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Sample records for sybr green qpcr

  1. Identifying Haemophilus haemolyticus and Haemophilus influenzae by SYBR Green real-time PCR.

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    Latham, Roger; Zhang, Bowen; Tristram, Stephen

    2015-05-01

    SYBR Green real time PCR assays for protein D (hpd), fuculose kinase (fucK) and [Cu, Zn]-superoxide dismutase (sodC) were designed for use in an algorithm for the identification of Haemophilus influenzae and H. haemolyticus. When tested on 127 H. influenzae and 60 H. haemolyticus all isolates were identified correctly. Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.

  2. Comparison of the sensitivity of determining soyeabean allergens by ELISA method and SYBR green I

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    Jozef Golian

    2013-07-01

    Full Text Available Normal 0 false false false SK JA X-NONE The aim of this study was to compare the suitability of two methods for detecting defatted soybean powder; SYBR Green I Real-time PCR and enzyme-linked immunosorbent assay (ELISA. Analysis of 20 artificially contaminated samples prepared by simple dilution with wheat flour revealed that both techniques were able to detect defatted soybean powder, although there were significant differences between the two methods. Wheat flour contamination with defatted soybean powder was detected in samples 1-5, (0.012 %; 120 mg.kg-1, but not in samples with lower contamination with soybean powder saples 6-20 using SYBR Green I real-time PCR. Samples 1-10 could not be quantified by ELISA as the absorbance values were greater than the detection limit, and while samples 11-20 were measured, only the values of samples 16, 17 and 18 were within the guaranteed quantification range specified by the ELISA kit manufacturer. Defatted soybean powder contamination was detected in samples 19 and 20, but absorbance values were highly similar to those of the negative control sample.

  3. Development and application of a universal Hemoplasma screening assay based on the SYBR green PCR principle.

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    Willi, Barbara; Meli, Marina L; Lüthy, Ruedi; Honegger, Hanspeter; Wengi, Nicole; Hoelzle, Ludwig E; Reusch, Claudia E; Lutz, Hans; Hofmann-Lehmann, Regina

    2009-12-01

    Hemotropic mycoplasmas (hemoplasmas) are the causative agents of infectious anemia in several mammalian species. Their zoonotic potential has recently been substantiated by the identification of a feline hemoplasma isolate in an immunocompromised human patient. Although species-specific diagnostic molecular methods have been developed, their application as screening tools is limited due to the species diversity of hemoplasmas. The goals of this study were to develop a universal hemoplasma screening assay with broad specificity based on the SYBR green PCR principle, to compare the assay with hemoplasma-specific TaqMan PCR, and to analyze potential tick vectors and human blood samples to address the zoonotic potential. The newly developed PCR assay based on the 16S rRNA gene amplified feline, canine, bovine, porcine, camelid, and murine hemoplasmas, as well as Mycoplasma penetrans and Mycoplasma pneumoniae. The lower detection limit for feline and canine hemoplasmas was 1 to 10 copies/PCR. The assay exhibited 98.2% diagnostic sensitivity and 92.1% diagnostic specificity for feline hemoplasmas. All 1,950 Ixodes ticks were PCR negative, suggesting that Ixodes ticks are not relevant vectors for the above-mentioned hemoplasma species in Switzerland. None of the 414 blood samples derived from anemic or immunocompromised human patients revealed a clear positive result. The SYBR green PCR assay described here is a suitable tool to screen for known and so-far-undiscovered hemoplasma species. Positive results should be confirmed by specific TaqMan PCR or sequencing.

  4. Multiplex real-time PCR SYBR Green for detection and typing of group III Clostridium botulinum.

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    Anniballi, Fabrizio; Auricchio, Bruna; Delibato, Elisabetta; Antonacci, Monia; De Medici, Dario; Fenicia, Lucia

    2012-01-27

    Clostridium botulinum type C and type D belonging to the group III organisms, are mainly responsible for animal botulism outbreaks. Clinical signs alone are often insufficient to make a diagnosis of botulism and a laboratory confirmation is required. Laboratory confirmation can be performed by demonstrating the presence of botulinum neurotoxins in serum, gastrointestinal contents, liver, wound of sick or dead animals, or by demonstrating the presence of C. botulinum in gastrointestinal contents, liver, and wound. Demonstration of spores in gastrointestinal contents or tissue of animals with clinical signs indicative of botulism reinforces the clinical diagnosis. With the aim of detecting and typing C. botulinum group III organisms, a multiplex real-time PCR SYBR Green was developed and in-house validated. Selectivity, limit of detection, relative accuracy, relative specificity, relative sensitivity, and repeatability of the method were investigated. The multiplex real-time PCR SYBR green used showed a 100% selectivity, 100% relative accuracy, 100% relative specificity, 100% relative sensitivity and a limit of detection of 277 and 580 DNA copies for C. botulinum type C and C. botulinum type D, respectively. The method reported here represents a suitable tool for laboratory diagnosis of type C and D botulism and for testing a large number of samples collected during the animal botulism surveillance and prevention activities. Copyright © 2011 Elsevier B.V. All rights reserved.

  5. Application of silver nanoparticles in the detection of SYBR Green I by surface enhanced Raman and surface-enhanced fluorescence

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    Guo, Wei; Wu, Jian; Wang, Chunyan; Zhang, Tian; Chen, Tao

    2018-05-01

    Silver nanomaterials have remarkable application in biomedical detection due to their unique surface plasmon resonance (SPR) characteristics. It can be used for surface-enhanced Raman scattering (SERS) and surface-enhanced fluorescence (SEF). Current research elaborates a technique for improvement of SYBR Green I detection obtained from surface-enhanced Raman scattering (SERS) and surface-enhanced fluorescence (SEF) by silver nanoparticles with the average size about 70 nm. Primarily, SYBR Green I is an important fluorescent dye used in polymerase chain reaction (PCR). It is found that both Raman and fluorescence can be used for detection of this dye. Furthermore, the enhanced efficiency of the Raman and fluorescence by SERS and SEF is observed in this study, the enhancement factor for Raman signals is 3.2 × 103, and the fluorescence intensity bincreased two times by SEF. The quantitative detection of SYBR Green I by SERS and SEF can be achieved. The present work can be used to improve the detection of SYBR Green I by SERS and SEF. It would also be employed for high-sensitive detection of other materials in the future.

  6. SYBR green-based detection of Leishmania infantum DNA using peripheral blood samples.

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    Ghasemian, Mehrdad; Gharavi, Mohammad Javad; Akhlaghi, Lame; Mohebali, Mehdi; Meamar, Ahmad Reza; Aryan, Ehsan; Oormazdi, Hormozd; Ghayour, Zahra

    2016-03-01

    Parasitological methods for the diagnosis of visceral leishmaniasis (VL) require invasive sampling procedures. The aim of this study was to detect Leishmania infantum (L. infantum) DNA by real time-PCR method in peripheral blood of symptomatic VL patient and compared its performance with nested PCR, an established molecular method with very high diagnostic indices. 47 parasitologically confirmed VL patients diagnosed by direct agglutination test (DAT > 3200), bone marrow aspiration and presented characteristic clinical features (fever, hepatosplenomegaly, and anemia) and 40 controls (non-endemic healthy control-30, Malaria-2, Toxoplasma gondii-2, Mycobacterium tuberculosis-2, HBV-1, HCV-1, HSV-1 and CMV-1) were enrolled in this study. SYBR-green based real time-PCR and nested PCR was performed to amplify the Kinetoplast DNA minicircle gene using the DNA extracted from Buffy coat. From among 47 patients, 45 (95.7 %) were positive by both nested-PCR and real time-PCR. These results indicate that real time-PCR was not only as sensitive as a nested-PCR assay for detection of Leishmania kDNA in clinical sample, but also more rapid. The advantage of real time-PCR based methods over nested-PCR is simple to perform, more faster in which nested-PCR requires post-PCR processing and reducing contamination risk.

  7. Detection and characterization of Leishmania (Leishmania and Leishmania (Viannia by SYBR green-based real-time PCR and high resolution melt analysis targeting kinetoplast minicircle DNA.

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    Marcello Ceccarelli

    Full Text Available Leishmaniasis is a neglected disease with a broad clinical spectrum which includes asymptomatic infection. A thorough diagnosis, able to distinguish and quantify Leishmania parasites in a clinical sample, constitutes a key step in choosing an appropriate therapy, making an accurate prognosis and performing epidemiological studies. Several molecular techniques have been shown to be effective in the diagnosis of leishmaniasis. In particular, a number of PCR methods have been developed on various target DNA sequences including kinetoplast minicircle constant regions. The first aim of this study was to develop a SYBR green-based qPCR assay for Leishmania (Leishmania infantum detection and quantification, using kinetoplast minicircle constant region as target. To this end, two assays were compared: the first used previously published primer pairs (qPCR1, whereas the second used a nested primer pairs generating a shorter PCR product (qPCR2. The second aim of this study was to evaluate the possibility to discriminate among subgenera Leishmania (Leishmania and Leishmania (Viannia using the qPCR2 assay followed by melting or High Resolution Melt (HRM analysis. Both assays used in this study showed good sensitivity and specificity, and a good correlation with standard IFAT methods in 62 canine clinical samples. However, the qPCR2 assay allowed to discriminate between Leishmania (Leishmania and Leishmania (Viannia subgenera through melting or HRM analysis. In addition to developing assays, we investigated the number and genetic variability of kinetoplast minicircles in the Leishmania (L. infantum WHO international reference strain (MHOM/TN/80/IPT1, highlighting the presence of minicircle subclasses and sequence heterogeneity. Specifically, the kinetoplast minicircle number per cell was estimated to be 26,566±1,192, while the subclass of minicircles amplifiable by qPCR2 was estimated to be 1,263±115. This heterogeneity, also observed in canine clinical

  8. An Evaluation of Quantitative PCR Assays (TaqMan® and SYBR Green for the Detection of Babesia bigemina and Babesia bovis, and a Novel Fluorescent-ITS1-PCR Capillary Electrophoresis Method for Genotyping B. bovis Isolates

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    Bing Zhang

    2016-09-01

    Full Text Available Babesia spp. are tick-transmitted haemoparasites causing tick fever in cattle. In Australia, economic losses to the cattle industry from tick fever are estimated at AUD$26 Million per annum. If animals recover from these infections, they become immune carriers. Here we describe a novel multiplex TaqMan qPCR targeting cytochrome b genes for the identification of Babesia spp. The assay shows high sensitivity, specificity and reproducibility, and allows quantification of parasite DNA from Babesia bovis and B. bigemina compared to standard PCR assays. A previously published cytochrome b SYBR Green qPCR was also tested in this study, showing slightly higher sensitivity than the Taqman qPCRs but requires melting curve analysis post-PCR to confirm specificity. The SYBR Green assays were further evaluated using both diagnostic submissions and vaccinated cattle (at 7, 9, 11 and 14 days post-inoculation showed that B. bigemina can be detected more frequently than B. bovis. Due to fewer circulating parasites, B. bovis detection in carrier animals requires higher DNA input. Preliminary data for a novel fluorescent PCR genotyping based on the Internal Transcribed Spacer 1 region to detect vaccine and field alleles of B. bovis are described. This assay is capable of detecting vaccine and novel field isolate alleles in a single sample.

  9. Detection and Differentiation of Leishmania spp. in Clinical Specimens by Use of a SYBR Green-Based Real-Time PCR Assay.

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    de Almeida, Marcos E; Koru, Ozgur; Steurer, Francis; Herwaldt, Barbara L; da Silva, Alexandre J

    2017-01-01

    Leishmaniasis in humans is caused by Leishmania spp. in the subgenera Leishmania and Viannia Species identification often has clinical relevance. Until recently, our laboratory relied on conventional PCR amplification of the internal transcribed spacer 2 (ITS2) region (ITS2-PCR) followed by sequencing analysis of the PCR product to differentiate Leishmania spp. Here we describe a novel real-time quantitative PCR (qPCR) approach based on the SYBR green technology (LSG-qPCR), which uses genus-specific primers that target the ITS1 region and amplify DNA from at least 10 Leishmania spp., followed by analysis of the melting temperature (T m ) of the amplicons on qPCR platforms (the Mx3000P qPCR system [Stratagene-Agilent] and the 7500 real-time PCR system [ABI Life Technologies]). We initially evaluated the assay by testing reference Leishmania isolates and comparing the results with those from the conventional ITS2-PCR approach. Then we compared the results from the real-time and conventional molecular approaches for clinical specimens from 1,051 patients submitted to the reference laboratory of the Centers for Disease Control and Prevention for Leishmania diagnostic testing. Specimens from 477 patients tested positive for Leishmania spp. with the LSG-qPCR assay, specimens from 465 of these 477 patients also tested positive with the conventional ITS2-PCR approach, and specimens from 10 of these 465 patients had positive results because of retesting prompted by LSG-qPCR positivity. On the basis of the T m values of the LSG-qPCR amplicons from reference and clinical specimens, we were able to differentiate four groups of Leishmania parasites: the Viannia subgenus in aggregate; the Leishmania (Leishmania) donovani complex in aggregate; the species L (L) tropica; and the species L (L) mexicana, L (L) amazonensis, L (L) major, and L (L) aethiopica in aggregate. Copyright © 2016 American Society for Microbiology.

  10. Effect of acidic pH on flow cytometric detection of bacteria stained with SYBR Green I and their distinction from background

    International Nuclear Information System (INIS)

    Baldock, Daniel; Nocker, Andreas; Nebe-von-Caron, Gerhard; Bongaerts, Roy

    2013-01-01

    Unspecific background caused by biotic or abiotic particles, cellular debris, or autofluorescence is a well-known interfering parameter when applying flow cytometry to the detection of microorganisms in combination with fluorescent dyes. We present here an attempt to suppress the background signal intensity and thus to improve the detection of microorganisms using the nucleic acid stain SYBR ® Green I. It has been observed that the fluorescent signals from SYBR Green I are greatly reduced at acidic pH. When lowering the pH of pre-stained samples directly prior to flow cytometric analysis, we hypothesized that the signals from particles and cells with membrane damage might therefore be reduced. Signals from intact cells, temporarily maintaining a neutral cytosolic pH, should not be affected. We show here that this principle holds true for lowering background interference, whereas the signals of membrane-compromised dead cells are only affected weakly. Signals from intact live cells at low pH were mostly comparable to signals without acidification. Although this study was solely performed with SYBR ® Green I, the principle of low pH flow cytometry (low pH-FCM) might hold promise when analyzing complex matrices with an abundance of non-cellular matter, especially when expanded to non-DNA binding dyes with a stronger pH dependence of fluorescence than SYBR Green I and a higher pK a value. (paper)

  11. Comparison of nested-multiplex, Taqman & SYBR Green real-time PCR in diagnosis of amoebic liver abscess in a tertiary health care institute in India.

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    Dinoop, K P; Parija, Subhash Chandra; Mandal, Jharna; Swaminathan, R P; Narayanan, P

    2016-01-01

    Amoebiasis is a common parasitic infection caused by Entamoeba histolytica and amoebic liver abscess (ALA) is the most common extraintestinal manifestation of amoebiasis. The aim of this study was to standardise real-time PCR assays (Taqman and SYBR Green) to detect E. histolytica from liver abscess pus and stool samples and compare its results with nested-multiplex PCR. Liver abscess pus specimens were subjected to DNA extraction. The extracted DNA samples were subjected to amplification by nested-multiplex PCR, Taqman (18S rRNA) and SYBR Green real-time PCR (16S-like rRNA assays to detect E. histolytica/E. dispar/E. moshkovskii). The amplification products were further confirmed by DNA sequence analysis. Receiver operator characteristic (ROC) curve analysis was done for nested-multiplex and SYBR Green real-time PCR and the area under the curve was calculated for evaluating the accuracy of the tests to dignose ALA. In all, 17, 19 and 25 liver abscess samples were positive for E. histolytica by nested-multiplex PCR, SYBR Green and Taqman real-time PCR assays, respectively. Significant differences in detection of E. histolytica were noted in the real-time PCR assays evaluated ( Pnested-multiplex PCR, SYBR Green real-time PCR and Taqman real-time PCR evaluated showed a positivity rate of 34, 38 and 50 per cent, respectively. Based on ROC curve analysis (considering Taqman real-time PCR as the gold standard), it was observed that SYBR Green real-time PCR was better than conventional nested-multiplex PCR for the diagnosis of ALA. Taqman real-time PCR targeting the 18S rRNA had the highest positivity rate evaluated in this study. Both nested multiplex and SYBR Green real-time PCR assays utilized were evaluated to give accurate results. Real-time PCR assays can be used as the gold standard in rapid and reliable diagnosis, and appropriate management of amoebiasis, replacing the conventional molecular methods.

  12. Quantification of ochratoxin A-producing molds in food products by SYBR Green and TaqMan real-time PCR methods

    DEFF Research Database (Denmark)

    Rodríguez, Alicia; Rodríguez, Mar; Luque, M. Isabel

    2011-01-01

    , usually reported in food products, were used as references. All strains were tested for OTA production by mycellar electrokinetic capillary electrophoresis (MECE) and high-pressure liquid chromatography-mass spectrometry (HPLC-MS). The ability of the optimized qPCR protocols to quantify OTA......Ochratoxin A (OTA) is a mycotoxin synthesized by a variety of different fungi, most of them from the genera Penicillium and Aspergillus. Early detection and quantification of OTA producing species is crucial to improve food safety. In the present work, two protocols of real-time qPCR based on SYBR......-producing molds was evaluated in different artificially inoculated foods. A good linear correlation was obtained over the range 1 x 104 to 10 conidia/g per reaction for all qPCR assays in the different food matrices (cooked and cured products and fruits). The detection limit in all inoculated foods ranged between...

  13. Cross-platform evaluation of commercial real-time SYBR green RT-PCR kits for sensitive and rapid detection of European bat Lyssavirus type 1.

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    Picard-Meyer, Evelyne; Peytavin de Garam, Carine; Schereffer, Jean Luc; Marchal, Clotilde; Robardet, Emmanuelle; Cliquet, Florence

    2015-01-01

    This study evaluates the performance of five two-step SYBR Green RT-qPCR kits and five one-step SYBR Green qRT-PCR kits using real-time PCR assays. Two real-time thermocyclers showing different throughput capacities were used. The analysed performance evaluation criteria included the generation of standard curve, reaction efficiency, analytical sensitivity, intra- and interassay repeatability as well as the costs and the practicability of kits, and thermocycling times. We found that the optimised one-step PCR assays had a higher detection sensitivity than the optimised two-step assays regardless of the machine used, while no difference was detected in reaction efficiency, R (2) values, and intra- and interreproducibility between the two methods. The limit of detection at the 95% confidence level varied between 15 to 981 copies/µL and 41 to 171 for one-step kits and two-step kits, respectively. Of the ten kits tested, the most efficient kit was the Quantitect SYBR Green qRT-PCR with a limit of detection at 95% of confidence of 20 and 22 copies/µL on the thermocyclers Rotor gene Q MDx and MX3005P, respectively. The study demonstrated the pivotal influence of the thermocycler on PCR performance for the detection of rabies RNA, as well as that of the master mixes.

  14. Cross-Platform Evaluation of Commercial Real-Time SYBR Green RT-PCR Kits for Sensitive and Rapid Detection of European Bat Lyssavirus Type 1

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    Evelyne Picard-Meyer

    2015-01-01

    Full Text Available This study evaluates the performance of five two-step SYBR Green RT-qPCR kits and five one-step SYBR Green qRT-PCR kits using real-time PCR assays. Two real-time thermocyclers showing different throughput capacities were used. The analysed performance evaluation criteria included the generation of standard curve, reaction efficiency, analytical sensitivity, intra- and interassay repeatability as well as the costs and the practicability of kits, and thermocycling times. We found that the optimised one-step PCR assays had a higher detection sensitivity than the optimised two-step assays regardless of the machine used, while no difference was detected in reaction efficiency, R2 values, and intra- and interreproducibility between the two methods. The limit of detection at the 95% confidence level varied between 15 to 981 copies/µL and 41 to 171 for one-step kits and two-step kits, respectively. Of the ten kits tested, the most efficient kit was the Quantitect SYBR Green qRT-PCR with a limit of detection at 95% of confidence of 20 and 22 copies/µL on the thermocyclers Rotor gene Q MDx and MX3005P, respectively. The study demonstrated the pivotal influence of the thermocycler on PCR performance for the detection of rabies RNA, as well as that of the master mixes.

  15. Development of melting temperature-based SYBR Green I polymerase chain reaction methods for multiplex genetically modified organism detection.

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    Hernández, Marta; Rodríguez-Lázaro, David; Esteve, Teresa; Prat, Salomé; Pla, Maria

    2003-12-15

    Commercialization of several genetically modified crops has been approved worldwide to date. Uniplex polymerase chain reaction (PCR)-based methods to identify these different insertion events have been developed, but their use in the analysis of all commercially available genetically modified organisms (GMOs) is becoming progressively insufficient. These methods require a large number of assays to detect all possible GMOs present in the sample and thereby the development of multiplex PCR systems using combined probes and primers targeted to sequences specific to various GMOs is needed for detection of this increasing number of GMOs. Here we report on the development of a multiplex real-time PCR suitable for multiple GMO identification, based on the intercalating dye SYBR Green I and the analysis of the melting curves of the amplified products. Using this method, different amplification products specific for Maximizer 176, Bt11, MON810, and GA21 maize and for GTS 40-3-2 soybean were obtained and identified by their specific Tm. We have combined amplification of these products in a number of multiplex reactions and show the suitability of the methods for identification of GMOs with a sensitivity of 0.1% in duplex reactions. The described methods offer an economic and simple alternative to real-time PCR systems based on sequence-specific probes (i.e., TaqMan chemistry). These methods can be used as selection tests and further optimized for uniplex GMO quantification.

  16. Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR

    DEFF Research Database (Denmark)

    Hillig, Ann-Britt Nygaard; Jørgensen, Claus Bøttcher; Cirera, Susanna

    2007-01-01

    -microglobulin (B2M), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hydroxymethylbilane synthase (HMBS), hypoxanthine phosphoribosyltransferase I (HPRT I), ribosomal protein L4 (RPL4), succinate dehydrogenase complex subunit A (SDHA), TATA box binding protein (TPB) and tyrosine 3-monooxygenase/tryptophan 5......-monooxygenase activation protein zeta polypeptide (YWHAZ). The stability of these reference genes in different pig tissues was investigated using the geNorm application. The range of expression stability in the genes analysed was (from the most stable to the least stable): ACTB/RPL4, TBP, HPRT, HMBS, YWHAZ...

  17. Optimization of the Sybr Green real time PCR for the detection of Human Herpes Virus type 6 (HHV-6 Otimização da PCR em tempo real - Sybr Green para detecção do Herpes Vírus Humano tipo 6 (HHV-6

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    Cynthia Liliane Motta do Canto

    2008-02-01

    Full Text Available HHV-6 is the etiological agent of Exanthem subitum which is considered the sixth most frequent disease in infancy. In immuno-compromised hosts, reactivation of latent HHV-6 infection may cause severe acute disease. We developed a Sybr Green Real Time PCR for HHV-6 and compared the results with nested conventional PCR. A 214 pb PCR derived fragment was cloned using pGEM-T easy from Promega system. Subsequently, serial dilutions were made in a pool of negative leucocytes from 10-6 ng/µL (equivalent to 2465.8 molecules/µL to 10-9 (equivalent to 2.46 molecules/µL. Dilutions of the plasmid were amplified by Sybr Green Real Time PCR, using primers HHV3 (5' TTG TGC GGG TCC GTT CCC ATC ATA 3'and HHV4 (5' TCG GGA TAG AAA AAC CTA ATC CCT 3' and by conventional nested PCR using primers HHV1 (outer: 5'CAA TGC TTT TCT AGC CGC CTC TTC 3'; HHV2 (outer: 5' ACA TCT ATA ATT TTA GAC GAT CCC 3'; HHV3 (inner and HHV4 (inner 3'. The detection threshold was determined by plasmid serial dilutions. Threshold for Sybr Green real time PCR was 24.6 molecules/µL and for the nested PCR was 2.46 molecules/µL. We chose the Real Time PCR for diagnosing and quantifying HHV-6 DNA from samples using the new Sybr Green chemistry due to its sensitivity and lower risk of contamination.HHV-6 é o agente etiológico do Exantema Súbito e considerado a sexta doença mais comum na infância. Em indivíduos imunocomprometidos, a reativação da infecção latente pode causar doença aguda ou morte. Padronizamos PCR em Tempo Real utilizando a química Sybr Green na detecção do HHV-6 e comparamos os resultados com a PCR convencional. Um fragmento de 214 pb foi clonado através do kit pGEM-T do sistema Promega. Com este clone, foram feitas diluições seriadas em um pool de leucócitos negativos a partir de 10-6 ng/µL (equivalente a 2465,8 moleculas/µL até 10-9 (equivalente a 2,46 moleculas/µL. As diluições foram amplificadas por PCR em Tempo Real utilizando Sybr Green, com

  18. Development of SYBR Green and TaqMan quantitative real-time PCR assays for hepatopancreatic parvovirus (HPV) infecting Penaeus monodon in India.

