with the current standard method. METHODS: HA was measured by a particle-enhanced turbidimetric immunoassay (PETIA) and enzyme...

  8. Instrument for assaying radiation

    Energy Technology Data Exchange (ETDEWEB)

    Coleman, Jody Rustyn; Farfan, Eduardo B.


    An instrument for assaying radiation includes a flat panel detector having a first side opposed to a second side. A collimated aperture covers at least a portion of the first side of the flat panel detector. At least one of a display screen or a radiation shield may cover at least a portion of the second side of the flat panel detector.

  9. Effect of Different Purification Techniques on the Characteristics of Heteropolysaccharide-Protein Biopolymer from Durian (Durio zibethinus Seed

    Directory of Open Access Journals (Sweden)

    Hamed Mirhosseini


    Full Text Available Natural biopolymers from plant sources contain many impurities (e.g., fat, protein, fiber, natural pigment and endogenous enzymes, therefore, an efficient purification process is recommended to minimize these impurities and consequently improve the functional properties of the biopolymer. The main objective of the present study was to investigate the effect of different purification techniques on the yield, protein content, solubility, water- and oil-holding capacity of a heteropolysaccharide-protein biopolymer obtained from durian seed. Four different purification methods using different chemicals and solvents (i.e., A (isopropanol and ethanol, B (isopropanol and acetone, C (saturated barium hydroxide, and D (Fehling solution] to liberate the purified biopolymer from its crude form were compared. In most cases, the purification process significantly (p < 0.05 improved the physicochemical properties of heteropolysaccharide-protein biopolymer from durian fruit seed. The present work showed that the precipitation using isopropanol and acetone (Method B resulted in the highest purification yield among all the tested purification techniques. The precipitation using saturated barium hydroxide (Method C led to induce the highest solubility and relatively high capacity of water absorption. The current study reveals that the precipitation using Fehling solution (Method D most efficiently eliminates the protein fraction, thus providing more pure biopolymer suitable for biological applications.


    Institute of Scientific and Technical Information of China (English)

    李文芬; 刘沛芬; 颜亨梅; 付秀芹; 汪波; 许华


    javanica〉Canna indica 〉Scirpus validus Vahl, whereas the order for root hair growth was Phragmites communis Trirn 〉 Scirpus validus Vahl 〉Oenanthe javanica 〉Canna indica 〉 Eichhornia crassi pe. Significant differences in absorption of COD, NHa-N, TN, TP were found among these five species. Rate of COD removal per unit area was ranked in the following order: Canna indica 〉Eichhornia crassipe〉Phragmites communis Trirn〉Scirpus validus Vahl〉Oenanthe javanica. NH3-N removal rate per unit area was Canna indica Phragrnites communis Trirn 〉 Eichhornia crassipe 〉 Scirpus validus Vahl 〉 Oenanthe javanica. Nitrogen absorption per unit area was Canna indica 〉 Eichhornia crassipe 〉 Phragrnites communis Trirn 〉 Oenanthe javanica〉Scirpus validus Vahl. Nitrogen and phosphate uptake rates showed the same rank order. It was determined that for creek water purification, canna can be used as the major plant, with reed as a supplement, together with small portions of water onion and water celery.