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    Yadav, Reena; Paria, Anutosh; Mankame, Smruti; Makesh, M; Chaudhari, Aparna; Rajendran, K V

    2015-12-01

    Hepatopancreatic parvovirus (HPV) infects Penaeus monodon and causes mortality in the larval stages. Further, it has been implicated in the growth retardation in cultured P. monodon. Though different geographical isolates of HPV show large sequence variations, a sensitive PCR assay specific to Indian isolate has not yet been reported. Here, we developed a sensitive SYBR Green-based and TaqMan real-time PCR for the detection and quantification of the virus. A 441-bp PCR amplicon was cloned in pTZ57 R/T vector and the plasmid copy number was estimated. A 10-fold serial dilution of the plasmid DNA from 1 × 10(9) copies to 1 copy was prepared and used as the standard. The primers were tested initially using the standard on a conventional PCR format to determine the linearity of detection. The standards were further tested on real-time PCR format using SYBR Green and TaqMan chemistry and standard curves were generated based on the Ct values from three well replicates for each dilution. The assays were found to be sensitive, specific and reproducible with a wide dynamic range (1 × 10(9) to 10 copies) with coefficient of regression (R(2)) > 0.99, calculated average slope -3.196 for SYBR Green assay whereas, for TaqMan assay it was >0.99 and -3.367, respectively. The intra- and inter-assay variance of the Ct values ranged from 0.26% to 0.94% and 0.12% to 0.81%, respectively, for SYBR Green assay, and the inter-assay variance of the Ct values for TaqMan assay ranged from 0.07% to 1.93%. The specificity of the assays was proved by testing other DNA viruses of shrimp such as WSSV, IHHNV and MBV. Standardized assays were further tested to detect and quantify HPV in the post-larvae of P. monodon. The result was further compared with conventional PCR to test the reproducibility of the test. The assay was also used to screen Litopeneaus vannamei, Macrobrachium rosenbergii and Scylla serrata for HPV. Copyright © 2015 Elsevier Ltd. All rights reserved.

  19. Development and validation of a SYBR Green I-based real-time polymerase chain reaction method for detection of haptoglobin gene deletion in clinical materials.

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    Soejima, Mikiko; Tsuchiya, Yuji; Egashira, Kouichi; Kawano, Hiroyuki; Sagawa, Kimitaka; Koda, Yoshiro

    2010-06-01

    Anhaptoglobinemic patients run the risk of severe anaphylactic transfusion reaction because they produce serum haptoglobin (Hp) antibodies. Being homozygous for the Hp gene deletion (HP(del)) is the only known cause of congenital anhaptoglobinemia, and clinical diagnosis of HP(del) before transfusion is important to prevent anaphylactic shock. We recently developed a 5'-nuclease (TaqMan) real-time polymerase chain reaction (PCR) method. A SYBR Green I-based duplex real-time PCR assay using two forward primers and a common reverse primer followed by melting curve analysis was developed to determine HP(del) zygosity in a single tube. In addition, to obviate initial DNA extraction, we examined serially diluted blood samples as PCR templates. Allelic discrimination of HP(del) yielded optimal results at blood sample dilutions of 1:64 to 1:1024. The results from 2231 blood samples were fully concordant with those obtained by the TaqMan-based real-time PCR method. The detection rate of the HP(del) allele by the SYBR Green I-based method is comparable with that using the TaqMan-based method. This method is readily applicable due to its low initial cost and analyzability using economical real-time PCR machines and is suitable for high-throughput analysis as an alternative method for allelic discrimination of HP(del).

  20. Quantitative detection of the potato cyst nematode, Globodera pallida, and the beet cyst nematode, Heterodera schachtii, using Real-Time PCR with SYBR green I dye.

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    Madani, Mehrdad; Subbotin, Sergei A; Moens, Maurice

    2005-04-01

    The potato cyst nematode Globodera pallida and the beet cyst nematode Heterodera schachtii are major nematode pests in world agriculture. Precise identification and knowledge about the number of nematodes in field soil are necessary to develop effective integrated pest control. Here we report the results of the Real-Time PCR assay for the rapid detection and quantification of G. pallida and H. schachtii. Using species specific primers and SYBR green I dye, we were able to detect a single second stage juvenile of cyst forming nematodes in samples. The specificity of the reaction was confirmed by the lack of amplification of DNAs from other Heterodera or Globodera species. Validation tests showed a rather high correlation between real numbers of second stage juveniles in a sample and expected numbers detected by Real-Time PCR. Reasons for observed differences in sensitivity and reliability of quantification detection for two species as well as other problems of Real-Time PCR are discussed. The Real-Time PCR assay with SYBR green I dye targeting fragments of the ITS-rDNA provided a sensitive means for the rapid and simultaneous detection and quantification of juveniles of these pests.

  1. An Undergraduate Laboratory Experiment for Upper-Level Forensic Science, Biochemistry, or Molecular Biology Courses: Human DNA Amplification Using STR Single Locus Primers by Real-Time PCR with SYBR Green Detection

    Science.gov (United States)

    Elkins, Kelly M.; Kadunc, Raelynn E.

    2012-01-01

    In this laboratory experiment, real-time polymerase chain reaction (real-time PCR) was conducted using published human TPOX single-locus DNA primers for validation and various student-designed short tandem repeat (STR) primers for Combined DNA Index System (CODIS) loci. SYBR Green was used to detect the amplification of the expected amplicons. The…

  2. Detection and discrimination of five E. coli pathotypes using a combinatory SYBR® Green qPCR screening system.

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    Barbau-Piednoir, Elodie; Denayer, Sarah; Botteldoorn, Nadine; Dierick, Katelijne; De Keersmaecker, Sigrid C J; Roosens, Nancy H

    2018-04-01

    A detection and discrimination system for five Escherichia coli pathotypes, based on a combination of 13 SYBR® Green qPCR, has been developed, i.e., combinatory SYBR® Green qPCR screening system for pathogenic E. coli (CoSYPS Path E. coli). It allows the discrimination on isolates and the screening of potential presence in food of the following pathotypes of E. coli: shigatoxigenic (STEC) (including enterohemorrhagic (EHEC)), enteropathogenic (EPEC), enteroaggregative (EAggEC), enteroaggregative shigatoxigenic (EAggSTEC), and enteroinvasive (EIEC) E. coli. The SYBR® Green qPCR assays target the uidA, ipaH, eae, aggR, aaiC, stx1, and stx2 genes. uidA controls for E. coli presence and all the other genes are specific targets of E. coli pathotypes. For each gene, two primer pairs have been designed to guarantee a sufficient detection even in case of deletion or polymorphisms in the target gene. Moreover, all the qPCR have been designed to be run together in a single analytical PCR plate. This study includes the primer pairs' design, in silico and in situ selectivity, sensitivity, repeatability, and reproducibility evaluation of the 13 SYBR® Green qPCR assays. Each target displayed a selectivity of 100%. The limit of detection of the 13 assays is between 1 and 10 genomic copies. Their repeatability and reproducibility comply with the European requirements. As a preliminary feasibility study on food, the CoSYPS Path E. coli system was subsequently evaluated on four food matrices artificially contaminated with pathogenic E. coli. It allowed the detection of an initial contamination level as low as 2 to 7 cfu of STEC/25 g of food matrix after 24 h of enrichment.

  3. Real-time PCR Detection of Brucella Abortus: A Comparative Study of SYBR Green I, 5'-exonuclease, and Hybridization Probe Assays

    Energy Technology Data Exchange (ETDEWEB)

    Newby, Deborah Trishelle; Hadfield, Ted; Roberto, Francisco Figueroa

    2003-08-01

    Real-time PCR provides a means of detecting and quantifying DNA targets by monitoring PCR product accumulation during cycling as indicated by increased fluorescence. A number of different approaches can be used to generate the fluorescence signal. Three approaches—SYBR Green I (a double-stranded DNA intercalating dye), 5'-exonuclease (enzymatically released fluors), and hybridization probes (fluorescence resonance energy transfer)—were evaluated for use in a real-time PCR assay to detect Brucella abortus. The three assays utilized the same amplification primers to produce an identical amplicon. This amplicon spans a region of the B. abortus genome that includes portions of the alkB gene and the IS711 insertion element. All three assays were of comparable sensitivity, providing a linear assay over 7 orders of magnitude (from 7.5 ng down to 7.5 fg). However, the greatest specificity was achieved with the hybridization probe assay.

  4. A Multiplex SYBR Green Real-Time PCR Assay for the Detection of Three Colistin Resistance Genes from Cultured Bacteria, Feces, and Environment Samples

    Directory of Open Access Journals (Sweden)

    Jiyun Li

    2017-10-01

    Full Text Available The aim of the study was to develop a multiplex assay for rapid detection of mcr-1, mcr-2, and mcr-3, a group of genes of conferring resistance to colistin mediated by plasmid in Enterobacteriaceae. A SYBR Green based real-time PCR assay has been designed to detect the mcr genes, and applied to cultured bacteria, feces and soil samples. All three mcr genes could be detected with a lower limit of 102 cultured bacteria. This test was highly specific and sensitive, and generated no false-positive results. The assay was also conclusive when applied to feces and soil samples containing mcr-1-positive Escherichia coli, which could facilitate the screening of mcr genes not only in the bacteria, but also directly from the environment. This simple, rapid, sensitive, and specific multiplex assay will be useful for rapid screening of the colistin resistance in both clinical medicine and animal husbandry.

  5. Modulation of the Singlet Oxygen Generation from the Double Strand DNA-SYBR Green I Complex Mediated by T-Melamine-T Mismatch for Visual Detection of Melamine.

    Science.gov (United States)

    Hu, Hao; Zhang, Jinyi; Ding, Yu; Zhang, Xinfeng; Xu, Kailai; Hou, Xiandeng; Wu, Peng

    2017-05-02

    Singlet oxygen ( 1 O 2 ), generated via photosensitization, has been proved to oxidize chromogenic substrates with neither H 2 O 2 oxidation nor enzyme (horseradish peroxidase, HRP) catalysis. Of the various methods for modulation of the 1 O 2 generation, DNA-controlled photosensitization received great attention. Therefore, integration of the formation/deformation DNA structures with DNA-controlled photosensitization will be extremely appealing in visual biosensor developments. Here, the stable melamine-thymine complex was explored in combination with DNA-controlled photosensitization for visual detection of melamine. A T-rich single stand DNA was utilized as the recognition unit. Upon the formation of the T-M-T complex, double stand DNA was formed, which was ready for the binding of SYBR Green I and activated the photosensitization. Subsequent oxidation of TMB allowed visual detection of melamine in dairy products, with spike-recoveries ranging from 94% to 106%.

  6. Rapid and Specific Detection of Acidovorax avenae subsp. citrulli Using SYBR Green-Based Real-Time PCR Amplification of the YD-Repeat Protein Gene.

    Science.gov (United States)

    Cho, Min Seok; Park, Duck Hwan; Ahn, Tae-Young; Park, Dong Suk

    2015-09-01

    The aim of this study was to develop a SYBR Green-based real-time PCR assay for the rapid, specific, and sensitive detection of Acidovorax avenae subsp. citrulli, which causes bacterial fruit blotch (BFB), a serious disease of cucurbit plants. The molecular and serological methods currently available for the detection of this pathogen are insufficiently sensitive and specific. Thus, a novel SYBR Green-based real-time PCR assay targeting the YD-repeat protein gene of A. avenae subsp. citrulli was developed. The specificity of the primer set was evaluated using DNA purified from 6 isolates of A. avenae subsp. citrulli, 7 other Acidovorax species, and 22 of non-targeted strains, including pathogens and non-pathogens. The AC158F/R primer set amplified a single band of the expected size from genomic DNA obtained from the A. avenae subsp. citrulli strains but not from the genomic DNA of other Acidovorax species, including that of other bacterial genera. Using this assay, it was possible to detect at least one genomeequivalents of the cloned amplified target DNA using 5 × 10(0) fg/μl of purified genomic DNA per reaction or using a calibrated cell suspension, with 6.5 colony-forming units per reaction being employed. In addition, this assay is a highly sensitive and reliable method for identifying and quantifying the target pathogen in infected samples that does not require DNA extraction. Therefore, we suggest that this approach is suitable for the rapid and efficient diagnosis of A. avenae subsp. citrulli contaminations of seed lots and plants.

  7. Comparison of real-time SYBR green dengue assay with real-time taqman RT-PCR dengue assay and the conventional nested PCR for diagnosis of primary and secondary dengue infection

    Science.gov (United States)

    Paudel, Damodar; Jarman, Richard; Limkittikul, Kriengsak; Klungthong, Chonticha; Chamnanchanunt, Supat; Nisalak, Ananda; Gibbons, Robert; Chokejindachai, Watcharee

    2011-01-01

    Background: Dengue fever and dengue hemorrhagic fever are caused by dengue virus. Dengue infection remains a burning problem of many countries. To diagnose acute dengue in the early phase we improve the low cost, rapid SYBR green real time assay and compared the sensitivity and specificity with real time Taqman® assay and conventional nested PCR assay. Aims: To develop low cost, rapid and reliable real time SYBR green diagnostic dengue assay and compare with Taqman real-time assay and conventional nested PCR (modified Lanciotti). Materials and Methods: Eight cultured virus strains were diluted in tenth dilution down to undetectable level by the PCR to optimize the primer, temperature (annealing, and extension and to detect the limit of detection of the assay. Hundred and ninety three ELISA and PCR proved dengue clinical samples were tested with real time SYBR® Green assay, real time Taqman® assay to compare the sensitivity and specificity. Results: Sensitivity and specificity of real time SYBR® green dengue assay (84% and 66%, respectively) was almost comparable to those (81% and 74%) of Taqman real time PCR dengue assay. Real time SYBR® green RT-PCR was equally sensitive in primary and secondary infection while real time Taqman was less sensitive in the secondary infection. Sensitivity of real time Taqman on DENV3 (87%) was equal to SYBR green real time PCR dengue assay. Conclusion: We developed low cost rapid diagnostic SYBR green dengue assay. Further study is needed to make duplex primer assay for the serotyping of dengue virus. PMID:22363089

  8. Development of loop-mediated isothermal amplification and SYBR green real-time PCR methods for the detection of Citrus yellow mosaic badnavirus in citrus species.

    Science.gov (United States)

    Anthony Johnson, A M; Dasgupta, I; Sai Gopal, D V R

    2014-07-01

    Citrus yellow mosaic badnavirus (CMBV) is an important pathogen in southern India spread by infected citrus propagules. One of the measures to arrest the spread of CMBV is to develop methods to screen and certify citrus propagules as CMBV-free. The methods loop-mediated isothermal amplification (LAMP) and SYBR green real-time PCR (SGRTPCR) have been developed for the efficient detection of CMBV in citrus propagules. This paper compares the sensitivities of LAMP and SGRTPCR with polymerase chain reaction (PCR) for the detection of CMBV. Whereas PCR and LAMP were able to detect CMBV from a minimum of 10 ng of total DNA of infected leaf samples, SGRTPCR could detect the same from 1 ng of total DNA. Using SGRTPCR, the viral titres were estimated to be the highest in rough lemon and lowest in Nagpur Mandarin of the five naturally infected citrus species tested. The results will help in designing suitable strategies for the sensitive detection of CMBV from citrus propagules. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. SYBR green-based one step quantitative real-time polymerase chain reaction assay for the detection of Zika virus in field-caught mosquitoes.

    Science.gov (United States)

    Tien, Wei-Ping; Lim, Gareth; Yeo, Gladys; Chiang, Suzanna Nicole; Chong, Chee-Seng; Ng, Lee-Ching; Hapuarachchi, Hapuarachchige Chanditha

    2017-09-19

    The monitoring of vectors is one of the key surveillance measures to assess the risk of arbovirus transmission and the success of control strategies in endemic regions. The recent re-emergence of Zika virus (ZIKV) in the tropics, including Singapore, emphasizes the need to develop cost-effective, rapid and accurate assays to monitor the virus spread by mosquitoes. As ZIKV infections largely remain asymptomatic, early detection of ZIKV in the field-caught mosquitoes enables timely implementation of appropriate mosquito control measures. We developed a rapid, sensitive and specific real-time reverse transcription polymerase chain reaction (rRT-PCR) assay for the detection of ZIKV in field-caught mosquitoes. The primers and PCR cycling conditions were optimized to minimize non-specific amplification due to cross-reactivity with the genomic material of Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, Culex tritaeniorhynchus, Culex sitiens and Anopheles sinensis, as well as accompanying microbiota. The performance of the assay was further evaluated with a panel of flaviviruses and alphaviruses as well as in field-caught Ae. aegypti mosquitoes confirmed to be positive for ZIKV. As compared to a probe-based assay, the newly developed assay demonstrated 100% specificity and comparable detection sensitivity for ZIKV in mosquitoes. Being a SYBR Green-based method, the newly-developed assay is cost-effective and easy to adapt, thus is applicable to large-scale vector surveillance activities in endemic countries, including those with limited resources and expertise. The amplicon size (119 bp) also allows sequencing to confirm the virus type. The primers flank relatively conserved regions of ZIKV genome, so that, the assay is able to detect genetically diverse ZIKV strains. Our findings, therefore, testify the potential use of the newly-developed assay in vector surveillance programmes for ZIKV in endemic regions.

  10. Evaluation and optimization of SYBR Green real-time reverse transcription polymerase chain reaction as a tool for diagnosis of the Flavivirus genus in Brazil

    Directory of Open Access Journals (Sweden)

    Marilia Farignoli Romeiro

    2016-06-01

    Full Text Available Abstract: INTRODUCTION: The genus Flavivirus includes several pathogenic species that cause severe illness in humans. Therefore, a rapid and accurate molecular method for diagnosis and surveillance of these viruses would be of great importance. Here, we evaluate and optimize a quantitative real-time reverse transcription polymerase chain reaction (RT-PCR method for the diagnosis of the Flavivirus genus. METHODS: We evaluated different commercial kits that use the SYBR Green system for real-time RT-PCR with a primer set that amplifies a fragment of the NS5 flavivirus gene. The specificity and sensitivity of the assay were tested using twelve flaviviruses and ribonucleic acid (RNA transcribed from the yellow fever virus. Additionally, this assay was evaluated using the sera of 410 patients from different regions of Brazil with acute febrile illness and a negative diagnosis for the dengue virus. RESULTS: The real-time RT-PCR amplified all flaviviruses tested at a melting temperature of 79.92 to 83.49°C. A detection limit of 100 copies per ml was determined for this assay. Surprisingly, we detected dengue virus in 4.1% (17/410 of samples from patients with febrile illness and a supposedly negative dengue infection diagnosis. The viral load in patients ranged from 2.1×107to 3.4×103copies per ml. CONCLUSIONS: The real-time RT-PCR method may be very useful for preliminary diagnoses in screenings, outbreaks, and other surveillance studies. Moreover, this assay can be easily applied to monitor viral activity and to measure viral load in pathogenesis studies.

  11. Identification of chloroquine resistance Pfcrt-K76T and determination of Pfmdr1-N86Y copy number by SYBR Green I qPCR

    Directory of Open Access Journals (Sweden)

    Addimas Tajebe

    2015-03-01

    Conclusions: The study showed high prevalence level and fixation of Pfcrt, 76T mutation after chloroquine withdrawal. The prevalence of Pfmdr1 copy number variant suggested that the presence of modulating factor for emergence of Plasmodium falciparum strains with higher copy numbers. However, the prevalence level was not statistically significant.

  12. A novel rapid genotyping technique for Collie eye anomaly: SYBR Green-based real-time polymerase chain reaction method applicable to blood and saliva specimens on Flinders Technology Associates filter paper.

    Science.gov (United States)

    Chang, Hye-Sook; Mizukami, Keijiro; Yabuki, Akira; Hossain, Mohammad A; Rahman, Mohammad M; Uddin, Mohammad M; Arai, Toshiro; Yamato, Osamu

    2010-09-01

    Collie eye anomaly (CEA) is a canine inherited ocular disease that shows a wide variety of manifestations and severity of clinical lesions. Recently, a CEA-associated mutation was reported, and a DNA test that uses conventional polymerase chain reaction (PCR) has now become available. The objective of the current study was to develop a novel rapid genotyping technique by using SYBR Green-based real-time PCR for future large-scale surveys as a key part in the strategy to eradicate CEA by selective breeding. First, a SYBR Green-based real-time PCR assay for genotyping of CEA was developed and evaluated by using purified DNA samples from normal, carrier, and affected Border Collies in which genotypes had previously been determined by conventional PCR. This real-time PCR assay demonstrated appropriate amplifications in all genotypes, and the results were consistent with those of conventional PCR. Second, the availability of Flinders Technology Associates filter paper (FTA card) as DNA templates for the real-time PCR assay was evaluated by using blood and saliva specimens to determine suitability for CEA screening. DNA-containing solution prepared from a disc of blood- or saliva-spotted FTA cards was available directly as templates for the real-time PCR assay when the volume of solution was 2.5% of the PCR mixture. In conclusion, SYBR Green-based real-time PCR combined with FTA cards is a rapid genotyping technique for CEA that can markedly shorten the overall time required for genotyping as well as simplify the sample preparation. Therefore, this newly developed technique suits large-scale screening in breeding populations of Collie-related breeds.

  13. MULTIPLEX SYBR® GREEN-REAL TIME PCR (qPCR ASSAY FOR THE DETECTION AND DIFFERENTIATION OF Bartonella henselae AND Bartonella clarridgeiae IN CATS

    Directory of Open Access Journals (Sweden)

    Rodrigo Staggemeier

    2014-04-01

    Full Text Available A novel SYBR® green-real time polymerase chain reaction (qPCR was developed to detect two Bartonella species, B. henselae and B. clarridgeiae, directly from blood samples. The test was used in blood samples obtained from cats living in animal shelters in Southern Brazil. Results were compared with those obtained by conventional PCR targeting Bartonella spp. Among the 47 samples analyzed, eight were positive using the conventional PCR and 12 were positive using qPCR. Importantly, the new qPCR detected the presence of both B. henselae and B. clarridgeiae in two samples. The results show that the qPCR described here may be a reliable tool for the screening and differentiation of two important Bartonella species.

  14. Real-time PCR based on SYBR-Green I fluorescence: An alternative to the TaqMan assay for a relative quantification of gene rearrangements, gene amplifications and micro gene deletions

    Directory of Open Access Journals (Sweden)

    Puisieux Alain

    2003-10-01

    Full Text Available Abstract Background Real-time PCR is increasingly being adopted for RNA quantification and genetic analysis. At present the most popular real-time PCR assay is based on the hybridisation of a dual-labelled probe to the PCR product, and the development of a signal by loss of fluorescence quenching as PCR degrades the probe. Though this so-called 'TaqMan' approach has proved easy to optimise in practice, the dual-labelled probes are relatively expensive. Results We have designed a new assay based on SYBR-Green I binding that is quick, reliable, easily optimised and compares well with the published assay. Here we demonstrate its general applicability by measuring copy number in three different genetic contexts; the quantification of a gene rearrangement (T-cell receptor excision circles (TREC in peripheral blood mononuclear cells; the detection and quantification of GLI, MYC-C and MYC-N gene amplification in cell lines and cancer biopsies; and detection of deletions in the OPA1 gene in dominant optic atrophy. Conclusion Our assay has important clinical applications, providing accurate diagnostic results in less time, from less biopsy material and at less cost than assays currently employed such as FISH or Southern blotting.

  15. SYBR green-based real-time reverse transcription-PCR for typing and subtyping of all hemagglutinin and neuraminidase genes of avian influenza viruses and comparison to standard serological subtyping tests

    Science.gov (United States)

    Tsukamoto, K.; Javier, P.C.; Shishido, M.; Noguchi, D.; Pearce, J.; Kang, H.-M.; Jeong, O.M.; Lee, Y.-J.; Nakanishi, K.; Ashizawa, T.

    2012-01-01

    Continuing outbreaks of H5N1 highly pathogenic (HP) avian influenza virus (AIV) infections of wild birds and poultry worldwide emphasize the need for global surveillance of wild birds. To support the future surveillance activities, we developed a SYBR green-based, real-time reverse transcriptase PCR (rRT-PCR) for detecting nucleoprotein (NP) genes and subtyping 16 hemagglutinin (HA) and 9 neuraminidase (NA) genes simultaneously. Primers were improved by focusing on Eurasian or North American lineage genes; the number of mixed-base positions per primer was set to five or fewer, and the concentration of each primer set was optimized empirically. Also, 30 cycles of amplification of 1:10 dilutions of cDNAs from cultured viruses effectively reduced minor cross- or nonspecific reactions. Under these conditions, 346 HA and 345 NA genes of 349 AIVs were detected, with average sensitivities of NP, HA, and NA genes of 10 1.5, 10 2.3, and 10 3.1 50% egg infective doses, respectively. Utility of rRT-PCR for subtyping AIVs was compared with that of current standard serological tests by using 104 recent migratory duck virus isolates. As a result, all HA genes and 99% of the NA genes were genetically subtyped, while only 45% of HA genes and 74% of NA genes were serologically subtyped. Additionally, direct subtyping of AIVs in fecal samples was possible by 40 cycles of amplification: approximately 70% of HA and NA genes of NP gene-positive samples were successfully subtyped. This validation study indicates that rRT-PCR with optimized primers and reaction conditions is a powerful tool for subtyping varied AIVs in clinical and cultured samples. Copyright ?? 2012, American Society for Microbiology. All Rights Reserved.

  16. Touch-down reverse transcriptase-PCR detection of IgV(H) rearrangement and Sybr-Green-based real-time RT-PCR quantitation of minimal residual disease in patients with chronic lymphocytic leukemia.

    Science.gov (United States)

    Peková, Sona; Marková, Jana; Pajer, Petr; Dvorák, Michal; Cetkovský, Petr; Schwarz, Jirí

    2005-01-01

    Patients with chronic lymphocytic leukemia (CLL) can relapse even after aggressive therapy and autografts. It is commonly assumed that to prevent relapse the level of minimal residual disease (MRD) should be as low as possible. To evaluate MRD, highly sensitive quantitative assays are needed. The aim of the study was to develop a robust and sensitive method for detection of the clonal immunoglobulin heavy-chain variable (IgV(H)) rearrangement in CLL and to introduce a highly sensitive and specific methodology for MRD monitoring in patients with CLL who undergo intensive treatment. As a prerequisite for MRD detection, touch-down reverse transcriptase (RT)-PCR using degenerate primers were used for the diagnostic identification of (H) gene rearrangement(s). For quantitative MRD detection in 18 patients, we employed a real-time RT-PCR assay (RQ-PCR) making use of patient-specific primers and the cost-saving Sybr-Green reporter dye (SG). For precise calibration of RQ-PCR, patient-specific IgV(H) sequences were cloned. Touch-down RT-PCR with degenerate primers allowed the successful detection of IgV(H) clonal rearrangement(s) in 252 of 257 (98.1%) diagnostic samples. Biallelic rearrangements were found in 27 of 252 (10.7%) cases. Degenerate primers used for the identification of clonal expansion at diagnosis were not sensitive enough for MRD detection. In contrast, our RQ-PCR assay using patient-specific primers and SG reached the sensitivity of 10(-)(6). We demonstrated MRD in each patient tested, including four of four patients in complete remission following autologous hematopoietic stem cell transplantation (HSCT) and three of three following allogeneic 'mini'-HSCT. Increments in MRD might herald relapse; aggressive chemotherapy could induce molecular remission. Our touch-down RT-PCR has higher efficiency to detect clonal IgV(H) rearrangements including the biallelic ones. MRD quantitation of IgV(H) expression using SG-based RQ-PCR represents a highly specific

  17. Duplex real-time PCR assay using SYBR Green to detect and quantify Malayan box turtle (Cuora amboinensis) materials in meatballs, burgers, frankfurters and traditional Chinese herbal jelly powder.

    Science.gov (United States)

    Asing; Ali, Eaqub; Hamid, Sharifah Bee Abd; Hossain, Motalib; Ahamad, Mohammad Nasir Uddin; Hossain, S M Azad; Naquiah, Nina; Zaidul, I S M

    2016-11-01

    The Malayan box turtle (Cuora amboinensis) (MBT) is a vulnerable and protected species widely used in exotic foods and traditional medicines. Currently available polymerase chain reaction (PCR) assays to identify MBT lack automation and involve long targets which break down in processed or denatured tissue. This SYBR Green duplex real-time PCR assay has addressed this research gap for the first time through the combination of 120- and 141-bp targets from MBT and eukaryotes for the quantitative detection of MBT DNA in food chain and herbal medicinal preparations. This authentication ensures better security through automation, internal control and short targets that were stable under the processing treatments of foods and medicines. A melting curve clearly demonstrated two peaks at 74.63 ± 0.22 and 78.40 ± 0.31°C for the MBT and eukaryotic products, respectively, under pure, admixed and commercial food matrices. Analysis of 125 reference samples reflected a target recovery of 93.25-153.00%, PCR efficiency of 99-100% and limit of detection of 0.001% under various matrices. The quantification limits were 0.00001, 0.00170 ± 0.00012, 0.00228 ± 0.00029, 0.00198 ± 0.00036 and 0.00191 ± 0.00043 ng DNA for the pure meat, binary mixtures, meatball, burger and frankfurter products, respectively. The assay was used to screen 100 commercial samples of traditional Chinese herbal jelly powder from eight different brands; 22% of them were found to be MBT-positive (5.37 ± 0.50-7.00 ± 0.34% w/w), which was reflected through the Ct values (26.37 ± 0.32-28.90 ± 0.42) and melting curves (74.63-78.65 ± 0.22°C) of the amplified MBT target (120 bp), confirming the speculation that MBT materials are widely used in Chinese herbal desserts, exotic dishes consumed with the hope of prolonging life and youth.

  18. Validation of kinetics similarity in qPCR

    Czech Academy of Sciences Publication Activity Database

    Bar, T.; Kubista, Mikael; Tichopád, Aleš

    2012-01-01

    Roč. 40, č. 4 (2012), s. 1395-1406 ISSN 0305-1048 R&D Projects: GA ČR GAP303/10/1338; GA ČR GA301/09/1752 Institutional research plan: CEZ:AV0Z50520701 Keywords : REAL-TIME PCR * POLYMERASE-CHAIN-REACTION * SYBR-GREEN-I Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 8.278, year: 2012

  19. Development of a multiplex qPCR in real time for quantification and differential diagnosis of Salmonella Gallinarum and Salmonella Pullorum.

    Science.gov (United States)

    Rubio, Marcela da Silva; Penha Filho, Rafael Antonio Casarin; Almeida, Adriana Maria de; Berchieri, Angelo

    2017-12-01

    Currently there are 2659 Salmonella serovars. The host-specific biovars Salmonella Pullorum and Salmonella Gallinarum cause systemic infections in food-producing and wild birds. Fast diagnosis is crucial to control the dissemination in avian environments. The present work describes the development of a multiplex qPCR in real time using a low-cost DNA dye (SYBr Green) to identify and quantify these biovars. Primers were chosen based on genomic regions of difference (RoD) and optimized to control dimers. Primers pSGP detect both host-specific biovars but not other serovars and pSG and pSP differentiate biovars. Three amplicons showed different melting temperatures (Tm), allowing differentiation. The pSGP amplicon (97 bp) showed Tm of 78°C for both biovars. The pSG amplicon (273 bp) showed a Tm of 86.2°C for S. Gallinarum and pSP amplicon (260 bp) dissociated at 84.8°C for S. Pullorum identification. The multiplex qPCR in real time showed high sensitivity and was capable of quantifying 10 8 -10 1 CFU of these biovars.

  20. A Java program for LRE-based real-time qPCR that enables large-scale absolute quantification.

    Science.gov (United States)

    Rutledge, Robert G

    2011-03-02

    Linear regression of efficiency (LRE) introduced a new paradigm for real-time qPCR that enables large-scale absolute quantification by eliminating the need for standard curves. Developed through the application of sigmoidal mathematics to SYBR Green I-based assays, target quantity is derived directly from fluorescence readings within the central region of an amplification profile. However, a major challenge of implementing LRE quantification is the labor intensive nature of the analysis. Utilizing the extensive resources that are available for developing Java-based software, the LRE Analyzer was written using the NetBeans IDE, and is built on top of the modular architecture and windowing system provided by the NetBeans Platform. This fully featured desktop application determines the number of target molecules within a sample with little or no intervention by the user, in addition to providing extensive database capabilities. MS Excel is used to import data, allowing LRE quantification to be conducted with any real-time PCR instrument that provides access to the raw fluorescence readings. An extensive help set also provides an in-depth introduction to LRE, in addition to guidelines on how to implement LRE quantification. The LRE Analyzer provides the automated analysis and data storage capabilities required by large-scale qPCR projects wanting to exploit the many advantages of absolute quantification. Foremost is the universal perspective afforded by absolute quantification, which among other attributes, provides the ability to directly compare quantitative data produced by different assays and/or instruments. Furthermore, absolute quantification has important implications for gene expression profiling in that it provides the foundation for comparing transcript quantities produced by any gene with any other gene, within and between samples.

  1. A Java program for LRE-based real-time qPCR that enables large-scale absolute quantification.

    Directory of Open Access Journals (Sweden)

    Robert G Rutledge

    Full Text Available BACKGROUND: Linear regression of efficiency (LRE introduced a new paradigm for real-time qPCR that enables large-scale absolute quantification by eliminating the need for standard curves. Developed through the application of sigmoidal mathematics to SYBR Green I-based assays, target quantity is derived directly from fluorescence readings within the central region of an amplification profile. However, a major challenge of implementing LRE quantification is the labor intensive nature of the analysis. FINDINGS: Utilizing the extensive resources that are available for developing Java-based software, the LRE Analyzer was written using the NetBeans IDE, and is built on top of the modular architecture and windowing system provided by the NetBeans Platform. This fully featured desktop application determines the number of target molecules within a sample with little or no intervention by the user, in addition to providing extensive database capabilities. MS Excel is used to import data, allowing LRE quantification to be conducted with any real-time PCR instrument that provides access to the raw fluorescence readings. An extensive help set also provides an in-depth introduction to LRE, in addition to guidelines on how to implement LRE quantification. CONCLUSIONS: The LRE Analyzer provides the automated analysis and data storage capabilities required by large-scale qPCR projects wanting to exploit the many advantages of absolute quantification. Foremost is the universal perspective afforded by absolute quantification, which among other attributes, provides the ability to directly compare quantitative data produced by different assays and/or instruments. Furthermore, absolute quantification has important implications for gene expression profiling in that it provides the foundation for comparing transcript quantities produced by any gene with any other gene, within and between samples.

  2. Evaluation of 16S rRNA qPCR for detection of Mycobacterium leprae DNA in nasal secretion and skin biopsy samples from multibacillary and paucibacillary leprosy cases.

    Science.gov (United States)

    Marques, Lívia Érika Carlos; Frota, Cristiane Cunha; Quetz, Josiane da Silva; Bindá, Alexandre Havt; Mota, Rosa Maria Salane; Pontes, Maria Araci de Andrade; Gonçalves, Heitor de Sá; Kendall, Carl; Kerr, Ligia Regina Franco Sansigolo

    2017-12-26

    Mycobacterium leprae bacilli are mainly transmitted by the dissemination of nasal aerosols from multibacillary (MB) patients to susceptible individuals through inhalation. The upper respiratory tract represents the main entry and exit routes of M. leprae. Therefore, this study aimed to evaluate the sensitivity and specificity of real-time quantitative polymerase chain reaction (qPCR) in detecting M. leprae in nasal secretion (NS) and skin biopsy (SB) samples from MB and paucibacillary (PB) cases. Fifty-four NS samples were obtained from leprosy patients at the Dona Libânia National Reference Centre for Sanitary Dermatology in Ceará, Brazil. Among them, 19 MB cases provided both NS and SB samples. Bacilloscopy index assays were conducted and qPCR amplification was performed using specific primers for M. leprae 16S rRNA gene, generating a 124-bp fragment. Primer specificity was verified by determining the amplicon melting temperature (T m  = 79.5 °C) and detection limit of qPCR was 20 fg of M. leprae DNA. Results were positive for 89.7 and 73.3% of NS samples from MB and PB cases, respectively. SB samples from MB patients were 100% positive. The number of bacilli detected in NS samples were 1.39 × 10 3 -8.02 × 10 5 , and in SB samples from MB patients were 1.87 × 10 3 -1.50 × 10 6 . Therefore, qPCR assays using SYBR Green targeting M. leprae 16S rRNA region can be employed in detecting M. leprae in nasal swabs from leprosy patients, validating this method for epidemiological studies aiming to identify healthy carriers among household contacts or within populations of an endemic area.

  3. Detection and quantification of Spongospora subterranea f. sp. subterranea in bait plants and potato fields in Colombia using QPCR

    International Nuclear Information System (INIS)

    Garcia Bastidas, Nevar; Morales, Juan Gonzalo; Gonzalez Jaimes, Paola; Gutierrez, Pablo Andres; Marin Montoya, Mauricio

    2013-01-01

    In recent years, potato crops (Solanum tuberosum, S. phureja) have been seriously affected by powdery scab; a disease caused by Spongospora subterranea f.sp. subterranea (Sss). In Colombia, asymptomatic detection of Sss has been achieved with bait plants, PCR of its regions and ELISA tests. Unfortunately, these techniques have low sensitivity and may require long processing times. In this work, quantitative real time PCR (qPCR) was tested for detection of Sss using different sets of primers. Primers SsTQF1-SsTQR1, Spon421F-Spon494R and SscolF-SscolR (designed in this study), were tested using SYBR green, while primers sponfsponr were tested using the Taqman probe sponp. Primers Spon421F-Spon494R was discarded due to lack of specificity. Standard curves were obtained from serial dilutions of Cystosori. the 20 N. benthamiana and potato bait plants evaluated tested positive for Sss using primers SsTQF1-SsTQR1 (Ct: 10.57-29.34) and Sscolf-SscolR (Ct: 14.39-34.08) and 19 samples were positive with primers SponF-SponR-SponP, with Ct values ranging between 15,63 and 38,93. Sss was detected in 17 out of 20 root samples from potato crops in la Union (Antioquia) using primers SscolF-SscolRt with an estimated concentration of 6470 to 1,39x10 1 0 cystosori/ mL. these results suggest high levels of sss in the potato fields from this region and recall the importance of strengthening seed-certification programs in Colombia.

  4. A comparative study of digital PCR and real-time qPCR for the detection and quantification of HPV mRNA in sentinel lymph nodes of cervical cancer patients.

    Science.gov (United States)

    Carow, Katrin; Read, Christina; Häfner, Norman; Runnebaum, Ingo B; Corner, Adam; Dürst, Matthias

    2017-10-30

    Qualitative analyses showed that the presence of HPV mRNA in sentinel lymph nodes of cervical cancer patients with pN0 status is associated with significantly decreased recurrence free survival. To further address the clinical potential of the strategy and to define prognostic threshold levels it is necessary to use a quantitative assay. Here, we compare two methods of quantification: digital PCR and standard quantitative PCR. Serial dilutions of 5 ng-5 pg RNA (≙ 500-0.5 cells) of the cervical cancer cell line SiHa were prepared in 5 µg RNA of the HPV-negative human keratinocyte cell line HaCaT. Clinical samples consisted of 10 sentinel lymph nodes with varying HPV transcript levels. Reverse transcription of total RNA (5 µg RNA each) was performed in 100 µl and cDNA aliquots were analyzed by qPCR and dPCR. Digital PCR was run in the RainDrop ® Digital PCR system (RainDance Technologies) using a probe-based detection of HPV E6/E7 cDNA PCR products with 11 µl template. qPCR was done using a Rotor Gene Q 5plex HRM (Qiagen) amplifying HPV E6/E7 cDNA in a SYBR Green format with 1 µl template. For the analysis of both, clinical samples and serial dilution samples, dPCR and qPCR showed comparable sensitivity. With regard to reproducibility, both methods differed considerably, especially for low template samples. Here, we found with qPCR a mean variation coefficient of 126% whereas dPCR enabled a significantly lower mean variation coefficient of 40% (p = 0.01). Generally, we saw with dPCR a substantial reduction of subsampling errors, which most likely reflects the large cDNA amounts available for analysis. Compared to real-time PCR, dPCR shows higher reliability. Thus, our HPV mRNA dPCR assay holds promise for the clinical evaluation of occult tumor cells in histologically tumor-free lymph nodes in future studies.

  5. Accuracy of qPCR for quantifying Leishmania kDNA in different skin layers of patients with American tegumentary leishmaniasis.

    Science.gov (United States)

    Sevilha-Santos, L; Dos Santos Júnior, A C M; Medeiros-Silva, V; Bergmann, J O; da Silva, E F; Segato, L F; Arabi, A Y M; de Paula, N A; Sampaio, R N R; Lima, B D; Gomes, C M

    2018-05-03

    Superficial swab sampling of American tegumentary leishmaniasis (ATL) lesions shows higher amounts of Leishmania than those from biopsy. Subcutaneous involvement is also important in ATL, but parasite quantification according to lesion depth has not been evaluated. We aim to present the best depth at which sampling should be performed for molecular exams of ATL. Patients with a clinical presentation compatible with ATL were allocated to ATL and control groups. Qualitative and quantitative qPCR assays were performed using SYBR Green and primers amplifying the kDNA minicircle of Leishmania spp. in different skin layers, including the epidermis, the superior dermis, the inferior dermis, and the hypodermis. Fifty-nine patients were included in this study, including 40 who had been diagnosed with ATL and 19 controls. The number of parasites was greater in samples of the epidermis and superior dermis (159.1 × 10 6 , range 4.0-781.7, and 75.4 × 10 6 , range 8.0-244.5, mean Leishmania parasite equivalents per μg of tissue DNA, respectively) than those in samples of the inferior dermis and hypodermis (54.6, range 8.0-256.6, and 16.8 × 10 6 , range 8.0-24.1, mean Leishmania parasite equivalents per μg of tissue DNA, respectively). The best diagnostic accuracy was achieved in the superior dermis (77.9%) and was significantly greater than that in the hypodermis (63.3%; p 0.039). We conclude that superficial sampling can retrieve a greater quantity of parasites. Future studies of the role of transepidermal elimination as a mechanism of host defence in ATL must be performed as there is a considerable quantity of Leishmania kDNA in the epidermis. Copyright © 2018 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  6. DETECCIÓN Y CUANTIFICACIÓN DE Spongospora subterranea f. sp. subterranea EN PLANTAS SEÑUELO Y CULTIVOS DE PAPA EN COLOMBIA MEDIANTE qPCR

    Directory of Open Access Journals (Sweden)

    Nevar Alirio García Bastidas

    2013-01-01

    Full Text Available La sarna polvosa de la papa (Solanum tuberosum , S. phureja causada por Spongospora subterranea f. sp. subterranea (Sss, es una de las enfermedades más limitantes de este cultivo. En Colombia, se han empleado diferentes métodos de detección asintomática de Sss, incluyendo bioensayos con plantas señuelo, PCR de ITS y pruebas de ELISA. Sin embargo, sus niveles de sensibilidad son bajos o requieren tiempos extensos. Una alternativa para complementar dichas herramientas es la PCR cuantitativa en tiempo real (qPCR. En este trabajo se evaluó dicha técnica utilizando los juegos de cebadores SsTQF1-SsTQR1; Spon421F-Spon494R y SscolF-SscolR (diseñados en este estudio, bajo la metodología de SYBR Green®; mientras que con Taqman® se evaluaron los cebadores SponF-SponR y la sonda SponP. Una vez determinada la funcionalidad de los cebadores, se descartó por inespecificidad, el par Spon421F-Spon494R; para los restantes se realizaron curvas estándar basadas en diluciones seriadas de quistosoros. Las pruebas de qPCR detectaron a Sss en las 20 muestras evaluadas de plantas señuelo de Nicotiana benthamiana y papa, utilizando los cebadores SsTQF1-SsTQR1 (Ct: 10,57- 29,34 y SscolF-SscolR (Ct: 14,39-34,08; mientras que 19 de las muestras fueron positivas con SponF-SponR-SponP (Ct: 15,63-38,93. A partir de 20 muestras de raíces de papa de cultivos de La Unión (Antioquia, Colombia, fue posible detectar el patógeno en 17 de ellas con SscolF-SscolR, estimándose una concentración de 6470 a 1,39 x 1010 quistorosos/mL. Estos resultados indican la ocurrencia de altos niveles de inóculo de Sss en esta región y enfatizan en la necesidad de fortalecer los programas de certificación de tubérculo-semilla en Colombia.

  7. green

    Directory of Open Access Journals (Sweden)

    Elena Grigoryeva

    2011-02-01

    Full Text Available The “green” topic follows the “youngsters”, which is quite natural for the Russian language.Traditionally these words put together sound slightly derogatory. However, “green” also means fresh, new and healthy.For Russia, and for Siberia in particular, “green” architecture does sound new and fresh. Forced by the anxious reality, we are addressing this topic intentionally. The ecological crisis, growing energy prices, water, air and food deficits… Alexander Rappaport, our regular author, writes: “ It has been tolerable until a certain time, but under transition to the global civilization, as the nature is destroyed, and swellings of megapolises expand incredibly fast, the size and the significance of all these problems may grow a hundredfold”.However, for this very severe Siberian reality the newness of “green” architecture may turn out to be well-forgotten old. A traditional Siberian house used to be built on principles of saving and environmental friendliness– one could not survive in Siberia otherwise.Probably, in our turbulent times, it is high time to fasten “green belts”. But we should keep from enthusiastic sticking of popular green labels or repainting of signboards into green color. We should avoid being drowned in paper formalities under “green” slogans. And we should prevent the Earth from turning into the planet “Kin-dza-dza”.

  8. SYBR safeTM efficiently replaces ethidium bromide in Aspergillus fumigatus gene disruption.

    Science.gov (United States)

    Canela, H M S; Takami, L A; Ferreira, M E S

    2017-02-08

    Invasive aspergillosis is a disease responsible for high mortality rates, caused mainly by Aspergillus fumigatus. The available drugs are limited and this disease continues to occur at an unacceptable frequency. Gene disruption is essential in the search for new drug targets. An efficient protocol for A. fumigatus gene disruption was described but it requires ethidium bromide, a genotoxic agent, for DNA staining. Therefore, the present study tested SYBR safe TM , a non-genotoxic DNA stain, in A. fumigatus gene disruption protocol. The chosen gene was cipC, which has already been disrupted successfully in our laboratory. A deletion cassette was constructed in Saccharomyces cerevisiae and used in A. fumigatus transformation. There was no statistical difference between the tested DNA stains. The success rate of S. cerevisiae transformation was 63.3% for ethidium bromide and 70% for SYBR safe TM . For A. fumigatus gene disruption, the success rate for ethidium bromide was 100 and 97% for SYBR safe TM . In conclusion, SYBR safe TM efficiently replaced ethidium bromide, making this dye a safe and efficient alternative for DNA staining in A. fumigatus gene disruption.

  9. A reliable and feasible qPCR strategy for titrating AAV vectors.

    Science.gov (United States)

    Wang, Feng; Cui, Xiuling; Wang, Mingxi; Xiao, Weidong; Xu, Ruian

    2013-07-05

    Previous studies have revealed that traditional real-time quantitative PCR (qPCR) underestimates adeno-associated virus (AAV) titer. Because the inverted terminal repeat (ITR) exists in all AAV vectors, the only remaining element from the wild genome could form special configurations to interfere with qPCR titration. To solve this problem, a modified and universal qPCR method was tested and established. In this work, there was a great variation in titration of ssAAV2-EGFP (Enhanced Green Fluorescence Protein) and scAAV2-EGFP genome by traditional qPCR. For ssAAV2-EGFP, the highest titer was found by using the targeting EGFP primers and the lowest titer was measured by those targeting bovine growth hormone polyA element (pBGH) primers. Experimental data were reverse for ssAAV2-EGFP and scAAV2-EGFP. Here we report an improved and universal SmaI qPCR method, based on cleaving all ITRs in AAV2 genome by SmaI with several advantages: (1) impact of all ITRs in ssAAV2 and scAAV2 was dismissed; (2) titers increased remarkably, up to 7-fold, especially for scAAV2; (3) the variation of titers was reduced when different primers were applied. A similar phenomenon was also observed in other ssAAV2 and scAAV2 products when the range of titration was at 3×107 to 7×109 V.G/µl in this study. This modified qPCR strategy can increase rAAV' titer and reduce titration variance, possibly become a universal method for titrating AAV vectors.

  10. Real-time PCR (qPCR) primer design using free online software.

    Science.gov (United States)

    Thornton, Brenda; Basu, Chhandak

    2011-01-01

    Real-time PCR (quantitative PCR or qPCR) has become the preferred method for validating results obtained from assays which measure gene expression profiles. The process uses reverse transcription polymerase chain reaction (RT-PCR), coupled with fluorescent chemistry, to measure variations in transcriptome levels between samples. The four most commonly used fluorescent chemistries are SYBR® Green dyes and TaqMan®, Molecular Beacon or Scorpion probes. SYBR® Green is very simple to use and cost efficient. As SYBR® Green dye binds to any double-stranded DNA product, its success depends greatly on proper primer design. Many types of online primer design software are available, which can be used free of charge to design desirable SYBR® Green-based qPCR primers. This laboratory exercise is intended for those who have a fundamental background in PCR. It addresses the basic fluorescent chemistries of real-time PCR, the basic rules and pitfalls of primer design, and provides a step-by-step protocol for designing SYBR® Green-based primers with free, online software. Copyright © 2010 Wiley Periodicals, Inc.

  11. Identification of Optimal Reference Genes for Normalization of qPCR Analysis during Pepper Fruit Development

    Directory of Open Access Journals (Sweden)

    Yuan Cheng

    2017-06-01

    Full Text Available Due to its high sensitivity and reproducibility, quantitative real-time PCR (qPCR is practiced as a useful research tool for targeted gene expression analysis. For qPCR operations, the normalization with suitable reference genes (RGs is a crucial step that eventually determines the reliability of the obtained results. Although pepper is considered an ideal model plant for the study of non-climacteric fruit development, at present no specific RG have been developed or validated for the qPCR analyses of pepper fruit. Therefore, this study aimed to identify stably expressed genes for their potential use as RGs in pepper fruit studies. Initially, a total of 35 putative RGs were selected by mining the pepper transcriptome data sets derived from the PGP (Pepper Genome Platform and PGD (Pepper Genome Database. Their expression stabilities were further measured in a set of pepper (Capsicum annuum L. var. 007e fruit samples, which represented four different fruit developmental stages (IM: Immature; MG: Mature green; B: Break; MR: Mature red using the qPCR analysis. Then, based on the qPCR results, three different statistical algorithms, namely geNorm, Normfinder, and boxplot, were chosen to evaluate the expression stabilities of these putative RGs. It should be noted that nine genes were proven to be qualified as RGs during pepper fruit development, namely CaREV05 (CA00g79660; CaREV08 (CA06g02180; CaREV09 (CA06g05650; CaREV16 (Capana12g002666; CaREV21 (Capana10g001439; CaREV23 (Capana05g000680; CaREV26 (Capana01g002973; CaREV27 (Capana11g000123; CaREV31 (Capana04g002411; and CaREV33 (Capana08g001826. Further analysis based on geNorm suggested that the application of the two most stably expressed genes (CaREV05 and CaREV08 would provide optimal transcript normalization in the qPCR experiments. Therefore, a new and comprehensive strategy for the identification of optimal RGs was developed. This strategy allowed for the effective normalization of the qPCR

  12. Efficacy of SYBR 14/propidium iodide viability stain for the amphibian chytrid fungus Batrachochytrium dendrobatidis.