  11. Identification of protein interacting partners using tandem affinity purification. (United States)

    Bailey, Dalan; Urena, Luis; Thorne, Lucy; Goodfellow, Ian


    A critical and often limiting step in understanding the function of host and viral proteins is the identification of interacting cellular or viral protein partners. There are many approaches that allow the identification of interacting partners, including the yeast two hybrid system, as well as pull down assays using recombinant proteins and immunoprecipitation of endogenous proteins followed by mass spectrometry identification(1). Recent studies have highlighted the utility of double-affinity tag mediated purification, coupled with two specific elution steps in the identification of interacting proteins. This approach, termed Tandem Affinity Purification (TAP), was initially used in yeast(2,3) but more recently has been adapted to use in mammalian cells(4-8). As proof-of-concept we have established a tandem affinity purification (TAP) method using the well-characterized eukaryotic translation initiation factor eIF4E(9,10).The cellular translation factor eIF4E is a critical component of the cellular eIF4F complex involved in cap-dependent translation initiation(10). The TAP tag used in the current study is composed of two Protein G units and a streptavidin binding peptide separated by a Tobacco Etch Virus (TEV) protease cleavage sequence. The TAP tag used in the current study is composed of two Protein G units and a streptavidin binding peptide separated by a Tobacco Etch Virus (TEV) protease cleavage sequence(8). To forgo the need for the generation of clonal cell lines, we developed a rapid system that relies on the expression of the TAP-tagged bait protein from an episomally maintained plasmid based on pMEP4 (Invitrogen). Expression of tagged murine eIF4E from this plasmid was controlled using the cadmium chloride inducible metallothionein promoter. Lysis of the expressing cells and subsequent affinity purification via binding to rabbit IgG agarose, TEV protease cleavage, binding to streptavidin linked agarose and subsequent biotin elution identified numerous

  12. Purification and characterization of tyrosinase from walnut leaves (Juglans regia). (United States)

    Zekiri, Florime; Molitor, Christian; Mauracher, Stephan G; Michael, Claudia; Mayer, Rupert L; Gerner, Christopher; Rompel, Annette


    Polyphenol oxidase (PPO) is a type-3 copper enzyme catalyzing the oxidation of phenolic compounds to their quinone derivates, which are further converted to melanin, a ubiquitous pigment in living organisms. In this study a plant originated tyrosinase was isolated from walnut leaves (Juglans regia) and biochemically characterized. It was possible to isolate and purify the enzyme by means of an aqueous two-phase extraction method followed by chromatographic purification and identification. Interestingly, the enzyme showed a rather high monophenolase activity considering that the main part of plant PPOs with some exceptions solely possess diphenolase activity. The average molecular mass of 39,047 Da (Asp(101)→Arg(445)) was determined very accurately by high resolution mass spectrometry. This proteolytically activated tyrosinase species was identified as a polyphenol oxidase corresponding to the known jrPPO1 sequence by peptide sequencing applying nanoUHPLC-ESI-MS/MS. The polypeptide backbone with sequence coverage of 96% was determined to start from Asp(101) and not to exceed Arg(445).

  13. High-throughput process development of purification alternatives for the protein avidin. (United States)

    Diederich, Patrick; Hoffmann, Marc; Hubbuch, Jürgen


    With an increased number of applications in the field of the avidin-biotin technology, the resulting demand for highly-purified protein avidin has drawn our attention to the purification process of avidin that naturally occurs in chicken egg white. The high-throughput process development (HTPD) methodology was exploited, in order to evaluate purification process alternatives to commonly used ion-exchange chromatography. In a high-throughput format, process parameters for aqueous two-phase extraction, selective precipitation with salts and polyethylene glycol, and hydrophobic interaction and mixed-mode column chromatography experiments were performed. The HTPD strategy was complemented by a high-throughput tandem high-performance liquid chromatography assay for protein quantification. Suitable conditions for the separation of avidin from the major impurities ovalbumin, ovomucoid, ovotransferrin, and lysozyme were identified in the screening experiments. By combination of polyethylene glycol precipitation with subsequent resolubilization and separation in a polyethylene glycol/sulfate/sodium chloride two-phase system an avidin purity of 77% was obtained with a yield >90% while at the same time achieving a significant reduction of the process volume. The two-phase extraction and precipitation results were largely confirmed in larger scale with scale-up factors of 230 and 133, respectively. Seamless processing of the avidin enriched bottom phase was found feasible by using mixed-mode chromatography. By gradient elution a final avidin purity of at least 97% and yield >90% was obtained in the elution pool. The presented identification of a new and beneficial alternative for the purification of the high value protein thus represents a successful implementation of HTPD for an industrially relevant purification task. © 2015 American Institute of Chemical Engineers.