    Science.gov (United States)

    Stockwell, M P; Clulow, J; Mahony, M J

    2010-01-25

    The amphibian chytrid fungus Batrachochytrium dendrobatidis is a recently described pathogen that has been implicated as a causal agent in the global decline in amphibians. Research into its biology and epidemiology has frequently involved in vitro experimentation. However, this research is currently limited by the inability to differentiate between viable and inviable zoospores. Stains are frequently used to determine cell viability, and this study tested a 2-colour fluorescence assay for the detection and quantification of viable B. dendrobatidis zoospores. The results show that the nucleic acid stains SYBR 14 and propidium iodide are effective in distinguishing live from dead zoospores, and a protocol has been optimized for their use. This viability assay provides an efficient and reliable tool that will have applications in B. dendrobatidis challenge and amphibian exposure experiments.

  13. Real-Time Detection and Identification of Chlamydophila Species in Veterinary Specimens by Using SYBR Green-Based PCR Assays

    DEFF Research Database (Denmark)

    Nordentoft, Steen; Kabell, Susanne; Pedersen, Karl

    2011-01-01

    of Chlamydiaceae and differentiate the most prevalent veterinary Chlamydophila species: Cp. psittaci, Cp. abortus, Cp. felis, and Cp. caviae. By adding bovine serum albumin to the master mixes, target DNA could be detected directly in crude lysates of enzymatically digested conjunctival or pharyngeal swabs...... or tissue specimens from heart, liver, and spleen without further purification. The assays were evaluated on veterinary specimens where all samples were screened using a family-specific PCR, and positive samples were further tested using species-specific PCRs. Cp. psittaci was detected in 47 birds, Cp...... with a highly sensitive family-specific PCR, we were able to screen for Chlamydiaceae in veterinary specimens and confirm the species in positive samples with additional PCR assays....

  14. Visualization of DNA-containing structurs in various species of Chlorophyta, Rhodophyta and Cyanophyta using SYBR green I dye

    Czech Academy of Sciences Publication Activity Database

    Vítová, Milada; Hendrychová, Jana; Cepák, Vladislav; Zachleder, Vilém

    2005-01-01

    Roč. 50, č. 4 (2005), s. 333-340 ISSN 0015-5632 R&D Projects: GA ČR GA204/02/1438; GA ČR GA204/03/1113; GA AV ČR KJB5020305 Institutional research plan: CEZ:AV0Z5020903; CEZ:AV0Z60050516 Keywords : dna-containing * chlorophyta * rhodophyta Subject RIV: EE - Microbiology, Virology Impact factor: 0.918, year: 2005

  15. A novel strategy for human papillomavirus detection and genotyping with SybrGreen and molecular beacon polymerase chain reaction

    NARCIS (Netherlands)

    Szuhai, K; Sandhaus, E; Kolkman-Uljee, SM; Lemaitre, M; Truffert, JC; Dirks, RW; Tanke, HJ; Fleuren, GJ; Schuuring, E; Raap, AK

    2001-01-01

    Human papillomaviruses (HPVs) play an important role in the pathogenesis of cervical cancer. For identification of the large number of different HPV types found in (pre)malignant lesions, a robust methodology is needed that combines general HPV detection with HPV genotyping. We have developed for

  16. Diagnosis of Barmah Forest virus infection by a nested real-time SYBR green RT-PCR assay.

    Directory of Open Access Journals (Sweden)

    Linda Hueston

    Full Text Available Barmah Forest virus (BFV is a mosquito borne (+ ssRNA alphavirus found only in Australia. It causes rash, myalgia and arthralgia in humans and is usually diagnosed serologically. We developed a real-time PCR assay to detect BFV in an effort to improve diagnosis early in the course of infection. The limit of detection was 16 genome equivalents with a specificity of 100%. Fifty five serum samples from BFV-infected patients were tested by the PCR. 52 of 53 antibody-positive samples were PCR negative. Two culture-positive (neutralizing antibody negative samples were positive on first round PCR, while one sample (IgM and neutralizing antibody strongly positive, IgG negative was positive on second round PCR, suggesting that viral RNA is detectable and transiently present in early infection. PCR can provide results faster than culture, is capable of high throughput and by sequencing the PCR product strain variants can be characterized.

  17. Molecular detection of Phytophthora ramorum by real-time PCR using Taqman, SYBR Green and molecular beacons with three genes

    Science.gov (United States)

    G.J. Bilodeau; C.A. Lévesque; A.W.A.M. De Cock; C. Duchaine; G. Kristjansson; R.C. Hamelin

    2006-01-01

    Sudden oak death, caused by Phytophthora ramorum, is a severe disease that can affect numerous species of trees and shrubs. This pathogen has been spread via nursery stock, and quarantine measures are currently in place to prevent further spread. Molecular assays have been developed to rapidly detect and identify P. ramorum, but...

  18. Diagnosis of Barmah Forest Virus Infection by a Nested Real-Time SYBR Green RT-PCR Assay

    OpenAIRE

    Hueston, Linda; Toi, Cheryl S.; Jeoffreys, Neisha; Sorrell, Tania; Gilbert, Gwendolyn

    2013-01-01

    Barmah Forest virus (BFV) is a mosquito borne (+) ssRNA alphavirus found only in Australia. It causes rash, myalgia and arthralgia in humans and is usually diagnosed serologically. We developed a real-time PCR assay to detect BFV in an effort to improve diagnosis early in the course of infection. The limit of detection was 16 genome equivalents with a specificity of 100%. Fifty five serum samples from BFV-infected patients were tested by the PCR. 52 of 53 antibody-positive samples were PCR ne...

  19. Comparison of Gull Feces-specific Assays Targeting the 16S rRNA Gene of Catellicoccus Marimammalium and Streptococcus spp.

    Science.gov (United States)

    Two novel gull-specific qPCR assays were developed using 16S rRNA gene sequences from gull fecal clone libraries: a SYBR-green-based assay targeting Streptococcus spp. (i.e., gull3) and a TaqMan qPCR assay targeting Catellicoccus marimammalium (i.e., gull4). The main objectives ...

  20. Cross-platform comparison of SYBR® Green real-time PCR with TaqMan PCR, microarrays and other gene expression measurement technologies evaluated in the MicroArray Quality Control (MAQC study

    Directory of Open Access Journals (Sweden)

    Dial Stacey L

    2008-07-01

    Full Text Available Abstract Background The MicroArray Quality Control (MAQC project evaluated the inter- and intra-platform reproducibility of seven microarray platforms and three quantitative gene expression assays in profiling the expression of two commercially available Reference RNA samples (Nat Biotechnol 24:1115-22, 2006. The tested microarrays were the platforms from Affymetrix, Agilent Technologies, Applied Biosystems, GE Healthcare, Illumina, Eppendorf and the National Cancer Institute, and quantitative gene expression assays included TaqMan® Gene Expression PCR Assay, Standardized (Sta RT-PCR™ and QuantiGene®. The data showed great consistency in gene expression measurements across different microarray platforms, different technologies and test sites. However, SYBR® Green real-time PCR, another common technique utilized by half of all real-time PCR users for gene expression measurement, was not addressed in the MAQC study. In the present study, we compared the performance of SYBR Green PCR with TaqMan PCR, microarrays and other quantitative technologies using the same two Reference RNA samples as the MAQC project. We assessed SYBR Green real-time PCR using commercially available RT2 Profiler™ PCR Arrays from SuperArray, containing primer pairs that have been experimentally validated to ensure gene-specificity and high amplification efficiency. Results The SYBR Green PCR Arrays exhibit good reproducibility among different users, PCR instruments and test sites. In addition, the SYBR Green PCR Arrays have the highest concordance with TaqMan PCR, and a high level of concordance with other quantitative methods and microarrays that were evaluated in this study in terms of fold-change correlation and overlap of lists of differentially expressed genes. Conclusion These data demonstrate that SYBR Green real-time PCR delivers highly comparable results in gene expression measurement with TaqMan PCR and other high-density microarrays.

  1. Metaphysical green

    DEFF Research Database (Denmark)

    Earon, Ofri

    2011-01-01

    to adapt to urban environment. It explores the potential of Sensation of Green in the city. The paper questions whether the Sensation of Green could introduce a new spectrum of greens, beside the real green. It develops the term of metaphysical green – does green have to be green or can it be only...

  2. Green(ing) infrastructure

    CSIR Research Space (South Africa)

    Van Wyk, Llewellyn V

    2014-03-01

    Full Text Available the generation of electricity from renewable sources such as wind, water and solar. Grey infrastructure – In the context of storm water management, grey infrastructure can be thought of as the hard, engineered systems to capture and convey runoff..., pumps, and treatment plants.  Green infrastructure reduces energy demand by reducing the need to collect and transport storm water to a suitable discharge location. In addition, green infrastructure such as green roofs, street trees and increased...

  3. Towards green loyalty: the influences of green perceived risk, green image, green trust and green satisfaction

    Science.gov (United States)

    Chrisjatmiko, K.

    2018-01-01

    The paper aims to present a comprehensive framework for the influences of green perceived risk, green image, green trust and green satisfaction to green loyalty. The paper also seeks to account explicitly for the differences in green perceived risk, green image, green trust, green satisfaction and green loyalty found among green products customers. Data were obtained from 155 green products customers. Structural equation modeling was used in order to test the proposed hypotheses. The findings show that green image, green trust and green satisfaction has positive effects to green loyalty. But green perceived risk has negative effects to green image, green trust and green satisfaction. However, green perceived risk, green image, green trust and green satisfaction also seems to be a good device to gain green products customers from competitors. The contributions of the paper are, firstly, a more complete framework of the influences of green perceived risk, green image, green trust and green satisfaction to green loyalty analyses simultaneously. Secondly, the study allows a direct comparison of the difference in green perceived risk, green image, green trust, green satisfaction and green loyalty between green products customers.

  4. HF183/BFDrev and HumM2 qPCR data

    Data.gov (United States)

    U.S. Environmental Protection Agency — Concentration estimates for HF183/BFDrev and HumM2 qPCR genetic markers in raw sewage collected from 54 geographic locations across the United States. This dataset...

  5. QPCR: Application for real-time PCR data management and analysis

    Directory of Open Access Journals (Sweden)

    Eichhorn Heiko

    2009-08-01

    Full Text Available Abstract Background Since its introduction quantitative real-time polymerase chain reaction (qPCR has become the standard method for quantification of gene expression. Its high sensitivity, large dynamic range, and accuracy led to the development of numerous applications with an increasing number of samples to be analyzed. Data analysis consists of a number of steps, which have to be carried out in several different applications. Currently, no single tool is available which incorporates storage, management, and multiple methods covering the complete analysis pipeline. Results QPCR is a versatile web-based Java application that allows to store, manage, and analyze data from relative quantification qPCR experiments. It comprises a parser to import generated data from qPCR instruments and includes a variety of analysis methods to calculate cycle-threshold and amplification efficiency values. The analysis pipeline includes technical and biological replicate handling, incorporation of sample or gene specific efficiency, normalization using single or multiple reference genes, inter-run calibration, and fold change calculation. Moreover, the application supports assessment of error propagation throughout all analysis steps and allows conducting statistical tests on biological replicates. Results can be visualized in customizable charts and exported for further investigation. Conclusion We have developed a web-based system designed to enhance and facilitate the analysis of qPCR experiments. It covers the complete analysis workflow combining parsing, analysis, and generation of charts into one single application. The system is freely available at http://genome.tugraz.at/QPCR

  6. A survey of tools for the analysis of quantitative PCR (qPCR) data.

    Science.gov (United States)

    Pabinger, Stephan; Rödiger, Stefan; Kriegner, Albert; Vierlinger, Klemens; Weinhäusel, Andreas

    2014-09-01

    Real-time quantitative polymerase-chain-reaction (qPCR) is a standard technique in most laboratories used for various applications in basic research. Analysis of qPCR data is a crucial part of the entire experiment, which has led to the development of a plethora of methods. The released tools either cover specific parts of the workflow or provide complete analysis solutions. Here, we surveyed 27 open-access software packages and tools for the analysis of qPCR data. The survey includes 8 Microsoft Windows, 5 web-based, 9 R-based and 5 tools from other platforms. Reviewed packages and tools support the analysis of different qPCR applications, such as RNA quantification, DNA methylation, genotyping, identification of copy number variations, and digital PCR. We report an overview of the functionality, features and specific requirements of the individual software tools, such as data exchange formats, availability of a graphical user interface, included procedures for graphical data presentation, and offered statistical methods. In addition, we provide an overview about quantification strategies, and report various applications of qPCR. Our comprehensive survey showed that most tools use their own file format and only a fraction of the currently existing tools support the standardized data exchange format RDML. To allow a more streamlined and comparable analysis of qPCR data, more vendors and tools need to adapt the standardized format to encourage the exchange of data between instrument software, analysis tools, and researchers.

  7. Quantitative Real-Time PCR using the Thermo Scientific Solaris qPCR Assay

    Science.gov (United States)

    Ogrean, Christy; Jackson, Ben; Covino, James

    2010-01-01

    The Solaris qPCR Gene Expression Assay is a novel type of primer/probe set, designed to simplify the qPCR process while maintaining the sensitivity and accuracy of the assay. These primer/probe sets are pre-designed to >98% of the human and mouse genomes and feature significant improvements from previously available technologies. These improvements were made possible by virtue of a novel design algorithm, developed by Thermo Scientific bioinformatics experts. Several convenient features have been incorporated into the Solaris qPCR Assay to streamline the process of performing quantitative real-time PCR. First, the protocol is similar to commonly employed alternatives, so the methods used during qPCR are likely to be familiar. Second, the master mix is blue, which makes setting the qPCR reactions easier to track. Third, the thermal cycling conditions are the same for all assays (genes), making it possible to run many samples at a time and reducing the potential for error. Finally, the probe and primer sequence information are provided, simplifying the publication process. Here, we demonstrate how to obtain the appropriate Solaris reagents using the GENEius product search feature found on the ordering web site (www.thermo.com/solaris) and how to use the Solaris reagents for performing qPCR using the standard curve method. PMID:20567213

  8. A common base method for analysis of qPCR data and the application of simple blocking in qPCR experiments.

    Science.gov (United States)

    Ganger, Michael T; Dietz, Geoffrey D; Ewing, Sarah J

    2017-12-01

    qPCR has established itself as the technique of choice for the quantification of gene expression. Procedures for conducting qPCR have received significant attention; however, more rigorous approaches to the statistical analysis of qPCR data are needed. Here we develop a mathematical model, termed the Common Base Method, for analysis of qPCR data based on threshold cycle values (C q ) and efficiencies of reactions (E). The Common Base Method keeps all calculations in the logscale as long as possible by working with log 10 (E) ∙ C q , which we call the efficiency-weighted C q value; subsequent statistical analyses are then applied in the logscale. We show how efficiency-weighted C q values may be analyzed using a simple paired or unpaired experimental design and develop blocking methods to help reduce unexplained variation. The Common Base Method has several advantages. It allows for the incorporation of well-specific efficiencies and multiple reference genes. The method does not necessitate the pairing of samples that must be performed using traditional analysis methods in order to calculate relative expression ratios. Our method is also simple enough to be implemented in any spreadsheet or statistical software without additional scripts or proprietary components.

  9. Technical aspects and recommendations for single-cell qPCR.

    Science.gov (United States)

    Ståhlberg, Anders; Kubista, Mikael

    2018-02-01

    Single cells are basic physiological and biological units that can function individually as well as in groups in tissues and organs. It is central to identify, characterize and profile single cells at molecular level to be able to distinguish different kinds, to understand their functions and determine how they interact with each other. During the last decade several technologies for single-cell profiling have been developed and used in various applications, revealing many novel findings. Quantitative PCR (qPCR) is one of the most developed methods for single-cell profiling that can be used to interrogate several analytes, including DNA, RNA and protein. Single-cell qPCR has the potential to become routine methodology but the technique is still challenging, as it involves several experimental steps and few molecules are handled. Here, we discuss technical aspects and provide recommendation for single-cell qPCR analysis. The workflow includes experimental design, sample preparation, single-cell collection, direct lysis, reverse transcription, preamplification, qPCR and data analysis. Detailed reporting and sharing of experimental details and data will promote further development and make validation studies possible. Efforts aiming to standardize single-cell qPCR open up means to move single-cell analysis from specialized research settings to standard research laboratories. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Legionella detection by culture and qPCR: Comparing apples and oranges.

    Science.gov (United States)

    Whiley, Harriet; Taylor, Michael

    2016-01-01

    Legionella spp. are the causative agent of Legionnaire's disease and an opportunistic pathogen of significant public health concern. Identification and quantification from environmental sources is crucial for identifying outbreak origins and providing sufficient information for risk assessment and disease prevention. Currently there are a range of methods for Legionella spp. quantification from environmental sources, but the two most widely used and accepted are culture and real-time polymerase chain reaction (qPCR). This paper provides a review of these two methods and outlines their advantages and limitations. Studies from the last 10 years which have concurrently used culture and qPCR to quantify Legionella spp. from environmental sources have been compiled. 26/28 studies detected Legionella at a higher rate using qPCR compared to culture, whilst only one study detected equivalent levels of Legionella spp. using both qPCR and culture. Aggregating the environmental samples from all 28 studies, 2856/3967 (72%) tested positive for the presence of Legionella spp. using qPCR and 1331/3967 (34%) using culture. The lack of correlation between methods highlights the need to develop an acceptable standardized method for quantification that is sufficient for risk assessment and management of this human pathogen.

  11. Detection of foodborne pathogens by qPCR: A practical approach for food industry applications

    Directory of Open Access Journals (Sweden)

    María-José Chapela

    2015-12-01

    Full Text Available Microbiological analysis of food is an integrated part of microbial safety management in the food chain. Monitoring and controlling foodborne pathogens are traditionally carried out by conventional microbiological methods based on culture-dependent approaches in control laboratories and private companies. However, polymerase chain reaction (PCR has revolutionized microbiological analysis allowing detection of pathogenic microorganisms in food, without the necessity of classical isolation and identification. However, at present, PCR and quantitative polymerase chain reaction (qPCR are essential analytical tools for researchers working in the field of foodborne pathogens. This manuscript reviews recently described qPCR methods applied for foodborne bacteria detection, serving as economical, safe, and reliable alternatives for application in the food industry and control laboratories. Multiplex qPCR, which allows the simultaneous detection of more than one pathogen in one single reaction, saving considerable effort, time, and money, is emphasized in the article.

  12. Cooverexpression of EpCAM and c-myc genes in malignant breast ...

    Indian Academy of Sciences (India)

    SAMIRA SADEGHI

    Samira Sadeghi et al. domain protein2), and the transcription factor Lef1 that is ... antibody approved in. European Market in 2009, to reduce the rate of metastasis in .... Real-Time PCR. System and Maxima SYBR Green/ROX qPCR Master Mix.

  13. Green Tourism

    OpenAIRE

    Hasan, Ali

    2014-01-01

    Green tourism is defined as environmentally friendly tourism activities with various focuses and meanings. In a broad term, green tourism is about being an environmentally friendly tourist or providing environmentally friendly tourist services. The green tourism concept would be highly appealing to tourism enterprises and operators owing to increasing governmental pressure to improve environmental performance by adopting effective and tangible environmental management techniques. Green to...

  14. Metaphysical green

    OpenAIRE

    Earon, Ofri

    2011-01-01

    “Sensation of Green is about the mental process like touching, seeing, hearing, or smelling, resulting from the immediate stimulation of landscape forms, plants, trees, wind and water. Sensation of Green triggers a feeling of scale, cheerfulness, calmness and peace. The spatial performance of Sensation of Green is created by a physical interaction between the language of space and the language of nature” The notion of Sensation of Green was developed through a previous study ‘Learning from th...

  15. Longitudinal monitoring of bottlenose dolphin leukocyte cytokine mRNA responsiveness by qPCR

    Science.gov (United States)

    Both veterinarians caring for bottlenose dolphins (Tursiops truncatus) in managed populations and researchers monitoring wild populations use blood-based diagnostics to monitor bottlenose dolphin health. Quantitative PCR (qPCR) can be used to assess cytokine expression patterns of peripheral blood m...

  16. Real-Time PCR (qPCR) Primer Design Using Free Online Software

    Science.gov (United States)

    Thornton, Brenda; Basu, Chhandak

    2011-01-01

    Real-time PCR (quantitative PCR or qPCR) has become the preferred method for validating results obtained from assays which measure gene expression profiles. The process uses reverse transcription polymerase chain reaction (RT-PCR), coupled with fluorescent chemistry, to measure variations in transcriptome levels between samples. The four most…

  17. The need for transparency and good practices in the qPCR literature

    DEFF Research Database (Denmark)

    Bustin, Stephen A; Benes, Vladimir; Garson, Jeremy

    2013-01-01

    Two surveys of over 1,700 publications whose authors use quantitative real-time PCR (qPCR) reveal a lack of transparent and comprehensive reporting of essential technical information. Reporting standards are significantly improved in publications that cite the Minimum Information for Publication...

  18. A survey of tools for the analysis of quantitative PCR (qPCR data

    Directory of Open Access Journals (Sweden)

    Stephan Pabinger

    2014-09-01

    Our comprehensive survey showed that most tools use their own file format and only a fraction of the currently existing tools support the standardized data exchange format RDML. To allow a more streamlined and comparable analysis of qPCR data, more vendors and tools need to adapt the standardized format to encourage the exchange of data between instrument software, analysis tools, and researchers.

  19. Identification of Listeria monocytogenes on Green Mussels (Perna viridis and Cockle Shell (Anadara granosa

    Directory of Open Access Journals (Sweden)

    Winiati Puji Rahayu

    2016-12-01

    Full Text Available Green mussel (Perna viridis and cockle shell (Anadara granosa are one of many sources of animalprotein which is many cultivated in Indonesia because their price is relatively affordable. This study wasconducted to identify the presence of Listeria monocytogenes in 27 samples of green mussels and 3 samplesof cockle shells using real-time Polymerase Chain Reaction (real-time PCR and biochemical methods. Thetarget gene for amplification in real-time PCR was an hlyA gene because this gene was a determinant ofvirulence genes that produce listeriolysin O. Primers used in this study were forward primer DG69 (GTGCCG GGT AAA AGA CCA TA and reverse primer DG74 (CGC CAC TGA GAT ACT AT and fluorescencesignals indicator using SYBR Green I. The results of analysis using real-time PCR were negative Listeriamonocytogenes in all samples, while using biochemical methods there was one of 30 samples contaminatedby Listeria welshimeri.

  20. Metaphysical green

    DEFF Research Database (Denmark)

    Earon, Ofri

    2011-01-01

    example is a tiny Danish summer house from 1918 . The second example is ‘House before House’ , in Tokyo. The third example is a prefabricated house ‘CHU’ . The analysis evaluates the characteristics of diverse tones of green – from green image to green sensation. The analysis is based on the original...... of Sensation of Green is created by a physical interaction between the language of space and the language of nature” The notion of Sensation of Green was developed through a previous study ‘Learning from the Summer House’ investigating the unique architectural characteristics of the Danish summer houses...... the Sensation of Green? Three existing examples are agents to this discussion. The first example is a Danish summer house. The other two are international urban examples. While the summer house articulates the original meaning of Sensation of Green, the urban examples illustrate its urban context. The first...

  1. Why the need for qPCR publication guidelines?--The case for MIQE.

    Science.gov (United States)

    Bustin, Stephen A

    2010-04-01

    The polymerase chain reaction (PCR) has matured from a labour- and time-intensive, low throughput qualitative gel-based technique to an easily automated, rapid, high throughput quantitative technology. Real-time quantitative PCR (qPCR) has become the benchmark technology for the detection and quantification of nucleic acids in a research, diagnostic, forensic and biotechnology setting. However, ill-assorted pre-assay conditions, poor assay design and inappropriate data analysis methodologies have resulted in the recurrent publication of data that are at best inconsistent and at worst irrelevant and even misleading. Furthermore, there is a lamentable lack of transparency of reporting, with the "Materials and Methods" sections of many publications, especially those with high impact factors, not fit for the purpose of evaluating the quality of any reported qPCR data. This poses a challenge to the integrity of the scientific literature, with serious consequences not just for basic research, but potentially calamitous implications for drug development and disease monitoring. These issues are being addressed by a set of guidelines that propose a minimum standard for the provision of information for qPCR experiments ("MIQE"). MIQE aims to restructure to-day's free-for-all qPCR methods into a more consistent format that will encourage detailed auditing of experimental detail, data analysis and reporting principles. General implementation of these guidelines is an important requisite for the maturing of qPCR into a robust, accurate and reliable nucleic acid quantification technology. Copyright 2009 Elsevier Inc. All rights reserved.