  14. Plant Comet Assay and Its Application on Environment Pollution Monitoring%植物彗星实验及其在生态毒理监测中的应用

    Institute of Scientific and Technical Information of China (English)

    曹玉伟; 马军; 郭长虹; 李蕊; 余建平


    植物彗星实(Comet assay)是近几年发展起来的一项检测DNA损伤的新技术,具有快速、简便、灵敏性高和可靠性强等特点.本文总结了近年来国内外植物彗星实验的研究成果,详细介绍了植物彗星实验方法及其在生态毒理学中的应用.

  15. Purification and characterization of M-177, a 177 kDa allergen, from the house dust mite Dermatophagoides farinae

    Directory of Open Access Journals (Sweden)

    Akihiro Fujikawa


    Full Text Available A high molecular weight allergen, M-177, was recently discovered in the house dust mite, Dermatophagoides farinae (D. farinae. The aims of this study were to develop a conventional purification procedure for M-177 and then to analyze some of the immunochemical properties of M-177. Mite extracts obtained from purified mite bodies were a suitable material for preparing M-177, because the purified mite extract contained large amounts of M-177. The purification of this allergen from the extract was achieved by ethanol fractionation, anion exchange chromatography, and gel filtration chromatography. The purified antigen was immunochemically equivalent to that of a preparation obtained by a previous affinity method using an anti-Mag 3-immobilized column. The yield of this purification procedure was 36.8% of the initial amount of M-177 in the extract, 40-fold greater than that of the previous immunoaffinity method. Our purification method was useful for preparing this allergen. The purified M-177 reacted in skin tests in 11 of 16 miteallergic patients, compared to 10 of 16 with Der f 2. The amount of M-177 in the purified mite extract determined by enzyme-linked immunosorbent assay inhibition was as much as 0.95% of the total protein, which was higher than the amounts of Der f 1 (0.52% and Der f 2 (0.32%. The potent allergenic activity and large amount of M-177 in the mites indicate that it is an important mite allergen.

  16. Purification of crude biodiesel using dry washing and membrane technologies

    Directory of Open Access Journals (Sweden)

    I.M. Atadashi


    Full Text Available Purification of crude biodiesel is mandatory for the fuel to meet the strict international standard specifications for biodiesel. Therefore, this paper carefully analyzed recently published literatures which deal with the purification of biodiesel. As such, dry washing technologies and the most recent membrane biodiesel purification process have been thoroughly examined. Although purification of biodiesel using dry washing process involving magnesol and ion exchange resins provides high-quality biodiesel fuel, considerable amount of spent absorbents is recorded, besides the skeletal knowledge on its operating process. Further, recent findings have shown that biodiesel purification using membrane technique could offer high-quality biodiesel fuel with less wastewater discharges. Thus, both researchers and industries are expected to benefit from the development of membrane technique in purifying crude biodiesel. As well biodiesel purification via membranes has been shown to be environmentally friendly. For these reasons, it is important to explore and exploit membrane technology to purify crude biodiesel.

  17. Precursor uptake assays and metabolic analyses in isolated tomato fruit chromoplasts

    Directory of Open Access Journals (Sweden)

    Angaman Djédoux


    Full Text Available Abstract Background Carotenoids are the most widespread group of pigments found in nature. In addition to their role in the physiology of the plant, carotenoids also have nutritional relevance as their incorporation in the human diet provides health benefits. In non-photosynthetic tissues, carotenoids are synthesized and stored in specialized plastids called chromoplasts. At present very little is known about the origin of the metabolic precursors and cofactors required to sustain the high rate of carotenoid biosynthesis in these plastids. Recent proteomic data have revealed a number of biochemical and metabolic processes potentially operating in fruit chromoplasts. However, considering that chloroplast to chromoplast differentiation is a very rapid process during fruit ripening, there is the possibility that some of the proteins identified in the proteomic analysis could represent remnants no longer having a functional role in chromoplasts. Therefore, experimental validation is necessary to prove whether these predicted processes are actually operative in chromoplasts. Results A method has been established for high-yield purification of tomato fruit chromoplasts suitable for metabolic studies. Radiolabeled precursors were efficiently incorporated and further metabolized in isolated chromoplast. Analysis of labeled lipophilic compounds has revealed that lipid biosynthesis is a very efficient process in chromoplasts, while the relatively low incorporation levels found in carotenoids suggest that lipid production may represent a competing pathway for carotenoid biosynthesis. Malate and pyruvate are efficiently converted into acetyl-CoA, in agreement with the active operation of the malic enzyme and the pyruvate dehydrogenase complex in the chromoplast. Our results have also shown that isolated chromoplasts can actively sustain anabolic processes without the exogenous supply of ATP, thus suggesting that these organelles may generate this energetic