  2. Quantification of Paratrichodorus allius in DNA extracted from soil using TaqMan probe and SYBR green real-time PCR assays

    Science.gov (United States)

    The ectoparasitic stubby root nematode Paratrichodorus allius transmits Tobacco rattle virus, which causes corky ringspot disease resulting in significant economic losses in the potato industry. This study developed a diagnostic method for direct quantification of P. allius from soil DNA using a Taq...

  3. Identification by real-time PCR with SYBR Green of Leishmania spp. and Serratia marcescens in canine 'sterile' cutaneous nodular lesions.

    Science.gov (United States)

    Cornegliani, Luisa; Corona, Antonio; Vercelli, Antonella; Roccabianca, Paola

    2015-06-01

    Noninfectious, non-neoplastic, nodular to diffuse, so-called 'sterile' granulomatous/pyogranulomatous skin lesions (SGPSLs) are infrequently identified in dogs and may represent a diagnostic challenge. Their correct identification is based on history, histopathology and absence of intralesional foreign bodies and micro-organisms. The aim of this study was to investigate the presence of Leishmania spp., Mycobacterium spp., Serratia marcescens and Nocardia spp. by real-time PCR in canine nodular skin lesions histologically diagnosed as putatively sterile. Formalin-fixed skin biopsies were collected from 40 dogs. All samples were associated with an SGPSL diagnosis characterized by multifocal, nodular to diffuse, periadnexal and perifollicular pyogranulomas/granulomas. Neither micro-organisms nor foreign bodies were detected with haematoxylin and eosin staining, under polarized light. Further analyses included periodic acid Schiff, Ziehl-Neelsen, Fite Faraco, Giemsa and Gram histochemical stains; anti-Bacillus Calmette-Guérin (BCG) and Leishmania spp. immunohistochemistry; and real-time PCR analysis for Leishmania spp., Mycobacterium spp., S. marcescens and Nocardia spp. Special stains and BCG/immunohistochemistry were negative in all samples. Real-time PCR was positive for Leishmania spp. in four of 40 biopsies and for S. marcescens in two of 40 samples. Real-time PCR for Mycobacterium spp. and Nocardia spp. was negative. No correlation between real-time PCR positivity and a specific histological pattern was identified. Leishmania spp. have been previously identified as possible agents of certain SGPSLs, while the involvement of S. marcescens has not been investigated previously. According to our findings, Serratia spp. should be included in the list of agents possibly associated with a subgroup of granulomatous/pyogranulomatous skin lesions in dogs. © 2015 ESVD and ACVD.

  4. Improvement of High-throughput Genotype Analysis After Implementation of a Dual-curve Sybr Green I-based Quantification and Normalization Procedure

    Science.gov (United States)

    The ability to rapidly screen a large number of individuals is the key to any successful plant breeding program. One of the primary bottlenecks in high throughput screening is the preparation of DNA samples, particularly the quantification and normalization of samples for downstream processing. A ...

  5. Relative quantification and detection of different types of infectious bursal disease virus in bursa of Fabricius and cloacal swabs using real time RT-PCR SYBR green technology

    DEFF Research Database (Denmark)

    Li, Yiping; Handberg, K.J.; Kabell, Susanne

    2007-01-01

    or F52/70 inoculation were detected as virus positive at day I post inoculation (p.i.). The D78 viral load peaked at day 4 and day 8 p.i., while the DK01 and F52/70 viral load showed relatively high levels at day 2 p.i. In cloacal swabs, viruses detectable were at day 2 p.i. for DK01 and F52/70, day 8...

  6. Green Chemistry

    Energy Technology Data Exchange (ETDEWEB)

    Collison, Melanie

    2011-05-15

    Green chemistry is the science of chemistry used in a way that will not use or create hazardous substances. Dr. Rui Resendes is working in this field at GreenCentre Canada, an offshoot of PARTEQ Innovations in Kingston, Ontario. GreenCentre's preliminary findings suggest their licensed product {sup S}witchable Solutions{sup ,} featuring 3 classes of solvents and a surfactant, may be useful in bitumen oil sands extraction.

  7. Green roofs

    CSIR Research Space (South Africa)

    Van Wyk, Llewellyn V

    2010-04-01

    Full Text Available , beetles and spiders); and the number of birds that nest in vegetated roofs (including kestrels, swallows, and wagtails). Objective The primary objective of a green roof is to create a living habitat in an otherwise barren environment, hence the use... the negative environmental impacts including plant and insect specie loss. Thus at a philosophical level green roofs support the notion “replace what you displace”. Key ecological issues that can be addressed through green roofs include: Negative effects...

  8. Green thunderstorms

    Science.gov (United States)

    Gallagher, Frank Woolsey, III

    Many people around the world have observed green light apparently emanating from severe thunderstorms, but until recently there has been no scientific study of the phenomenon. Green thunderstorms have been observed from time to time in association with deep convection or severe weather events. Some skeptics who have not personally observed a green thunderstorm suggest that they are some kind of illusion. The existence of green thunderstorms has been objectively demonstrated by recording spectra of light from thunderstorms using a handheld spectrophotometer. During the spring and summer of 1995 and the spring of 1996 numerous storms were observed and spectra of the light emanating from these storms were recorded. Observations were made both at the ground and aboard research aircraft. Furthermore, time series of spectra were recorded as the observed color of some storms changed from dark blue to a bluish-green. Several hypotheses have been advanced to explain the occurrence of green light in connection with severe storms. Fankhauser gave some observational support to the belief that green light from thunderstorms is possible and believed that the source of the light is from the blue sky penetrating thin regions in the clouds. Fraser believes that light from the setting sun, in combination with the process of scattering by atmospheric molecules, creates the green light associated with severe weather and the thunderstorm acts only as a black backdrop. Unfortunately, no cloud illuminated by the sun is black and the green airlight produced by the Fraser theory is in reality overwhelmed by light reflected by the cloud. Often the unusual coloration has been attributed to hail or to reflection of light from foliage on the ground. The quantitative measurements made during the observation period fail to support these assumptions. We have observed thunderstorms to be green over ground that was not green and we have observed blue thunderstorms over ground that was green

  9. Behaviorally Green

    DEFF Research Database (Denmark)

    Sunstein, Cass; Reisch, Lucia A.

    2016-01-01

    of suggestion, inertia, and loss aversion. If well-chosen, green defaults are likely to have large effects in reducing the economic and environmental harms associated with various products and activities. Such defaults may or may not be more expensive to consumers. In deciding whether to establish green...

  10. Green Tea

    Science.gov (United States)

    ... and cancer. Green tea is consumed as a beverage. It is also sold in liquid extracts, capsules, and tablets and is sometimes used in topical products (intended to be applied to the skin). How Much Do We Know? Although many studies have been done on green tea and its ...

  11. Development and optimization of an efficient qPCR system for olive authentication in edible oils.

    Science.gov (United States)

    Alonso-Rebollo, Alba; Ramos-Gómez, Sonia; Busto, María D; Ortega, Natividad

    2017-10-01

    The applicability of qPCR in olive-oil authentication depends on the DNA obtained from the oils and the amplification primers. Therefore, four olive-specific amplification systems based on the trnL gene were designed (A-, B-, C- and D-trnL systems). The qPCR conditions, primer concentration and annealing temperature, were optimized. The systems were tested for efficiency and sensitivity to select the most suitable for olive oil authentication. The selected system (D-trnL) demonstrated specificity toward olive in contrast to other oleaginous species (canola, soybean, sunflower, maize, peanut and coconut) and showed high sensitivity in a broad linear dynamic range (LOD and LOQ: 500ng - 0.0625pg). This qPCR system enabled detection, with high sensitivity and specificity, of olive DNA isolated from oils processed in different ways, establishing it as an efficient method for the authentication of olive oil regardless of its category. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Green consumerism

    DEFF Research Database (Denmark)

    de Groot, Judith I.M.; Schuitema, Geertje; Garson, Carrie Lee

    and biospheric values influence the importance of such ‘green’ product characteristics on purchasing intentions. In two within-subjects full-factorial experimental studies (N = 100 and N = 107), we found that purchase intentions of products were only steered by green characteristics if prices were low...... and the brand was familiar. Green product characteristics did not influence purchase intentions at all when these proself product characteristics were not fulfilled (i.e., high prices and unfamiliar brands). The importance of proself and green product characteristics on purchasing intentions was also......Our presentation will focus on the influence of product characteristics and values on green consumerism. Although generally a majority of consumers support the idea of purchasing green products, we argue, based on social dilemma theory, that proself product characteristics and egoistic...

  13. Green lights

    DEFF Research Database (Denmark)

    Fisker, Peter Kielberg

    This study investigates the effect of drought on economic activity globally using remote sensing data. In particular, predicted variation in greenness is correlated with changes in the density of artificial light observed at night on a grid of 0.25 degree latitude-longitude pixels. I define drought...... as greenness estimated by lagged variation in monthly rainfall and temperature. This definition of drought performs well in identifying self-reported drought events since 2000 compared with measures of drought that do not take greenness into account, and the subsequent analysis indicates that predicted...... variation in greenness is positively associated with year-on-year changes in luminosity: If a unit of observation experiences a predicted variation in greenness that lies 1 standard deviation below the global mean, on average 1.5 - 2.5 light pixels out of 900 are extinguished that year. Finally, an attempt...

  14. Sampling and Pooling Methods for Capturing Herd Level Antibiotic Resistance in Swine Feces using qPCR and CFU Approaches

    DEFF Research Database (Denmark)

    Schmidt, Gunilla Veslemøy; Mellerup, Anders; Christiansen, Lasse Engbo

    2015-01-01

    The aim of this article was to define the sampling level and method combination that captures antibiotic resistance at pig herd level utilizing qPCR antibiotic resistance gene quantification and culture-based quantification of antibiotic resistant coliform indicator bacteria. Fourteen qPCR assays...... for commonly detected antibiotic resistance genes were developed, and used to quantify antibiotic resistance genes in total DNA from swine fecal samples that were obtained using different sampling and pooling methods. In parallel, the number of antibiotic resistant coliform indicator bacteria was determined...... in the same swine fecal samples. The results showed that the qPCR assays were capable of detecting differences in antibiotic resistance levels in individual animals that the coliform bacteria colony forming units (CFU) could not. Also, the qPCR assays more accurately quantified antibiotic resistance genes...

  15. Construction of Genetically Engineered Streptococcus gordonii Strains to Provide Control in QPCR Assays for Assessing Microbiological Quality in Recreational Water.

    Science.gov (United States)

    Quantitative PCR (QPCR) methods for beach monitoring by estimating abundance of Enterococcus spp. in recreational waters use internal, positive controls which address only the amplification of target DNA. In this study two internal, positive controls were developed to control for...

  16. Green Engineering

    Science.gov (United States)

    Green Engineering is the design, commercialization and use of processes and products that are feasible and economical while reducing the generation of pollution at the source and minimizing the risk to human health and the environment.

  17. Green Roofs

    Energy Technology Data Exchange (ETDEWEB)

    None

    2004-08-01

    A New Technology Demonstration Publication Green roofs can improve the energy performance of federal buildings, help manage stormwater, reduce airborne emissions, and mitigate the effects of urban heat islands.

  18. Going Green

    Centers for Disease Control (CDC) Podcasts

    This podcast is for a general audience and provides information on how to recycle, re-use, and restore. It also covers the benefits of “Going Green" on the environment, health, and social interaction.

  19. Green lasers

    DEFF Research Database (Denmark)

    Jensen, Ole Bjarlin

    2010-01-01

    Well over a dozen papers at this year's Photonics West meeting in San Francisco boasted improvements in harmonic generation to produce visible laser beams, most of them in the green spectral range......Well over a dozen papers at this year's Photonics West meeting in San Francisco boasted improvements in harmonic generation to produce visible laser beams, most of them in the green spectral range...

  20. Green Nudging

    OpenAIRE

    Evans, Nicholas; Eickers, Stephanie; Geene, Leonie; Todorovic, Marijana; Villmow, Annika; Forschungsstelle für Umweltpolitik (FFU), Freie Universität Berlin

    2018-01-01

    Traditional environmental policy instruments have not always proven successful in fostering environmentally friendly behaviour. The question remains: how can policymakers tackle the attitude-behaviour gap when it comes to pro-environmental choices and sustainable lifestyles? One solution that has emerged is green nudging, a new and potentially promising policy tool born of behavioural economics and experimental psychology. This paper contributes to the current discussion surrounding green nud...

  1. qPCR detection of Mycobacterium leprae in biopsies and slit skin smear of different leprosy clinical forms

    Directory of Open Access Journals (Sweden)

    Michelle de Campos Soriani Azevedo

    2017-01-01

    Full Text Available Leprosy, whose etiological agent is Mycobacterium leprae, is a chronic infectious disease that mainly affects the skin and peripheral nervous system. The diagnosis of leprosy is based on clinical evaluation, whereas histopathological analysis and bacilloscopy are complementary diagnostic tools. Quantitative PCR (qPCR, a current useful tool for diagnosis of infectious diseases, has been used to detect several pathogens including Mycobacterium leprae. The validation of this technique in a robust set of samples comprising the different clinical forms of leprosy is still necessary. Thus, in this study samples from 126 skin biopsies (collected from patients on all clinical forms and reactional states of leprosy and 25 slit skin smear of leprosy patients were comparatively analyzed by qPCR (performed with primers for the RLEP region of M. leprae DNA and routine bacilloscopy performed in histological sections or in slit skin smear. Considering clinical diagnostic as the gold standard, 84.9% of the leprosy patients were qPCR positive in skin biopsies, resulting in 84.92% sensitivity, with 84.92 and 61.22% positive (PPV and negative (NPV predictive values, respectively. Concerning bacilloscopy of histological sections (BI/H, the sensitivity was 80.15% and the PPV and NPV were 80.15 and 44.44%, respectively. The concordance between qPCR and BI/H was 87.30%. Regarding the slit skin smear, 84% of the samples tested positive in the qPCR. Additionally, qPCR showed 100% specificity, since all samples from different mycobacteria, from healthy individuals, and from other granulomatous diseases presented negative results. In conclusion, the qPCR technique for detection of M. leprae using RLEP primers proved to be specific and sensitive, and qPCR can be used as a complementary test to diagnose leprosy irrespective of the clinical form of disease.

  2. qPCR detection of Mycobacterium leprae in biopsies and slit skin smear of different leprosy clinical forms.

    Science.gov (United States)

    Azevedo, Michelle de Campos Soriani; Ramuno, Natália Mortari; Fachin, Luciana Raquel Vincenzi; Tassa, Mônica; Rosa, Patrícia Sammarco; Belone, Andrea de Faria Fernandes; Diório, Suzana Madeira; Soares, Cleverson Teixeira; Garlet, Gustavo Pompermaier; Trombone, Ana Paula Favaro

    Leprosy, whose etiological agent is Mycobacterium leprae, is a chronic infectious disease that mainly affects the skin and peripheral nervous system. The diagnosis of leprosy is based on clinical evaluation, whereas histopathological analysis and bacilloscopy are complementary diagnostic tools. Quantitative PCR (qPCR), a current useful tool for diagnosis of infectious diseases, has been used to detect several pathogens including Mycobacterium leprae. The validation of this technique in a robust set of samples comprising the different clinical forms of leprosy is still necessary. Thus, in this study samples from 126 skin biopsies (collected from patients on all clinical forms and reactional states of leprosy) and 25 slit skin smear of leprosy patients were comparatively analyzed by qPCR (performed with primers for the RLEP region of M. leprae DNA) and routine bacilloscopy performed in histological sections or in slit skin smear. Considering clinical diagnostic as the gold standard, 84.9% of the leprosy patients were qPCR positive in skin biopsies, resulting in 84.92% sensitivity, with 84.92 and 61.22% positive (PPV) and negative (NPV) predictive values, respectively. Concerning bacilloscopy of histological sections (BI/H), the sensitivity was 80.15% and the PPV and NPV were 80.15 and 44.44%, respectively. The concordance between qPCR and BI/H was 87.30%. Regarding the slit skin smear, 84% of the samples tested positive in the qPCR. Additionally, qPCR showed 100% specificity, since all samples from different mycobacteria, from healthy individuals, and from other granulomatous diseases presented negative results. In conclusion, the qPCR technique for detection of M. leprae using RLEP primers proved to be specific and sensitive, and qPCR can be used as a complementary test to diagnose leprosy irrespective of the clinical form of disease. Copyright © 2016 Sociedade Brasileira de Infectologia. Published by Elsevier Editora Ltda. All rights reserved.

  3. Sources of Blood Meals of Sylvatic Triatoma guasayana near Zurima, Bolivia, Assayed with qPCR and 12S Cloning

    Science.gov (United States)

    Lucero, David E.; Ribera, Wilma; Pizarro, Juan Carlos; Plaza, Carlos; Gordon, Levi W.; Peña, Reynaldo; Morrissey, Leslie A.; Rizzo, Donna M.; Stevens, Lori

    2014-01-01

    Background In this study we compared the utility of two molecular biology techniques, cloning of the mitochondrial 12S ribosomal RNA gene and hydrolysis probe-based qPCR, to identify blood meal sources of sylvatic Chagas disease insect vectors collected with live-bait mouse traps (also known as Noireau traps). Fourteen T. guasayana were collected from six georeferenced trap locations in the Andean highlands of the department of Chuquisaca, Bolivia. Methodology/Principal Findings We detected four blood meals sources with the cloning assay: seven samples were positive for human (Homo sapiens), five for chicken (Gallus gallus) and unicolored blackbird (Agelasticus cyanopus), and one for opossum (Monodelphis domestica). Using the qPCR assay we detected chicken (13 vectors), and human (14 vectors) blood meals as well as an additional blood meal source, Canis sp. (4 vectors). Conclusions/Significance We show that cloning of 12S PCR products, which avoids bias associated with developing primers based on a priori knowledge, detected blood meal sources not previously considered and that species-specific qPCR is more sensitive. All samples identified as positive for a specific blood meal source by the cloning assay were also positive by qPCR. However, not all samples positive by qPCR were positive by cloning. We show the power of combining the cloning assay with the highly sensitive hydrolysis probe-based qPCR assay provides a more complete picture of blood meal sources for insect disease vectors. PMID:25474154

  4. Sources of blood meals of sylvatic Triatoma guasayana near Zurima, Bolivia, assayed with qPCR and 12S cloning.

    Directory of Open Access Journals (Sweden)

    David E Lucero

    2014-12-01

    Full Text Available In this study we compared the utility of two molecular biology techniques, cloning of the mitochondrial 12S ribosomal RNA gene and hydrolysis probe-based qPCR, to identify blood meal sources of sylvatic Chagas disease insect vectors collected with live-bait mouse traps (also known as Noireau traps. Fourteen T. guasayana were collected from six georeferenced trap locations in the Andean highlands of the department of Chuquisaca, Bolivia.We detected four blood meals sources with the cloning assay: seven samples were positive for human (Homo sapiens, five for chicken (Gallus gallus and unicolored blackbird (Agelasticus cyanopus, and one for opossum (Monodelphis domestica. Using the qPCR assay we detected chicken (13 vectors, and human (14 vectors blood meals as well as an additional blood meal source, Canis sp. (4 vectors.We show that cloning of 12S PCR products, which avoids bias associated with developing primers based on a priori knowledge, detected blood meal sources not previously considered and that species-specific qPCR is more sensitive. All samples identified as positive for a specific blood meal source by the cloning assay were also positive by qPCR. However, not all samples positive by qPCR were positive by cloning. We show the power of combining the cloning assay with the highly sensitive hydrolysis probe-based qPCR assay provides a more complete picture of blood meal sources for insect disease vectors.

  5. Amplification of nonspecific products in quantitative polymerase chain reactions (qPCR

    Directory of Open Access Journals (Sweden)

    Adrián Ruiz-Villalba

    2017-12-01

    Full Text Available Quantitative PCR allows the precise measurement of DNA concentrations and is generally considered to be straightforward and trouble free. However, a survey with 93 validated assays for genes in the Wnt-pathway showed that the amplification of nonspecific products occurs frequently and is unrelated to Cq or PCR efficiency values. Titration experiments showed that the occurrence of low and high melting temperature artifacts was shown to be determined by annealing temperature, primer concentration and cDNA input. To explore the range of input variations that occur in the normal use of the Cre assay these conditions were mimicked in a complete two-way design of template −plasmid DNA- and non-template −mouse cDNA- concentrations. These experiments showed that the frequency of the amplification of the correct product and the artifact, as well as the valid quantification of the correct product, depended on the concentration of the non-template cDNA. This finding questions the interpretation of dilution series in which template as well as non-template concentrations are simultaneously decreasing. Repetition of this cDNA concentration experiment with other templates revealed that exact reproduction qPCR experiments was affected by the time it takes to complete the pipetting of a qPCR plate. Long bench times were observed to lead to significantly more artifacts. However, the measurement of artifact-associated fluorescence can be avoided by inclusion of a small heating step after the elongation phase in the amplification protocol. Taken together, this trouble-shooting journey showed that reliability and reproducibility of qPCR experiments not only depends on standardization and reporting of the biochemistry and technical aspects but also on hitherto neglected factors as sample dilution and waiting times in the laboratory work flow. Keywords: RT-qPCR, Melting curve analysis, Reaction parameters, Artifacts

  6. Identification of Listeria monocytogenes on Green Mussels and Cockle Shell

    Directory of Open Access Journals (Sweden)

    Winiati Puji Rahayu

    2017-02-01

    Full Text Available AbstractGreen mussel (Perna viridis and cockle shell (Anadara granosa are one of many sources of animal protein which is many cultivated in Indonesia because their price is relatively affordable. This study was conducted to identify the presence of Listeria monocytogenes in 27 samples of green mussels and 3 samples of cockle shells using real-time Polymerase Chain Reaction (real-time PCR and biochemical methods. The target gene for amplification in real-time PCR was an hlyA gene because this gene was a determinant of virulence genes that produce listeriolysin O. Primers used in this study were forward primer DG69 (GTG CCG GGT AAA AGA CCA TA and reverse primer DG74 (CGC CAC TGA GAT ACT AT and fluorescence signals indicator using SYBR Green I. The results of analysis using real-time PCR were negative Listeria monocytogenes in all samples, while using biochemical methods there was one of 30 samples contaminated by Listeria welshimeri.

  7. Pre-amplification in the context of high-throughput qPCR gene expression experiment

    Czech Academy of Sciences Publication Activity Database

    Korenková, Vlasta; Scott, J.; Novosadová, Vendula; Jindřichová, Marie; Langerová, Lucie; Švec, David; Šídová, Monika; Sjoback, R.

    2015-01-01

    Roč. 16, č. 5 (2015) ISSN 1471-2199 R&D Projects: GA ČR(CZ) GAP304/12/1585; GA ČR(CZ) GA15-08239S; GA ČR GA13-02154S; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:86652036 Keywords : High-throughput qPCR * Gene expression * Exponential pre-amplification Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.500, year: 2015

  8. Development of a qPCR method to rapidly assess the function of NKT cells.

    Science.gov (United States)

    Sohn, Silke; Tiper, Irina; Japp, Emily; Sun, Wenji; Tkaczuk, Katherine; Webb, Tonya J

    2014-05-01

    NKT cells comprise a rare, but important subset of T cells which account for ~0.2% of the total circulating T cell population. NKT cells are known to have anti-tumor functions and rapidly produce high levels of cytokines following activation. Several clinical trials have sought to exploit the effector functions of NKT cells. While some studies have shown promise, NKT cells are approximately 50% lower in cancer patients compared to healthy donors of the same age and gender, thus limiting their therapeutic efficacy. These studies indicate that baseline levels of activation should be assessed before initiating an NKT cell based immunotherapeutic strategy. The goal of this study was to develop a sensitive method to rapidly assess NKT cell function. We utilized artificial antigen presenting cells in combination with qPCR in order to determine NKT cell function in peripheral blood mononuclear cells from healthy donors and breast cancer patients. We found that NKT cell activation can be detected by qPCR, but not by ELISA, in healthy donors as well as in breast cancer patients following four hour stimulation. This method utilizing CD1d-expressing aAPCs will enhance our knowledge of NKT cell biology and could potentially be used as a novel tool in adoptive immunotherapeutic strategies. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Is qPCR a Reliable Indicator of Cyanotoxin Risk in Freshwater?

    Science.gov (United States)

    Pacheco, Ana Beatriz F.; Guedes, Iame A.; Azevedo, Sandra M.F.O.

    2016-01-01

    The wide distribution of cyanobacteria in aquatic environments leads to the risk of water contamination by cyanotoxins, which generate environmental and public health issues. Measurements of cell densities or pigment contents allow both the early detection of cellular growth and bloom monitoring, but these methods are not sufficiently accurate to predict actual cyanobacterial risk. To quantify cyanotoxins, analytical methods are considered the gold standards, but they are laborious, expensive, time-consuming and available in a limited number of laboratories. In cyanobacterial species with toxic potential, cyanotoxin production is restricted to some strains, and blooms can contain varying proportions of both toxic and non-toxic cells, which are morphologically indistinguishable. The sequencing of cyanobacterial genomes led to the description of gene clusters responsible for cyanotoxin production, which paved the way for the use of these genes as targets for PCR and then quantitative PCR (qPCR). Thus, the quantification of cyanotoxin genes appeared as a new method for estimating the potential toxicity of blooms. This raises a question concerning whether qPCR-based methods would be a reliable indicator of toxin concentration in the environment. Here, we review studies that report the parallel detection of microcystin genes and microcystin concentrations in natural populations and also a smaller number of studies dedicated to cylindrospermopsin and saxitoxin. We discuss the possible issues associated with the contradictory findings reported to date, present methodological limitations and consider the use of qPCR as an indicator of cyanotoxin risk. PMID:27338471

  10. Is qPCR a Reliable Indicator of Cyanotoxin Risk in Freshwater?

    Directory of Open Access Journals (Sweden)

    Ana Beatriz F. Pacheco

    2016-06-01

    Full Text Available The wide distribution of cyanobacteria in aquatic environments leads to the risk of water contamination by cyanotoxins, which generate environmental and public health issues. Measurements of cell densities or pigment contents allow both the early detection of cellular growth and bloom monitoring, but these methods are not sufficiently accurate to predict actual cyanobacterial risk. To quantify cyanotoxins, analytical methods are considered the gold standards, but they are laborious, expensive, time-consuming and available in a limited number of laboratories. In cyanobacterial species with toxic potential, cyanotoxin production is restricted to some strains, and blooms can contain varying proportions of both toxic and non-toxic cells, which are morphologically indistinguishable. The sequencing of cyanobacterial genomes led to the description of gene clusters responsible for cyanotoxin production, which paved the way for the use of these genes as targets for PCR and then quantitative PCR (qPCR. Thus, the quantification of cyanotoxin genes appeared as a new method for estimating the potential toxicity of blooms. This raises a question concerning whether qPCR-based methods would be a reliable indicator of toxin concentration in the environment. Here, we review studies that report the parallel detection of microcystin genes and microcystin concentrations in natural populations and also a smaller number of studies dedicated to cylindrospermopsin and saxitoxin. We discuss the possible issues associated with the contradictory findings reported to date, present methodological limitations and consider the use of qPCR as an indicator of cyanotoxin risk.

  11. Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs.

    Directory of Open Access Journals (Sweden)

    Genevieve Housman

    2015-11-01

    Full Text Available Zoonotic pathogens that cause leprosy (Mycobacterium leprae and tuberculosis (Mycobacterium tuberculosis complex, MTBC continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were specific and sensitive to pathogen detection, while more traditional ELISAs were only specific. These assays were then employed in a case study to detect M. leprae as well as MTBC in wild marmosets. All marmosets were negative for M. leprae DNA, but 14 were positive for the mycobacterial rpoB gene assay. Targeted capture and sequencing of rpoB and other MTBC genes validated the presence of mycobacterial DNA in these samples and revealed that qPCR is useful for identifying mycobacterial-infected animal hosts.

  12. Rapid quantification of plant-powdery mildew interactions by qPCR and conidiospore counts.

    Science.gov (United States)

    Weßling, Ralf; Panstruga, Ralph

    2012-08-31

    The powdery mildew disease represents a valuable patho-system to study the interaction between plant hosts and obligate biotrophic fungal pathogens. Numerous discoveries have been made on the basis of the quantitative evaluation of plant-powdery mildew interactions, especially in the context of hyper-susceptible and/or resistant plant mutants. However, the presently available methods to score the pathogenic success of powdery mildew fungi are laborious and thus not well suited for medium- to high-throughput analysis. Here we present two new protocols that allow the rapid quantitative assessment of powdery mildew disease development. One procedure depends on quantitative polymerase chain reaction (qPCR)-based evaluation of fungal biomass, while the other relies on the quantification of fungal conidiospores. We validated both techniques using the powdery mildew pathogen Golovinomyces orontii on a set of hyper-susceptible and resistant Arabidopsis thaliana mutants and found that both cover a wide dynamic range of one to two (qPCR) and four to five (quantification of conidia) orders of magnitude, respectively. The two approaches yield reproducible results and are easy to perform without specialized equipment. The qPCR and spore count assays rapidly and reproducibly quantify powdery mildew pathogenesis. Our methods are performed at later stages of infection and discern mutant phenotypes accurately. The assays therefore complement currently used procedures of powdery mildew quantification and can overcome some of their limitations. In addition, they can easily be adapted to other plant-powdery mildew patho-systems.

  13. Single-cell qPCR on dispersed primary pituitary cells -an optimized protocol

    Directory of Open Access Journals (Sweden)

    Haug Trude M

    2010-11-01

    Full Text Available Abstract Background The incidence of false positives is a potential problem in single-cell PCR experiments. This paper describes an optimized protocol for single-cell qPCR measurements in primary pituitary cell cultures following patch-clamp recordings. Two different cell harvesting methods were assessed using both the GH4 prolactin producing cell line from rat, and primary cell culture from fish pituitaries. Results Harvesting whole cells followed by cell lysis and qPCR performed satisfactory on the GH4 cell line. However, harvesting of whole cells from primary pituitary cultures regularly produced false positives, probably due to RNA leakage from cells ruptured during the dispersion of the pituitary cells. To reduce RNA contamination affecting the results, we optimized the conditions by harvesting only the cytosol through a patch pipette, subsequent to electrophysiological experiments. Two important factors proved crucial for reliable harvesting. First, silanizing the patch pipette glass prevented foreign extracellular RNA from attaching to charged residues on the glass surface. Second, substituting the commonly used perforating antibiotic amphotericin B with β-escin allowed efficient cytosol harvest without loosing the giga seal. Importantly, the two harvesting protocols revealed no difference in RNA isolation efficiency. Conclusion Depending on the cell type and preparation, validation of the harvesting technique is extremely important as contaminations may give false positives. Here we present an optimized protocol allowing secure harvesting of RNA from single cells in primary pituitary cell culture following perforated whole cell patch clamp experiments.

  14. Green banking

    Directory of Open Access Journals (Sweden)

    Maja Drobnjaković

    2013-06-01

    Full Text Available There is an urgent need to march towards “low - carbon economy”. Global challenges of diminishing fossil fuel reserves, climate change, environmental management and finite natural resources serving an expanding world population - these reasons mean that urgent action is required to transition to solutions which minimize environmental impact and are sustainable. We are at the start of the low - carbon revolution and those that have started on their low - carbon journey already are seeing benefits such as new markets and customers, improved economic, social and environmental performance, and reduced bills and risks. Green investment banks offer alternative financial services: green car loans, energy efficiency mortgages, alternative energy venture capital, eco - savings deposits and green credit cards. These items represent innovative financial products.

  15. Miltenberger blood group typing by real-time polymerase chain reaction (qPCR) melting curve analysis in Thai population.

    Science.gov (United States)

    Vongsakulyanon, A; Kitpoka, P; Kunakorn, M; Srikhirin, T

    2015-12-01

    To develop reliable and convenient methods for Miltenberger (Mi(a) ) blood group typing. To apply real-time polymerase chain reaction (qPCR) melting curve analysis to Mi(a) blood group typing. The Mi(a) blood group is the collective set of glycophorin hybrids in the MNS blood group system. Mi(a+) blood is common among East Asians and is also found in the Thai population. Incompatible Mi(a) blood transfusions pose the risk of life-threatening haemolysis; therefore, Mi(a) blood group typing is necessary in ethnicities where the Mi(a) blood group is prevalent. One hundred and forty-three blood samples from Thai blood donors were used in the study. The samples included 50 Mi(a+) samples and 93 Mi(a-) samples, which were defined by serology. The samples were typed by Mi(a) typing qPCR, and 50 Mi(a+) samples were sequenced to identify the Mi(a) subtypes. Mi(a) subtyping qPCR was performed to define GP.Mur. Both Mi(a) typing and Mi(a) subtyping were tested on a conventional PCR platform. The results of Mi(a) typing qPCR were all concordant with serology. Sequencing of the 50 Mi(a+) samples revealed 47 GP.Mur samples and 3 GP.Hop or Bun samples. Mi(a) subtyping qPCR was the supplementary test used to further define GP.Mur from other Mi(a) subtypes. Both Mi(a) typing and Mi(a) subtyping performed well using a conventional PCR platform. Mi(a) typing qPCR correctly identified Mi(a) blood groups in a Thai population with the feasibility of Mi(a) subtype discrimination, and Mi(a) subtyping qPCR was able to further define GP.Mur from other Mi(a) subtypes. © 2015 British Blood Transfusion Society.

  16. Comparison of nested PCR and qPCR for the detection and quantitation of BoHV6 DNA.

    Science.gov (United States)

    Kubiś, Piotr; Materniak, Magdalena; Kuźmak, Jacek

    2013-12-01

    Nested PCR and qPCR (quantitative PCR) tests based on glycoprotein B (gB) gene were designed for detecting Bovine herpesvirus 6 (BoHV6) in bovine whole blood samples and wild ruminant blood clots (deer and roe-deer). This virus, commonly known as BLHV (bovine lymphotropic herpesvirus) belongs to the Herpesviridae family, subfamily Gammaherpesvirinae and Macavirus genus. DNA isolated from 92 dairy cow blood samples and 69 wild ruminant clots were examined for the presence of BoHV6 using nested PCR and qPCR tests. Viral DNA was detected by using nested PCR in 59 out of 92 bovine blood samples (64.1%), and by qPCR in 68 out of 92 bovine blood samples (73.9%), but none out of 69 DNA samples isolated from wild ruminant blood clots, was positive in both assays. The specificity of nested PCR and qPCR was confirmed by using BoHV1, BoHV4, BoHV6, BFV, BIV, and BLV DNA. The sensitivity of nested PCR and qPCR was determined using a serially 10-fold diluted vector pCR2.1HgB (2 × 10(0)-2 × 10(6)copies/reaction). In this testing, qPCR was more sensitive than the nested PCR, detecting two copies of BoHV6 whilst the limit of detection for nested PCR was 20 copies. In all qPCR assays, the coefficients of determination (R(2)) ranged between 0.990 and 0.999, and the calculated amplification efficiencies (Eff%) within the range of 89.7-106.9. The intra- and inter-assay CV (coefficient of variation) values did not exceed 4%. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. Green times

    International Nuclear Information System (INIS)

    Hasenclever, W.D.; Hasenclever, C.

    1982-01-01

    The authors, founding members of the ''Green Party'' have in mind to make a very personal contribution to a better understanding of the present political situation which, although it seems to have reached a deadlock, still offers positive chances and prospects. New approaches in policy are mentioned which may help to overcome the present state of resignation of many adolescents and adults. Among other things, they describe themselves setting out for new pathways, the ''Greens'' in Parliament, prospect for the future, opportunities of the ecologically oriented economic policy. Finally, they call upon the reader to think and develop further under the motto ''What we all can do''. (HSCH) [de

  18. Q-PCR Based Culture-Independent Enumeration and Detection of Enterobacter: An Emerging Environmental Human Pathogen in Riverine Systems and Potable Water

    Science.gov (United States)

    Patel, Chandra B.; Shanker, Rishi; Gupta, Vijai K.; Upadhyay, Ram S.

    2016-01-01

    The availability of safe and pristine water is a global challenge when large numbers of natural and anthropogenic water resources are being depleted with faster rate. The remaining water resources are severely contaminated with various kinds of contaminants including microorganisms. Enterobacter is one of the fecal coliform bacteria of family Enterobacteriaceae. Enterobacter was earlier used as an indicator bacterium along with other fecal Coliforms namely Escherichia coli, Citrobacter, and Klebsiella, but it is now known to cause various diseases in human beings. In this study, we have collected 55 samples from potable water and riverine system and proved their presence using their conserved sequences of 16S rRNA and 23S rRNA genes with the help of SYBR green real-time PCR, which showed very high specificity for the detection of Enterobacter. The Enterobacter counts in potable water were found to 1290 ± 32.89 to 1460 ± 39.42 cfu/100 ml. The Enterobacter levels in surface water were 1.76 × 104 ± 492, 1.33 × 104 ± 334, 1.15 × 104 ± 308, 2.56 × 104 ± 802, 2.89 × 104 ± 962, 8.16 × 104 ± 3443 cfu/100 ml; the levels of Enterobacter contamination associated with hydrophytes were 4.80 × 104 ± 1804, 3.48 × 104 ± 856, 8.50 × 104 ± 2074, 8.09 × 104 ± 1724, 6.30 × 104 ± 1738, 3.68 × 104 ± 949 cfu/10 g and the Enterobacter counts in sediments of the river, were 2.36 × 104 ± 703, 1.98 × 104 ± 530, 9.92 × 104 ± 3839, 6.80 × 104 ± 2230, 8.76 × 104 ± 3066 and 2.34 × 104 ± 732 cfu/10 g at the sampling Site #1, Site #2, Site #3, Site #4, Site #5, and Site #6, respectively. The assay could be used for the regular monitoring of potable water and other water reservoirs to check waterborne outbreaks. PMID:26925044

  19. Going Green

    Science.gov (United States)

    Witkowsky, Kathy

    2009-01-01

    Going green saves money and can even make money. Sustainable practices promote better health, less absenteeism, and more productivity. They also attract students, who are paying increasing attention to schools' environmental policies. Beyond being the smart thing to do, administrators at the University of Washington say repeatedly, it's the right…

  20. Buying Green

    Science.gov (United States)

    Layng, T. V. Joe

    2010-01-01

    In "Buying Green," Joe Layng recognizes that, like all choices we make, our decisions as consumers are more likely to be influenced by their short-term consequences for us as individuals (price, quality) than they are by their long-term consequences for society (environmental impact). He believes that the equation can be tilted in favor of greener…

  1. Green pioneers.

    Science.gov (United States)

    Trueland, Jennifer

    The government has set tough targets for the NHS in England to reduce its carbon footprint. In this article, nurses and managers at Nottinghamshire Healthcare NHS Trust explain how a programme of 'greening' initiatives - including a trial of electric cars for community staff - have slashed the trust's CO2 output.

  2. Automatically Green

    DEFF Research Database (Denmark)

    Sunstein, Cass R.; Reisch, Lucia

    2014-01-01

    reasons include the power of suggestion; inertia and procrastination; and loss aversion. If well-chosen, green defaults are likely to have large effects in reducing the economic and environmental harms associated with various products and activities. Such defaults may or may not be more expensive...

  3. Automatically Green

    DEFF Research Database (Denmark)

    Sunstein, Cass R.; Reisch, Lucia

    reasons include the power of suggestion; inertia and procrastination; and loss aversion. If well-chosen, green defaults are likely to have large effects in reducing the economic and environmental harms associated with various products and activities. Such defaults may or may not be more expensive...

  4. Going Green

    Centers for Disease Control (CDC) Podcasts

    2008-04-18

    This podcast is for a general audience and provides information on how to recycle, re-use, and restore. It also covers the benefits of “Going Green" on the environment, health, and social interaction.  Created: 4/18/2008 by National Center for Environmental Health (NCEH), ATSDR.   Date Released: 5/8/2008.

  5. Relationship and variation of qPCR and culturable enterococci estimates in ambient surface waters are predictable

    Science.gov (United States)

    Whitman, Richard L.; Ge, Zhongfu; Nevers, Meredith B.; Boehm, Alexandria B.; Chern, Eunice C.; Haugland, Richard A.; Lukasik, Ashley M.; Molina, Marirosa; Przybyla-Kelly, Kasia; Shively, Dawn A.; White, Emily M.; Zepp, Richard G.; Byappanahalli, Muruleedhara N.

    2010-01-01

    The quantitative polymerase chain reaction (qPCR) method provides rapid estimates of fecal indicator bacteria densities that have been indicated to be useful in the assessment of water quality. Primarily because this method provides faster results than standard culture-based methods, the U.S. Environmental Protection Agency is currently considering its use as a basis for revised ambient water quality criteria. In anticipation of this possibility, we sought to examine the relationship between qPCR-based and culture-based estimates of enterococci in surface waters. Using data from several research groups, we compared enterococci estimates by the two methods in water samples collected from 37 sites across the United States. A consistent linear pattern in the relationship between cell equivalents (CCE), based on the qPCR method, and colony-forming units (CFU), based on the traditional culturable method, was significant (P 10CFU > 2.0/100 mL) while uncertainty increases at lower CFU values. It was further noted that the relative error in replicated qPCR estimates was generally higher than that in replicated culture counts even at relatively high target levels, suggesting a greater need for replicated analyses in the qPCR method to reduce relative error. Further studies evaluating the relationship between culture and qPCR should take into account analytical uncertainty as well as potential differences in results of these methods that may arise from sample variability, different sources of pollution, and environmental factors.

  6. Comparison of ELISA, nested PCR and sequencing and a novel qPCR for detection of Giardia isolates from Jordan.

    Science.gov (United States)

    Hijjawi, Nawal; Yang, Rongchang; Hatmal, Ma'mon; Yassin, Yasmeen; Mharib, Taghrid; Mukbel, Rami; Mahmoud, Sameer Alhaj; Al-Shudifat, Abdel-Ellah; Ryan, Una

    2018-02-01

    Little is known about the prevalence of Giardia duodenalis in human patients in Jordan and all previous studies have used direct microscopy, which lacks sensitivity. The present study developed a novel quantitative PCR (qPCR) assay at the β-giardin (bg) locus and evaluated its use as a frontline test for the diagnosis of giardiasis in comparison with a commercially available ELISA using nested PCR and sequencing of the glutamate dehydrogenase (gdh) locus (gdh nPCR) as the gold standard. A total of 96 human faecal samples were collected from 96 patients suffering from diarrhoea from 5 regions of Jordan and were screened using the ELISA and qPCR. The analytical specificity of the bg qPCR assay revealed no cross-reactions with other genera and detected all the Giardia isolates tested. Analytical sensitivity was 1 Giardia cyst per μl of DNA extract. The overall prevalence of Giardia was 64.6%. The clinical sensitivity and specificity of the bg qPCR was 89.9% and 82.9% respectively compared to 76.5 and 68.0% for the ELISA. This study is the first to compare three different methods (ELISA, bg qPCR, nested PCR and sequencing at the gdh locus) to diagnose Jordanian patients suffering from giardiasis and to analyze their demographic data. Copyright © 2018 Elsevier Inc. All rights reserved.

  7. Correlation between real-time qPCR and development of strongyle eggs from cattle

    DEFF Research Database (Denmark)

    Drag, Markus; Nejsum, Peter; Höglund, Johan

    2014-01-01

    Differentiation of veterinary important parasitic strongyle eggs is time-consuming, because morphologically distinct third-stage larvae (L3) must be cultured for species/genus identification. A recently published qPCR technique provides a non-labour intensive method for detection and quantification...... of the two most important nematode eggs in cattle faeces. However, as quantification correlates with DNA content, quantification of copy numbers of the second internal transcribed spacer (ITS2) region is problematic as DNA content increases during egg development. The aim of this study was to assess...... the impact of oxygen availability and temperature on the multiplication of ITS2 copy numbers in O. ostertagi eggs. Fresh eggs were recovered from cattle faeces by sieving, flotation and entrapment in nylon-mesh filters and subsequently deposited in aliquots (n=18) of 5 ml distilled water with air circulation...

  8. Real-time pcr (qpcr) assay for rhizoctonia solani anastomoses group ag2-2 iiib

    International Nuclear Information System (INIS)

    Abbas, S.J.; Ahmad, B.

    2014-01-01

    Rhizoctonia solani anastomosis group AG2-2 IIIB is a severe sugar beet and maize pathogen. It causes crown and root rot disease which leads to yield losses world-wide. The soil-borne pathogen is difficult to detect and quantify by conventional methods. We developed a real-time PCR (qPCR) assay for the quantification of genomic DNA of Rhizoctonia solani AG2-2 IIIB based on the ITS region of rDNA genes. The limit of quantification of the assay is 1.8 pg genomic DNA. The amplification efficiency was 96.4. The assay will be helpful in the diagnoses of Rhizoctonia solani infection of sugar beet and maize roots and in the quantification of R. solani AG2-2 IIIB inoculum in plant debris and soil. (author)

  9. Sampling and Pooling Methods for Capturing Herd Level Antibiotic Resistance in Swine Feces using qPCR and CFU Approaches

    DEFF Research Database (Denmark)

    Schmidt, Gunilla Veslemøy; Mellerup, Anders; Christiansen, Lasse Engbo

    2015-01-01

    The aim of this article was to define the sampling level and method combination that captures antibiotic resistance at pig herd level utilizing qPCR antibiotic resistance gene quantification and culture-based quantification of antibiotic resistant coliform indicator bacteria. Fourteen qPCR assays...... for commonly detected antibiotic resistance genes were developed, and used to quantify antibiotic resistance genes in total DNA from swine fecal samples that were obtained using different sampling and pooling methods. In parallel, the number of antibiotic resistant coliform indicator bacteria was determined...... when comparing individual sampling and pooling methods. qPCR on pooled samples was found to be a good representative for the general resistance level in a pig herd compared to the coliform CFU counts. It had significantly reduced relative standard deviations compared to coliform CFU counts in the same...

  10. A ready-to-use duplex qPCR to detect Leishmania infantum DNA in naturally infected dogs.

    Science.gov (United States)

    Rampazzo, Rita de Cássia Pontello; Solcà, Manuela da Silva; Santos, Liliane Celestino Sales; Pereira, Lais de Novaes; Guedes, José Carlos Oliveira; Veras, Patrícia Sampaio Tavares; Fraga, Deborah Bittencourt Mothé; Krieger, Marco Aurélio; Costa, Alexandre Dias Tavares

    2017-11-15

    Canine visceral leishmaniasis (CVL) is a systemic disease caused by Leishmania infantum. A precise CVL diagnosis would allow for a faster and more specific treatment. Quantitative PCR (qPCR) is a sensitive and specific technique that can diagnose CVL and also monitor parasite load in the animal during the course of the infection or treatment. The aim of this study was to develop a ready-to-use (gelified and freezer-free) duplex qPCR for the identification of infected animals. We combined a new qPCR protocol that detects the canine 18S rRNA gene with an existing protocol for L. infantum kDNA detection, creating a duplex qPCR. This duplex method was then developed into a ready-to-use format. The performance of the duplex and singleplex reactions were compared in the traditional format (liquid and freezer-stored). Furthermore, the duplex qPCR performance was compared between the ready-to-use and traditional formats. The singleplex and new duplex qPCR exhibited the same detection limit in the traditional format (0.1 parasites/reaction). The ready-to-use format showed a detection limit of 1 parasite/reaction without affecting the reaction efficiency. The performance of the new qPCR protocol in the two formats was assessed using canine tissue samples from 82 dogs in an endemic CVL area that were previously characterized by standard serological and parasitological protocols. Splenic aspirates provided a higher rate of positivity (92.9%) followed by skin (50%) and blood (35.7%). The reported detection limits were observed for all tissues studied. Our results show that the amplification of L. infantum kDNA and canine DNA in a single tube, using either the traditional or ready-to-use format, exhibited the same diagnostic performance as amplification of the parasite kDNA alone. The detection of the host gene strengthens the qPCR results by confirming the presence and quality of DNA in the samples and the absence of polymerase inhibitors. The ready-to-use duplex qPCR format

  11. Monitoring bottlenose dolphin leukocyte cytokine mRNA responsiveness by qPCR

    Science.gov (United States)

    Eberle, Kirsten C.; Venn-Watson, Stephanie K.; Jensen, Eric D.; Porter, Tracy J.; Waters, Theresa E.; Sacco, Randy E.

    2017-01-01

    Both veterinarians caring for dolphins in managed populations and researchers monitoring wild populations use blood-based diagnostics to monitor bottlenose dolphin (Tursiops truncatus) health. Quantitative PCR (qPCR) can be used to assess cytokine transcription patterns of peripheral blood mononuclear cells (PBMC). This can supplement currently available blood tests with information on immune status. Full realization of this potential requires establishment of normal ranges of cytokine gene transcription levels in bottlenose dolphins. We surveyed four dolphins over the span of seven months by serial bleeds. PBMC were stimulated with phytohaemagglutinin (1, 5, and 10 μg/mL) and concanavalin A (1 μg/mL) for 48 H in vitro. RNA from these cultures was probed by qPCR using Tursiops truncatus-specific primers (IL-1α, IL-1β, IL-1RA, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p40, IL-13, IL-18, IFN-γ and TNF-α). Two blood samples from an additional bottlenose dolphin diagnosed with acute pulmonary disease add further perspective to the data. We observed that mitogen choice made a significant difference in the magnitude of gene transcription observed. On the other hand, most cytokines tested exhibited limited intra-animal variation. However, IL-6 and IL-12p40 differed between older and younger dolphins. Furthermore, the magnitude of mitogenic response clusters the tested cytokines into three groups. The data provide a reference for the selection of target cytokine mRNAs and their expected range of mitogen-stimulated cytokine gene transcription for future studies. PMID:29272269

  12. Molecular Quantification of Zooplankton Gut Content: The Case For qPCR

    Science.gov (United States)

    Frischer, M. E.; Walters, T. L.; Gibson, D. M.; Nejstgaard, J. C.; Troedsson, C.