  18. Quantification of Fusarium oxysporum in fumigated soils by a newly developed real-time PCR assay to assess the efficacy of fumigants for Fusarium wilt disease in strawberry plants. (United States)

    Li, Yuan; Mao, Liangang; Yan, Dongdong; Ma, Taotao; Shen, Jin; Guo, Meixia; Wang, Qiuxia; Ouyang, Canbin; Cao, Aocheng


    Two soil fumigants, chloropicrin (CP) and dimethyl disulfide (DMDS), were used to control Fusarium wilt disease (FWD) which caused large economic losses in strawberries. The fumigants were evaluated alone and in combination in a laboratory study and in strawberry greenhouses. Laboratory tests found that combinations of CP and DMDS indicated a positive synergistic activity on Fusarium oxysporum. A newly developed quantitative assay for F. oxysporum involving real-time PCR was used successfully to evaluate F. oxysporum control by the fumigants; it provided similar results to the selective medium but was less time-consuming and less labor intensive. Greenhouse trials revealed that the combination of CP and DMDS successfully suppressed the incidence of FWD and sharply reduced the population density of F. oxysporum, which significantly increased fruit branch number and maintained a good strawberry yield, higher than methyl bromide (MB) treatment. All of the treatments provided significantly better results than the non-treated control. This study confirms that the newly developed real-time PCR quantitative assay for F. oxysporum was suitable for the control efficacy evaluation of soil fumigants and that the novel fumigant combination of CP and DMDS offers a promising effective alternative to MB for the control of F. oxysporum in strawberry greenhouses. © 2013 Society of Chemical Industry.

  19. Nanotechnology for water treatment and purification

    CERN Document Server

    Apblett, Allen


    This book describes the latest progress in the application of nanotechnology for water treatment and purification. Leaders in the field present both the fundamental science and a comprehensive overview of the diverse range of tools and technologies that have been developed in this critical area. Expert chapters present the unique physicochemical and surface properties of nanoparticles and the advantages that these provide for engineering applications that ensure a supply of safe drinking water for our growing population. Application areas include generating fresh water from seawater, preventing contamination of the environment, and creating effective and efficient methods for remediation of polluted waters. The chapter authors are leading world-wide experts in the field with either academic or industrial experience, ensuring that this comprehensive volume presents the state-of-the-art in the integration of nanotechnology with water treatment and purification. Covers both wastewater and drinking water treatmen...

  20. Purification of nanoparticles by hollow fiber diafiltration (United States)

    Veeken, J.


    Hollow Fiber Diafiltration (Hollow Fiber Tangential Flow Filtration) is an efficient and rapid alternative to traditional methods of nanoparticle purification such as ultracentrifugation, stirred cell filtration, dialysis or chromatography. Hollow Fiber Diafiltration can be used to purify a wide range of nanoparticles including liposomes, colloids, magnetic particles and nanotubes. Hollow Fiber Diafiltration is a membrane based method where pore size determines the retention or transmission of solution components. It is a flow process where the sample is gently circulated through a tubular membrane. With controlled replacement of the permeate or (dialysate), pure nanoparticles can be attained. Hollow Fiber Diafiltration can be directly scaled up from R&D volumes to production. By adding more membrane fibers and maintaining the operating parameters, large volumes can be processed in the same time with the same pressure, and flow dynamics as bench-scale volumes. Keywords: hollow fiber, Diafiltration, filtration, purification, tangential flow filtration.