    2016-02-01

    The ability to obtain information about feeding selectivity and rates in situ for zooplankton is vital for understanding the mechanisms structuring marine ecosystems. However, directly estimating feeding selection and rates of zooplankton, without bias, associated with culturing conditions has been notoriously difficult. A potential approach for addressing this problem is to target prey-specific DNA as a marker for prey ingestion and selection. In this study we report the development of a differential length amplification quantitative PCR (dla-qPCR) assay targeting the 18S rRNA gene to validate the use of a DNA-based approach to quantify consumption of specific plankton prey by the pelagic tunicate (doliolid) Dolioletta gegenbauri. Compared to copepods and other marine animals, the digestion of prey genomic DNA inside the gut of doliolids is low. This method minimizes potential underestimations, and therefore allows prey DNA to be used as an effective indicator of prey consumption. We also present an initial application of a qPCR-assay to estimate consumption of specific prey species on the southeastern continental shelf of the U.S., where doliolids stochastically bloom in response to upwelling events. Estimated feeding rates, based on qPCR, were in the same range as those estimated from clearance rates in laboratory feeding studies. In the field, consumption of specific prey, including the centric diatom Thalassiosira spp. was detected in the gut of wild caught D. gegenbauri at the levels consistent with their abundance in the water column at the time of collection. Thus, both experimental and field investigations support the hypothesis that a qPCR approach will be useful for the quantitative investigation of the in situ diet of D. gegenbauri without introduced bias' associated with cultivation.

  13. Monitoring bottlenose dolphin leukocyte cytokine mRNA responsiveness by qPCR.

    Directory of Open Access Journals (Sweden)

    Amelia Ruth Hofstetter

    Full Text Available Both veterinarians caring for dolphins in managed populations and researchers monitoring wild populations use blood-based diagnostics to monitor bottlenose dolphin (Tursiops truncatus health. Quantitative PCR (qPCR can be used to assess cytokine transcription patterns of peripheral blood mononuclear cells (PBMC. This can supplement currently available blood tests with information on immune status. Full realization of this potential requires establishment of normal ranges of cytokine gene transcription levels in bottlenose dolphins. We surveyed four dolphins over the span of seven months by serial bleeds. PBMC were stimulated with phytohaemagglutinin (1, 5, and 10 μg/mL and concanavalin A (1 μg/mL for 48 H in vitro. RNA from these cultures was probed by qPCR using Tursiops truncatus-specific primers (IL-1α, IL-1β, IL-1RA, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p40, IL-13, IL-18, IFN-γ and TNF-α. Two blood samples from an additional bottlenose dolphin diagnosed with acute pulmonary disease add further perspective to the data. We observed that mitogen choice made a significant difference in the magnitude of gene transcription observed. On the other hand, most cytokines tested exhibited limited intra-animal variation. However, IL-6 and IL-12p40 differed between older and younger dolphins. Furthermore, the magnitude of mitogenic response clusters the tested cytokines into three groups. The data provide a reference for the selection of target cytokine mRNAs and their expected range of mitogen-stimulated cytokine gene transcription for future studies.

  14. Pengaruh Green Marketing Hotel Terhadap Green Consumer Behavior

    OpenAIRE

    Yo Fernandez, Eunike Christe; Tjoanda, Evelyn

    2017-01-01

    Penelitian ini dilakukan untuk mengetahui pengaruh dari green marketing hotel terhadap green consumer behavior. Green marketing memiliki 3 dimensi, yaitu green product, green price, dan green promotion. Penelitian ini melibatkan 272 responden masyarakat Surabaya dan menggunakan metode regresi linear berganda. Hasil penelitian menunjukkan bahwa green product dan green price berpengaruh secara positif dan signifikan sedangkan green promotion berpengaruh namun tidak signifikan terhadap green con...

  15. A direct qPCR method for residual DNA quantification in monoclonal antibody drugs produced in CHO cells.

    Science.gov (United States)

    Hussain, Musaddeq

    2015-11-10

    Chinese hamster ovary (CHO) cells are the host cell of choice for manufacturing of monoclonal antibody (mAb) drugs in the biopharmaceutical industry. Host cell DNA is an impurity of such manufacturing process and must be controlled and monitored in order to ensure drug purity and safety. A conventional method for quantification of host residual DNA in drug requires extraction of DNA from the mAb drug substance with subsequent quantification of the extracted DNA using real-time PCR (qPCR). Here we report a method where the DNA extraction step is eliminated prior to qPCR. In this method, which we have named 'direct resDNA qPCR', the mAb drug substance is digested with a protease called KAPA in a 96-well PCR plate, the protease in the digest is then denatured at high temperature, qPCR reagents are added to the resultant reaction wells in the plate along with standards and controls in other wells of the same plate, and the plate subjected to qPCR for analysis of residual host DNA in the samples. This direct resDNA qPCR method for CHO is sensitive to 5.0fg of DNA with high precision and accuracy and has a wide linear range of determination. The method has been successfully tested with four mAbs drug, two IgG1 and two IgG4. Both the purified drug substance as well as a number of process intermediate samples, e.g., bioreactor harvest, Protein A column eluate and ion-exchange column eluates were tested. This method simplifies the residual DNA quantification protocol, reduces time of analysis and leads to increased assay sensitivity and development of automated high-throughput methods. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. AFLP fragment isolation technique as a method to produce random sequences for single nucleotide polymorphism discovery in the green turtle, Chelonia mydas.

    Science.gov (United States)

    Roden, Suzanne E; Dutton, Peter H; Morin, Phillip A

    2009-01-01

    The green sea turtle, Chelonia mydas, was used as a case study for single nucleotide polymorphism (SNP) discovery in a species that has little genetic sequence information available. As green turtles have a complex population structure, additional nuclear markers other than microsatellites could add to our understanding of their complex life history. Amplified fragment length polymorphism technique was used to generate sets of random fragments of genomic DNA, which were then electrophoretically separated with precast gels, stained with SYBR green, excised, and directly sequenced. It was possible to perform this method without the use of polyacrylamide gels, radioactive or fluorescent labeled primers, or hybridization methods, reducing the time, expense, and safety hazards of SNP discovery. Within 13 loci, 2547 base pairs were screened, resulting in the discovery of 35 SNPs. Using this method, it was possible to yield a sufficient number of loci to screen for SNP markers without the availability of prior sequence information.

  17. Green Computing

    Directory of Open Access Journals (Sweden)

    K. Shalini

    2013-01-01

    Full Text Available Green computing is all about using computers in a smarter and eco-friendly way. It is the environmentally responsible use of computers and related resources which includes the implementation of energy-efficient central processing units, servers and peripherals as well as reduced resource consumption and proper disposal of electronic waste .Computers certainly make up a large part of many people lives and traditionally are extremely damaging to the environment. Manufacturers of computer and its parts have been espousing the green cause to help protect environment from computers and electronic waste in any way.Research continues into key areas such as making the use of computers as energy-efficient as Possible, and designing algorithms and systems for efficiency-related computer technologies.

  18. Detection and quantification of Spirocerca lupi by HRM qPCR in fecal samples from dogs with spirocercosis.

    Science.gov (United States)

    Rojas, Alicia; Segev, Gilad; Markovics, Alex; Aroch, Itamar; Baneth, Gad

    2017-09-19

    Spirocerca lupi, the dog oesophageal nematode, causes a potentially fatal disease in domestic dogs, and is currently clinically diagnosed by coproscopy and oesophagoscopy. To date, a single molecular method, a semi-nested PCR, targeting the cox1 gene, has been developed to aid in the diagnosis of spirocercosis. The present study describes three novel high-resolution melt (HRM) quantitative PCR (qPCR) assays targeting fragments of the ITS1, 18S and cytb loci of S. lupi. The performance of these molecular assays in feces was compared to fecal flotation and to the previously described cox1 gene semi-nested PCR in 18 fecal samples from dogs with clinical oesophageal spirocercosis diagnosed by oesophagoscopy. The HRM qPCR for ITS1 and 18S were both able to detect 0.2 S. lupi eggs per gram (epg), while the HRM qPCR for the cytb and the semi-nested PCR for the cox1 detected 6 epg and 526 epg, respectively. Spirocerca lupi was detected in 61.1%, 44.4%, 27.8%, 11.1% and 5.6% of the fecal samples of dogs diagnosed with spirocercosis by using the ITS1 and 18S HRM qPCR assays, fecal flotation, cytb HRM qPCR and cox1 semi-nested PCR, respectively. All dogs positive by fecal flotation were also positive by ITS1 and 18S HRM qPCRs. Quantification of S. lupi eggs was successfully achieved in the HRM qPCRs and compared to the fecal flotation with no significant difference in the calculated concentrations between the HRM qPCRs that detected the 18S and ITS1 loci and the fecal flotation. The HRM qPCR for the 18S cross-amplified DNA from Toxocara canis and Toxascaris leonina. In contrast, the HRM qPCR for ITS1 did not cross-amplify DNA from other canine gastrointestinal parasites. This study presents two new molecular assays with significantly increased sensitivity for confirming and quantifying fecal S. lupi eggs. Of these, the HRM qPCR for ITS1 showed the best performance in terms of the limit of detection and absence of cross-amplification with other parasites. These assays will be

  19. Detection of Legionella by quantitative-polymerase chain reaction (qPCR) for monitoring and risk assessment

    DEFF Research Database (Denmark)

    Krøjgaard, Louise H.; Krogfelt, Karen A.; Albrechtsen, Hans-Jorgen

    2011-01-01

    Background: Culture and quantitative polymerase chain reaction (qPCR) assays for the detection of Legionella were compared on samples from a residential area before and after two interventions. A total of 84 samples were collected from shower hoses and taps as first flush samples and at constant...... temperature. Samples were grouped according to the origin of the sample, a) circulation water b) water from empty apartments c) water from shower hoses. The aims were to investigate the usefulness of qPCR compared to culture for monitoring remedial actions for elimination of Legionella bacteria and as a tool...

  20. Direct comparison of flow-FISH and qPCR as diagnostic tests for telomere length measurement in humans.

    Directory of Open Access Journals (Sweden)

    Fernanda Gutierrez-Rodrigues

    Full Text Available Telomere length measurement is an essential test for the diagnosis of telomeropathies, which are caused by excessive telomere erosion. Commonly used methods are terminal restriction fragment (TRF analysis by Southern blot, fluorescence in situ hybridization coupled with flow cytometry (flow-FISH, and quantitative PCR (qPCR. Although these methods have been used in the clinic, they have not been comprehensively compared. Here, we directly compared the performance of flow-FISH and qPCR to measure leukocytes' telomere length of healthy individuals and patients evaluated for telomeropathies, using TRF as standard. TRF and flow-FISH showed good agreement and correlation in the analysis of healthy subjects (R(2 = 0.60; p<0.0001 and patients (R(2 = 0.51; p<0.0001. In contrast, the comparison between TRF and qPCR yielded modest correlation for the analysis of samples of healthy individuals (R(2 = 0.35; p<0.0001 and low correlation for patients (R(2 = 0.20; p = 0.001; Bland-Altman analysis showed poor agreement between the two methods for both patients and controls. Quantitative PCR and flow-FISH modestly correlated in the analysis of healthy individuals (R(2 = 0.33; p<0.0001 and did not correlate in the comparison of patients' samples (R(2 = 0.1, p = 0.08. Intra-assay coefficient of variation (CV was similar for flow-FISH (10.8 ± 7.1% and qPCR (9.5 ± 7.4%; p = 0.35, but the inter-assay CV was lower for flow-FISH (9.6 ± 7.6% vs. 16 ± 19.5%; p = 0.02. Bland-Altman analysis indicated that flow-FISH was more precise and reproducible than qPCR. Flow-FISH and qPCR were sensitive (both 100% and specific (93% and 89%, respectively to distinguish very short telomeres. However, qPCR sensitivity (40% and specificity (63% to detect telomeres below the tenth percentile were lower compared to flow-FISH (80% sensitivity and 85% specificity. In the clinical setting, flow-FISH was more accurate, reproducible, sensitive, and specific in the measurement of human

  1. Green toxicology.

    Science.gov (United States)

    Maertens, Alexandra; Anastas, Nicholas; Spencer, Pamela J; Stephens, Martin; Goldberg, Alan; Hartung, Thomas

    2014-01-01

    Historically, early identification and characterization of adverse effects of industrial chemicals was difficult because conventional toxicological test methods did not meet R&D needs for rapid, relatively inexpensive methods amenable to small amounts of test material. The pharmaceutical industry now front-loads toxicity testing, using in silico, in vitro, and less demanding animal tests at earlier stages of product development to identify and anticipate undesirable toxicological effects and optimize product development. The Green Chemistry movement embraces similar ideas for development of less toxic products, safer processes, and less waste and exposure. Further, the concept of benign design suggests ways to consider possible toxicities before the actual synthesis and to apply some structure/activity rules (SAR) and in silico methods. This requires not only scientific development but also a change in corporate culture in which synthetic chemists work with toxicologists. An emerging discipline called Green Toxicology (Anastas, 2012) provides a framework for integrating the principles of toxicology into the enterprise of designing safer chemicals, thereby minimizing potential toxicity as early in production as possible. Green Toxicology`s novel utility lies in driving innovation by moving safety considerations to the earliest stage in a chemical`s lifecycle, i.e., to molecular design. In principle, this field is no different than other subdisciplines of toxicology that endeavor to focus on a specific area - for example, clinical, environmental or forensic toxicology. We use the same principles and tools to evaluate an existing substance or to design a new one. The unique emphasis is in using 21st century toxicology tools as a preventative strategy to "design out" undesired human health and environmental effects, thereby increasing the likelihood of launching a successful, sustainable product. Starting with the formation of a steering group and a series of workshops

  2. Green Gold

    International Nuclear Information System (INIS)

    Salamandra Martinez, Carlos

    2004-01-01

    The main purpose of this work is to offer a general panoramic of the processes or experiences pilot that are carried out in the Project Green Gold, as strategy of environmental sustainability and organizational invigoration in Choco, especially in the 12 communities of the municipalities of Tado and Condoto. It is also sought to offer a minimum of information on the techniques of handmade production and to show the possibilities to carry out in a rational way the use and use of the natural resources. The Project Green Gold is carried out by the Corporation Green Gold (COV) and co-financed with resources of international and national character, the intervention of the financial resources it achievement mainly for the use of clean processes in the extraction stages and metals benefit. The project is centered primarily in the absence of use of products or toxic substances as the mercury, fair trade, organizational invigoration, execution of 11 approaches and certification of the metals Gold and Platinum. The COV, it has come executing the proposal from the year 2001 with the premise of contributing to the balance between the rational exploitation of the natural resources and the conservation of the environment in the Choco. In the project they are used technical handmade characteristic of the region framed inside the mining activity and production activities are diversified in the productive family units. Those producing with the support of entities of juridical character, specify the necessary game rules for the extraction and products commercialization

  3. Greens of the European Green Capitals

    Science.gov (United States)

    Cömertler, Seval

    2017-10-01

    Well established and maintained green areas have a key role on reaching the high quality of life and sustainability in urban environments. Therefore, green areas must be carefully accounted and evaluated in the urban planning affairs. In this context, the European Green Capitals, which attach a great importance to the green areas, have a great potential to act as a role model for both small and big cities in all around the world. These leading cities (chronologically, Stockholm, Hamburg, Vitoria-Gasteiz, Nantes, Copenhagen, Bristol, Ljubljana, Essen and Nijmegen) are inspiring for the other cities which seek to achieve more sustainable and environmentally friendly places through green areas. From this point of view, the aim of this paper was to investigate the green areas of the European Green Capitals. The paper covered whole European Green Capitals, and the application form of each Green Capital was used as a primary data source. Consequently, the paper put forwarded that the European Green Capitals have considerably large amount and high proportion of green areas. Further, these cities provide an excellent access to the public green areas. As a result of abundant provision and proper distribution, the almost all citizens in most of the Green Capitals live within a distance of 300 meters to a green area. For further researches, the paper suggested that these green capitals should be investigated in terms of their efforts, measures, goals and plans, policies and implications to administer, to protect, to enhance and to expand the green areas.

  4. An easy 'one tube' method to estimate viability of Cryptosporidium oocysts using real-time qPCR

    NARCIS (Netherlands)

    Paziewska-Harris, A.; Schoone, G.; Schallig, H. D. F. H.

    2016-01-01

    Viability estimation of the highly resistant oocysts of Cryptosporidium remains a key issue for the monitoring and control of this pathogen. We present here a simple 'one tube' quantitative PCR (qPCR) protocol for viability estimation using a DNA extraction protocol which preferentially solubilizes

  5. Cryptosporidium diagnosis by qPCR in cats at Rio de Janeiro state, Brazil

    Directory of Open Access Journals (Sweden)

    Lara Patrícia Santos Carrasco

    2016-11-01

    Full Text Available ABSTRACT. Carrasco L.P.S., Oliveira R.L.S., Moreira C.M.R., Santos C.R.G.R., Corgozinho K.B. & Souza H.J.M. [Cryptosporidium diagnosis by qPCR in cats at Rio de Janeiro state, Brazil.] Diagnóstico de Cryptosporidium spp. pela técnica de qPCR em gatos no estado do Rio de Janeiro, Brasil. Revista Brasileira de Medicina Veterinária, 38(Supl.:22-26, 2016. Programa de Pós-Graduação em MedicinaVeteriná- ria, Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, BR-465, Km 7, Seropédica, RJ 23890-000, Brasil. E-mail: carrasco.lara@gmail.com Cryptosporidium spp. is recognized as an important etiologic agent of diarrhea in many countries. The aim of this study was to detect the presence of DNA of the parasite Cryptosporidium spp. in feces of cats with history of chronic diarrhea attended in the Feline Medicine Sector of the Veterinary Hospital of the Federal Rural University of Rio de Janeiro, by the polymerase chain reaction technique in real time (RT-PCR. In this study, 100 animals were admitted, of any breed or sex and from 8 weeks of age. As inclusion criteria, patients had to have diarrhea history for more than three weeks, with little success of clinical response to previously established therapies. From the samples obtained by collecting via washing the animal colon and spontaneous defecation, methods of direct examination of the feces, centrifugal flotation technique and real-time PCR were carried out. Of all cats selected for this study, 10% showed infection by Cryptosporidium spp. Most positive animals were aged over one year (70% and only 30% had up to one year old. Cats were 50% purebred and 50% were domestic short hair cats. The clinical signs presented by these cats at the time of consultation were diarrhea (60% and prolapsed rectum (40%. Four animals had co-infections with other enteropathogens (40%, such as Giardia, Toxocara sp. or Tritrichomonas fetus alone or combined. We concluded that infection by

  6. Analytical and diagnostic performance of a qPCR assay for Ichthyophonus spp. compared to the tissue culture 'gold standard'.

    Science.gov (United States)

    Lowe, Vanessa C; Hershberger, Paul K; Friedman, Carolyn S

    2018-06-04

    Parasites of the genus Ichthyophonus infect many fish species and have a non-uniform distribution within host tissues. Due in part to this uneven distribution, the comparative sensitivity and accuracy of using molecular-based detection methods versus culture to estimate parasite prevalence is under debate. We evaluated the analytical and diagnostic performance of an existing qPCR assay in comparison to the 'gold standard' culture method using Pacific herring Clupea pallasii with known exposure history. We determined that the assay is suitable for use in this host, and diagnostic specificity was consistently high (>98%) in both heart and liver tissues. Diagnostic sensitivity could not be fully assessed due to low infection rates, but our results suggest that qPCR is not as sensitive as culture under all circumstances. Diagnostic sensitivity of qPCR relative to culture is likely affected by the amount of sample processed. The prevalence values estimated by the 2 methods were not significantly different when sample amounts were equal (heart tissue), but when the assayed sample amounts were unequal (liver tissue), the culture method detected a significantly higher prevalence of the parasite than qPCR. Further, culture of liver also detected significantly more Ichthyophonus infections than culture of heart, suggesting that the density and distribution of parasites in tissues also plays a role in assay sensitivity. This sensitivity issue would be most problematic for fish with light infections. Although qPCR does not detect the presence of a live organism, DNA-based pathogen detection methods provide the opportunity for alternate testing strategies when culture is not possible.

  7. Identification of valid reference genes for the normalization of RT qPCR gene expression data in human brain tissue

    Directory of Open Access Journals (Sweden)

    Ravid Rivka

    2008-05-01

    Full Text Available Abstract Background Studies of gene expression in post mortem human brain can contribute to understanding of the pathophysiology of neurodegenerative diseases, including Alzheimer's disease (AD, Parkinson's disease (PD and dementia with Lewy bodies (DLB. Quantitative real-time PCR (RT qPCR is often used to analyse gene expression. The validity of results obtained using RT qPCR is reliant on accurate data normalization. Reference genes are generally used to normalize RT qPCR data. Given that expression of some commonly used reference genes is altered in certain conditions, this study aimed to establish which reference genes were stably expressed in post mortem brain tissue from individuals with AD, PD or DLB. Results The present study investigated the expression stability of 8 candidate reference genes, (ubiquitin C [UBC], tyrosine-3-monooxygenase [YWHAZ], RNA polymerase II polypeptide [RP II], hydroxymethylbilane synthase [HMBS], TATA box binding protein [TBP], β-2-microglobulin [B2M], glyceraldehyde-3-phosphate dehydrogenase [GAPDH], and succinate dehydrogenase complex-subunit A, [SDHA] in cerebellum and medial temporal gyrus of 6 AD, 6 PD, 6 DLB subjects, along with 5 matched controls using RT qPCR (TaqMan® Gene Expression Assays. Gene expression stability was analysed using geNorm to rank the candidate genes in order of decreasing stability in each disease group. The optimal number of genes recommended for accurate data normalization in each disease state was determined by pairwise variation analysis. Conclusion This study identified validated sets of mRNAs which would be appropriate for the normalization of RT qPCR data when studying gene expression in brain tissue of AD, PD, DLB and control subjects.

  8. Detection of Salmonella in Shellfish Using SYBR Green™ I-Based Real-Time Multiplexed PCR Assay Targeting invA and spvB

    KAUST Repository

    Gangwar, Maulshree; Waters, Alicia M.; Bej, Gautam A.; Bej, Asim K.; Mojib, Nazia

    2012-01-01

    A SYBR Green™ I-based real-time multiplexed PCR assay was developed targeting invA and spvB for the detection of Salmonella strains in shellfish after both hns and invA genes were identified in all Salmonella strains. Simultaneously, the 16S rRNA gene was used as a PCR internal amplification control (IAC). All 89 Salmonella strains tested in this study exhibited amplification of invA, whereas only 21 (23. 6 %) were PCR positive for spvB. The sensitivity of detection of all three targeted genes was 1 ng, which is equivalent to approximately 105 colony-forming unit (CFU) of Salmonella enterica. The analysis showed specific PCR products that were identified by reproducible melt temperature profiles (invA, 84. 27 ± 1. 7 °C; spvB, 88. 76 ± 1. 0 °C; and 16S rRNA gene, 87. 16 ± 0. 8 °C). The sensitivity of detection was 10 pg purified DNA (invA) or 105 CFU in 1 mL pure culture of S. enterica ATCC 14028. The above molecular detection method for Salmonella strains was successfully applied to the oyster homogenates (food matrix). An initial inoculum of 106 and 102 CFU Salmonella in 1 ml seeded oyster tissue homogenate was detected by multiplexed PCR for all three genes after 5 and 24 h of enrichment, respectively. Natural oysters isolated from Gulf of Mexico during the winter months exhibited negative PCR amplification results suggesting the absence of Salmonella. In contrast to conventional PCR, real-time multiplex PCR assay developed in this study is rapid and sensitive and will help Interstate Shellfish Sanitation Conference undertake appropriate measures to monitor Salmonella in oysters, thereby preventing disease outbreaks and consequently protecting consumer health. © 2012 Springer Science+Business Media, LLC.

  9. Detection of Salmonella in Shellfish Using SYBR Green™ I-Based Real-Time Multiplexed PCR Assay Targeting invA and spvB

    KAUST Repository

    Gangwar, Maulshree

    2012-09-23

    A SYBR Green™ I-based real-time multiplexed PCR assay was developed targeting invA and spvB for the detection of Salmonella strains in shellfish after both hns and invA genes were identified in all Salmonella strains. Simultaneously, the 16S rRNA gene was used as a PCR internal amplification control (IAC). All 89 Salmonella strains tested in this study exhibited amplification of invA, whereas only 21 (23. 6 %) were PCR positive for spvB. The sensitivity of detection of all three targeted genes was 1 ng, which is equivalent to approximately 105 colony-forming unit (CFU) of Salmonella enterica. The analysis showed specific PCR products that were identified by reproducible melt temperature profiles (invA, 84. 27 ± 1. 7 °C; spvB, 88. 76 ± 1. 0 °C; and 16S rRNA gene, 87. 16 ± 0. 8 °C). The sensitivity of detection was 10 pg purified DNA (invA) or 105 CFU in 1 mL pure culture of S. enterica ATCC 14028. The above molecular detection method for Salmonella strains was successfully applied to the oyster homogenates (food matrix). An initial inoculum of 106 and 102 CFU Salmonella in 1 ml seeded oyster tissue homogenate was detected by multiplexed PCR for all three genes after 5 and 24 h of enrichment, respectively. Natural oysters isolated from Gulf of Mexico during the winter months exhibited negative PCR amplification results suggesting the absence of Salmonella. In contrast to conventional PCR, real-time multiplex PCR assay developed in this study is rapid and sensitive and will help Interstate Shellfish Sanitation Conference undertake appropriate measures to monitor Salmonella in oysters, thereby preventing disease outbreaks and consequently protecting consumer health. © 2012 Springer Science+Business Media, LLC.

  10. Green Manufacturing

    Energy Technology Data Exchange (ETDEWEB)

    Patten, John

    2013-12-31

    Green Manufacturing Initiative (GMI): The initiative provides a conduit between the university and industry to facilitate cooperative research programs of mutual interest to support green (sustainable) goals and efforts. In addition to the operational savings that greener practices can bring, emerging market demands and governmental regulations are making the move to sustainable manufacturing a necessity for success. The funding supports collaborative activities among universities such as the University of Michigan, Michigan State University and Purdue University and among 40 companies to enhance economic and workforce development and provide the potential of technology transfer. WMU participants in the GMI activities included 20 faculty, over 25 students and many staff from across the College of Engineering and Applied Sciences; the College of Arts and Sciences' departments of Chemistry, Physics, Biology and Geology; the College of Business; the Environmental Research Institute; and the Environmental Studies Program. Many outside organizations also contribute to the GMI's success, including Southwest Michigan First; The Right Place of Grand Rapids, MI; Michigan Department of Environmental Quality; the Michigan Department of Energy, Labor and Economic Growth; and the Michigan Manufacturers Technical Center.

  11. Green chemistry

    International Nuclear Information System (INIS)

    Warner, John C.; Cannon, Amy S.; Dye, Kevin M.

    2004-01-01

    A grand challenge facing government, industry, and academia in the relationship of our technological society to the environment is reinventing the use of materials. To address this challenge, collaboration from an interdisciplinary group of stakeholders will be necessary. Traditionally, the approach to risk management of materials and chemicals has been through inerventions intended to reduce exposure to materials that are hazardous to health and the environment. In 1990, the Pollution Prevention Act encouraged a new tact-elimination of hazards at the source. An emerging approach to this grand challenge seeks to embed the diverse set of environmental perspectives and interests in the everyday practice of the people most responsible for using and creating new materials--chemists. The approach, which has come to be known as Green Chemistry, intends to eliminate intrinsic hazard itself, rather than focusing on reducing risk by minimizing exposure. This chapter addresses the representation of downstream environmental stakeholder interests in the upstream everyday practice that is reinventing chemistry and its material inputs, products, and waste as described in the '12 Principles of Green Chemistry'

  12. Green urbanity

    Directory of Open Access Journals (Sweden)

    Alenka Fikfak

    2012-01-01

    Full Text Available Tourism and other culture-based types of small business, which are the leitmotif in the planning of the Europark Ruardi, are becoming the guiding motif in the spatial development of urban centres that are influenced by dynamic transformation processes. The system should build upon the exploitation of both local and regional environmental features. This would encourage the quest for special environmental features, with an emphasis on their conservation, i.e. sustainable development, and connections in a wider context.The Europark is seen as a new strategic point of the Zasavje Region (the region of the central Sava Valley, which is linked to other important points in a region relevant for tourism. Due to the "smallness" of the region and/or the proximity of such points, development can be fast and effective. The interaction of different activities in space yields endless opportunities for users, who choose their own goals and priorities in the use of space. Four theme areas of the Europark area planning are envisaged. The organisation of activities is based on the composition of the mosaic field patterns, where green fields intertwine with areas of different, existing and new, urban functions. The fields of urban and recreation programmes are connected with a network of green areas and walking trails, along which theme park settings are arranged.

  13. Green shipping management

    CERN Document Server

    Lun, Y H Venus; Wong, Christina W Y; Cheng, T C E

    2016-01-01

    This book presents theory-driven discussion on the link between implementing green shipping practices (GSP) and shipping firm performance. It examines the shipping industry’s challenge of supporting economic growth while enhancing environmental performance. Consisting of nine chapters, the book covers topics such as the conceptualization of green shipping practices (GSPs), measurement scales for evaluating GSP implementation, greening capability, greening and performance relativity (GPR), green management practice, green shipping network, greening capacity, and greening propensity. In view of the increasing quest for environment protection in the shipping sector, this book provides a good reference for firms to understand and evaluate their capability in carrying out green operations on their shipping activities.

  14. From green architecture to architectural green

    DEFF Research Database (Denmark)

    Earon, Ofri

    2011-01-01

    that describes the architectural exclusivity of this particular architecture genre. The adjective green expresses architectural qualities differentiating green architecture from none-green architecture. Currently, adding trees and vegetation to the building’s facade is the main architectural characteristics...... they have overshadowed the architectural potential of green architecture. The paper questions how a green space should perform, look like and function. Two examples are chosen to demonstrate thorough integrations between green and space. The examples are public buildings categorized as pavilions. One......The paper investigates the topic of green architecture from an architectural point of view and not an energy point of view. The purpose of the paper is to establish a debate about the architectural language and spatial characteristics of green architecture. In this light, green becomes an adjective...

  15. Detection of Replication Competent Lentivirus Using a qPCR Assay for VSV-G

    Directory of Open Access Journals (Sweden)

    Lindsey M. Skrdlant

    2018-03-01

    Full Text Available Lentiviral vectors are a common tool used to introduce new and corrected genes into cell therapy products for treatment of human diseases. Although lentiviral vectors are ideal for delivery and stable integration of genes of interest into the host cell genome, they potentially pose risks to human health, such as integration-mediated transformation and generation of a replication competent lentivirus (RCL capable of infecting non-target cells. In consideration of the latter risk, all cell-based products modified by lentiviral vectors and intended for patient use must be tested for RCL prior to treatment of the patient. Current Food and Drug Administration (FDA guidelines recommend use of cell-based assays to this end, which can take up to 6 weeks for results. However, qPCR-based assays are a quick alternative for rapid assessment of RCL in products intended for fresh infusion. We describe here the development and qualification of a qPCR assay based on detection of envelope gene sequences (vesicular stomatitis virus G glycoprotein [VSV-G] for RCL in accordance with Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE guidelines. Our results demonstrate the sensitivity, linearity, specificity, and reproducibility of detection of VSV-G sequences, with a low false-positive rate. These procedures are currently being used in our phase 1 clinical investigations.

  16. A multimarker qPCR platform for the characterisation of endometrial cancer.

    Science.gov (United States)

    Supernat, Anna; Łapińska-Szumczyk, Sylwia; Majewska, Hanna; Gulczyński, Jacek; Biernat, Wojciech; Wydra, Dariusz; Zaczek, Anna J

    2014-02-01

    The molecular background of endometrial cancer (EC) has not been fully elucidated. In the present study, we developed a quantitative PCR (qPCR) platform to examine the gene dosages of the potential molecular markers MGB1, TOP2A, ERBB1-4, MYC, CCND1, ESR1 and PI3K. The platform was applied in samples collected from 157 EC patients (stage I-IV) to verify its clinical utility and to examine the diagnostic and prognostic significance of the analysed biomarkers. The gene dosage pattern of the ERBB family and its downstream effectors PI3K and MYC showed particularly strong correlations with clinicopathological data. The ERBB PI3K/Akt pathway was upregulated in 31 (20%) of 156 cases. Activation of the ERBB PI3K/Akt pathway was positively correlated with a higher stage (p=0.001), higher grade (p=0.001), histological type II disease (p=0.0003) and metastases (p=0.02). The implemented hierarchical clustering revealed that cluster 2 was characterised by high copy numbers of the studied genes. Cluster 2 was associated with shorter overall survival (p=0.05). The platform was found to be a fast and simple method for direct analysis of the genes involved in uterine carcinogenesis, making it feasible for EC biology characterisation.

  17. Simultaneous Detection of Ricin and Abrin DNA by Real-Time PCR (qPCR

    Directory of Open Access Journals (Sweden)

    Roman Wölfel

    2012-08-01

    Full Text Available Ricin and abrin are two of the most potent plant toxins known and may be easily obtained in high yield from the seeds using rather simple technology. As a result, both toxins are potent and available toxins for criminal or terrorist acts. However, as the production of highly purified ricin or abrin requires sophisticated equipment and knowledge, it may be more likely that crude extracts would be used by non-governmental perpetrators. Remaining plant-specific nucleic acids in these extracts allow the application of a real-time PCR (qPCR assay for the detection and identification of abrin or ricin genomic material. Therefore, we have developed a duplex real-time PCR assays for simultaneous detection of ricin and abrin DNA based on the OmniMix HS bead PCR reagent mixture. Novel primers and hybridization probes were designed for detection on a SmartCycler instrument by using 5′-nuclease technology. The assay was thoroughly optimized and validated in terms of analytical sensitivity. Evaluation of the assay sensitivity by probit analysis demonstrated a 95% probability of detection at 3 genomes per reaction for ricin DNA and 1.2 genomes per reaction for abrin DNA. The suitability of the assays was exemplified by detection of ricin and abrin contaminations in a food matrix.

  18. Empirical evaluation of humpback whale telomere length estimates; quality control and factors causing variability in the singleplex and multiplex qPCR methods

    DEFF Research Database (Denmark)

    Olsen, Morten Tange; Bérubé, Martine; Robbins, Jooke

    2012-01-01

    BACKGROUND:Telomeres, the protective cap of chromosomes, have emerged as powerful markers of biological age and life history in model and non-model species. The qPCR method for telomere length estimation is one of the most common methods for telomere length estimation, but has received recent...... steps of qPCR. In order to evaluate the utility of the qPCR method for telomere length estimation in non-model species, we carried out four different qPCR assays directed at humpback whale telomeres, and subsequently performed a rigorous quality control to evaluate the performance of each assay. RESULTS...... to 40% depending on assay and quantification method, however this variation only affected telomere length estimates in the worst performing assays. CONCLUSION:Our results suggest that seemingly well performing qPCR assays may contain biases that will only be detected by extensive quality control...

  19. Green business will remain green

    International Nuclear Information System (INIS)

    Marcan, P.

    2008-01-01

    It all started with two words. Climate change. The carbon dioxide trading scheme, which was the politicians' idea on solving the number one global problem, followed. Four years ago, when the project was begun, there was no data for project initiation. Quotas for polluters mainly from energy production and other energy demanding industries were distributed based on spreadsheets, maximum output and expected future development of economies. Slovak companies have had a chance to profit from these arrangements since 2005. Many of them took advantage of the situation and turned the excessive quotas into an extraordinary profit which often reached hundreds of million Sk. The fact that the price of free quotas offered for sale dropped basically to 0 in 2006 only proved that the initial distribution was too generous. And the market reacted to the first official measurements of emissions. Slovak companies also contributed to this development. However, when planning the maximum emission volumes for 2008-2012 period, in spite of the fact that actual data were available, their expectations were not realistic. A glance at the figures in the proposal of the Ministry of Environment is sufficient to realize that there will be no major change in the future. And so for many Slovak companies business with a green future will remain green for the next five years. The state decided to give to selected companies even more free space as far as emissions are concerned. The most privileged companies can expect quotas increased by tens of percent. (author)

  20. Green Power Partner Resources

    Science.gov (United States)

    EPA Green Power Partners can access tools and resources to help promote their green power commitments. Partners use these tools to communicate the benefits of their green power use to their customers, stakeholders, and the general public.

  1. Green Vehicle Guide

    Science.gov (United States)

    ... label Buy green. Save green. Learn about MPG math Discover fuel-saving tips Promote green ... U.S. consumers who have already purchased new vehicles under the fuel economy & greenhouse gas standard! More about the standards » Check ...

  2. Green Transformational Leadership and Green Performance: The Mediation Effects of Green Mindfulness and Green Self-Efficacy

    Directory of Open Access Journals (Sweden)

    Yu-Shan Chen

    2014-09-01

    Full Text Available No prior literature explores the influence of green transformational leadership on green performance, thus, this study develops a novel research framework to fill the research gap. This study investigates the influence of green transformational leadership on green performance and discusses the mediation effects of green mindfulness and green self-efficacy by means of structural equation modeling (SEM. The results indicate that green transformational leadership positively influences green mindfulness, green self-efficacy, and green performance. Moreover, this study demonstrates that the positive relationship between green transformational leadership and green performance is partially mediated by the two mediators: green mindfulness and green self-efficacy. It means that green transformational leadership can not only directly affect green performance positively but also indirectly affect it positively through green mindfulness and green self-efficacy. Therefore, firms need to raise their green transformational leadership, green mindfulness, and green self-efficacy to increase their green performance.

  3. Counting mycobacteria in infected human cells and mouse tissue: a comparison between qPCR and CFU.

    Directory of Open Access Journals (Sweden)

    Sharad Pathak

    Full Text Available Due to the slow growth rate and pathogenicity of mycobacteria, enumeration by traditional reference methods like colony counting is notoriously time-consuming, inconvenient and biohazardous. Thus, novel methods that rapidly and reliably quantify mycobacteria are warranted in experimental models to facilitate basic research, development of vaccines and anti-mycobacterial drugs. In this study we have developed quantitative polymerase chain reaction (qPCR assays for simultaneous quantification of mycobacterial and host DNA in infected human macrophage cultures and in mouse tissues. The qPCR method cannot discriminate live from dead bacteria and found a 10- to 100-fold excess of mycobacterial genomes, relative to colony formation. However, good linear correlations were observed between viable colony counts and qPCR results from infected macrophage cultures (Pearson correlation coefficient [r] for M. tuberculosis = 0.82; M. a. avium = 0.95; M. a. paratuberculosis = 0.91. Regression models that predict colony counts from qPCR data in infected macrophages were validated empirically and showed a high degree of agreement with observed counts. Similar correlation results were also obtained in liver and spleen homogenates of M. a. avium infected mice, although the correlations were distinct for the early phase (< day 9 post-infection and later phase (≥ day 20 post-infection liver r = 0.94 and r = 0.91; spleen r = 0.91 and r = 0.87, respectively. Interestingly, in the mouse model the number of live bacteria as determined by colony counts constituted a much higher proportion of the total genomic qPCR count in the early phase (geometric mean ratio of 0.37 and 0.34 in spleen and liver, respectively, as compared to later phase of infection (geometric mean ratio of 0.01 in both spleen and liver. Overall, qPCR methods offer advantages in biosafety, time-saving, assay range and reproducibility compared to colony counting. Additionally, the duplex format allows

  4. Central Region Green Infrastructure

    Data.gov (United States)

    Minnesota Department of Natural Resources — This Green Infrastructure data is comprised of 3 similar ecological corridor data layers ? Metro Conservation Corridors, green infrastructure analysis in counties...

  5. Advancing Our Understanding of the Etiologies and Mutational Landscapes of Basal Like, Luminal A, and Luminal B Breast Cancers

    Science.gov (United States)

    2016-10-01

    5-45, completed under Dr. Li’s supervision at FHCRC: a. Identification of controls, Months 5-45: General population controls with no prior history ...sequencing and analysis will be validated by SYBR Green based QPCR using Page 11 primers designed using Primer 3 to encompass the fusion junction on a...pools of multiplexed PCR products resequenced on Illumina next-generation sequencing equipment, we will screen any recurrently mutated gene appearing

  6. The green building envelope : Vertical greening

    NARCIS (Netherlands)

    Ottelé, M.

    2011-01-01

    Planting on roofs and façades is one of the most innovative and fastest developing fields of green technologies with respect to the built environment and horticulture. This thesis is focused on vertical greening of structures and to the multi-scale benefits of vegetation. Vertical green can improve

  7. MeltMan: Optimization, Evaluation, and Universal Application of a qPCR System Integrating the TaqMan qPCR and Melting Analysis into a Single Assay

    Science.gov (United States)

    Nagy, Alexander; Černíková, Lenka; Vitásková, Eliška; Křivda, Vlastimil; Dán, Ádám; Dirbáková, Zuzana; Jiřincová, Helena; Procházka, Bohumír; Sedlák, Kamil; Havlíčková, Martina

    2016-01-01

    In the present work, we optimised and evaluated a qPCR system integrating 6-FAM (6-carboxyfluorescein)-labelled TaqMan probes and melting analysis using the SYTO 82 (S82) DNA binding dye in a single reaction. We investigated the influence of the S82 on various TaqMan and melting analysis parameters and defined its optimal concentration. In the next step, the method was evaluated in 36 different TaqMan assays with a total of 729 paired reactions using various DNA and RNA templates, including field specimens. In addition, the melting profiles of interest were correlated with the electrophoretic patterns. We proved that the S82 is fully compatible with the FAM-TaqMan system. Further, the advantages of this approach in routine diagnostic TaqMan qPCR were illustrated with practical examples. These included solving problems with flat or other atypical amplification curves or even false negativity as a result of probe binding failure. Our data clearly show that the integration of the TaqMan qPCR and melting analysis into a single assay provides an additional control option as well as the opportunity to perform more complex analyses, get more data from the reactions, and obtain analysis results with higher confidence. PMID:27031831

  8. Reference gene validation for qPCR in rat carotid body during postnatal development

    Directory of Open Access Journals (Sweden)

    Carroll John L

    2011-10-01

    Full Text Available Abstract Background The carotid bodies are the main arterial oxygen chemoreceptors in mammals. Afferent neural output from the carotid bodies to brainstem respiratory and cardiovascular nuclei provides tonic input and mediates important protective responses to acute and chronic hypoxia. It is widely accepted that the selection of reference genes for mRNA normalization in quantitative real-time PCR must be validated for a given tissue and set of conditions. This is particularly important for studies in carotid body during early postnatal maturation as the arterial oxygen tension undergoes major changes from fetal to postnatal life, which may affect reference gene expression. In order to determine the most stable and suitable reference genes for the study of rat carotid body during development, six commonly used reference genes, β-actin, RPII (RNA polymerase II, PPIA (peptidyl-proyl-isomerase A, TBP (TATA-box binding protein, GAPDH, and 18s rRNA, were evaluated in two age groups (P0-1 and P14-16 under three environmental oxygen conditions (normoxia, chronic hypoxia and chronic hyperoxia using the three most commonly used software programs, geNorm, NormFinder and BestKeeper. Findings The three programs produced similar results but the reference gene rankings were not identical between programs or experimental conditions. Overall, 18s rRNA was the least stable reference gene for carotid body and, when hyperoxia and/or hypoxia conditions were included, actin was similarly unstable. Conclusions Reference or housekeeping gene expression for qPCR studies of carotid body during postnatal development may vary with developmental stage and environmental conditions. Selection of the best reference gene or combination of reference genes for carotid body development studies should take environmental conditions into account. Two commonly used reference genes, 18s rRNA and actin, may be unsuitable for studies of carotid body maturation, especially if the study

  9. Elasmobranch qPCR reference genes: a case study of hypoxia preconditioned epaulette sharks

    Directory of Open Access Journals (Sweden)

    Ashton Kevin J

    2010-04-01

    Full Text Available Abstract Background Elasmobranch fishes are an ancient group of vertebrates which have high potential as model species for research into evolutionary physiology and genomics. However, no comparative studies have established suitable reference genes for quantitative PCR (qPCR in elasmobranchs for any physiological conditions. Oxygen availability has been a major force shaping the physiological evolution of vertebrates, especially fishes. Here we examined the suitability of 9 reference candidates from various functional categories after a single hypoxic insult or after hypoxia preconditioning in epaulette shark (Hemiscyllium ocellatum. Results Epaulette sharks were caught and exposed to hypoxia. Tissues were collected from 10 controls, 10 individuals with single hypoxic insult and 10 individuals with hypoxia preconditioning (8 hypoxic insults, 12 hours apart. We produced sequence information for reference gene candidates and monitored mRNA expression levels in four tissues: cerebellum, heart, gill and eye. The stability of the genes was examined with analysis of variance, geNorm and NormFinder. The best ranking genes in our study were eukaryotic translation elongation factor 1 beta (eef1b, ubiquitin (ubq and polymerase (RNA II (DNA directed polypeptide F (polr2f. The performance of the ribosomal protein L6 (rpl6 was tissue-dependent. Notably, in one tissue the analysis of variance indicated statistically significant differences between treatments for genes that were ranked as the most stable candidates by reference gene software. Conclusions Our results indicate that eef1b and ubq are generally the most suitable reference genes for the conditions and tissues in the present epaulette shark studies. These genes could also be potential reference gene candidates for other physiological studies examining stress in elasmobranchs. The results emphasise the importance of inter-group variation in reference gene evaluation.

  10. How Green is 'Green' Energy?

    Science.gov (United States)

    Gibson, Luke; Wilman, Elspeth N; Laurance, William F

    2017-12-01

    Renewable energy is an important piece of the puzzle in meeting growing energy demands and mitigating climate change, but the potentially adverse effects of such technologies are often overlooked. Given that climate and ecology are inextricably linked, assessing the effects of energy technologies requires one to consider their full suite of global environmental concerns. We review here the ecological impacts of three major types of renewable energy - hydro, solar, and wind energy - and highlight some strategies for mitigating their negative effects. All three types can have significant environmental consequences in certain contexts. Wind power has the fewest and most easily mitigated impacts; solar energy is comparably benign if designed and managed carefully. Hydropower clearly has the greatest risks, particularly in certain ecological and geographical settings. More research is needed to assess the environmental impacts of these 'green' energy technologies, given that all are rapidly expanding globally. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Practical utilization of recombinant AAV vector reference standards: focus on vector genomes titration by free ITR qPCR

    Directory of Open Access Journals (Sweden)

    Susan D'Costa

    2016-01-01

    Full Text Available Clinical trials using recombinant adeno-associated virus (rAAV vectors have demonstrated efficacy and a good safety profile. Although the field is advancing quickly, vector analytics and harmonization of dosage units are still a limitation for commercialization. AAV reference standard materials (RSMs can help ensure product safety by controlling the consistency of assays used to characterize rAAV stocks. The most widely utilized unit of vector dosing is based on the encapsidated vector genome. Quantitative polymerase chain reaction (qPCR is now the most common method to titer vector genomes (vg; however, significant inter- and intralaboratory variations have been documented using this technique. Here, RSMs and rAAV stocks were titered on the basis of an inverted terminal repeats (ITRs sequence-specific qPCR and we found an artificial increase in vg titers using a widely utilized approach. The PCR error was introduced by using single-cut linearized plasmid as the standard curve. This bias was eliminated using plasmid standards linearized just outside the ITR region on each end to facilitate the melting of the palindromic ITR sequences during PCR. This new “Free-ITR” qPCR delivers vg titers that are consistent with titers obtained with transgene-specific qPCR and could be used to normalize in-house product-specific AAV vector standards and controls to the rAAV RSMs. The free-ITR method, including well-characterized controls, will help to calibrate doses to compare preclinical and clinical data in the field.

  12. Practical utilization of recombinant AAV vector reference standards: focus on vector genomes titration by free ITR qPCR.

    Science.gov (United States)

    D'Costa, Susan; Blouin, Veronique; Broucque, Frederic; Penaud-Budloo, Magalie; François, Achille; Perez, Irene C; Le Bec, Christine; Moullier, Philippe; Snyder, Richard O; Ayuso, Eduard

    2016-01-01

    Clinical trials using recombinant adeno-associated virus (rAAV) vectors have demonstrated efficacy and a good safety profile. Although the field is advancing quickly, vector analytics and harmonization of dosage units are still a limitation for commercialization. AAV reference standard materials (RSMs) can help ensure product safety by controlling the consistency of assays used to characterize rAAV stocks. The most widely utilized unit of vector dosing is based on the encapsidated vector genome. Quantitative polymerase chain reaction (qPCR) is now the most common method to titer vector genomes (vg); however, significant inter- and intralaboratory variations have been documented using this technique. Here, RSMs and rAAV stocks were titered on the basis of an inverted terminal repeats (ITRs) sequence-specific qPCR and we found an artificial increase in vg titers using a widely utilized approach. The PCR error was introduced by using single-cut linearized plasmid as the standard curve. This bias was eliminated using plasmid standards linearized just outside the ITR region on each end to facilitate the melting of the palindromic ITR sequences during PCR. This new "Free-ITR" qPCR delivers vg titers that are consistent with titers obtained with transgene-specific qPCR and could be used to normalize in-house product-specific AAV vector standards and controls to the rAAV RSMs. The free-ITR method, including well-characterized controls, will help to calibrate doses to compare preclinical and clinical data in the field.

  13. Unfolding Green Defense

    DEFF Research Database (Denmark)

    Larsen, Kristian Knus

    2015-01-01

    In recent years, many states have developed and implemented green solutions for defense. Building on these initiatives NATO formulated the NATO Green Defence Framework in 2014. The framework provides a broad basis for cooperation within the Alliance on green solutions for defense. This report aims...... to inform and support the further development of green solutions by unfolding how green technologies and green strategies have been developed and used to handle current security challenges. The report, initially, focuses on the security challenges that are being linked to green defense, namely fuel...... consumption in military operations, defense expenditure, energy security, and global climate change. The report then proceeds to introduce the NATO Green Defence Framework before exploring specific current uses of green technologies and green strategies for defense. The report concludes that a number...

  14. Viability qPCR, a new tool for Legionella risk management.

    Science.gov (United States)

    Lizana, X; López, A; Benito, S; Agustí, G; Ríos, M; Piqué, N; Marqués, A M; Codony, F

    2017-11-01

    Viability quantitative Polymerase Chain Reaction (v-qPCR) is a recent analytical approach for only detecting live microorganisms by DNA amplification-based methods This approach is based on the use of a reagent that irreversibly fixes dead cells DNA. In this study, we evaluate the utility of v-qPCR versus culture method for Legionellosis risk management. The present study was performed using 116 real samples. Water samples were simultaneously analysed by culture, v-qPCR and qPCR methods. Results were compared by means of a non-parametric test. In 11.6% of samples using both methods (culture method and v-qPCR) results were positive, in 50.0% of samples both methods gave rise to negative results. As expected, equivalence between methods was not observed in all cases, as in 32.1% of samples positive results were obtained by v-qPCR and all of them gave rise to negative results by culture. Only in 6.3% of samples, with very low Legionella levels, was culture positive and v-qPCR negative. In 3.5% of samples, overgrowth of other bacteria did not allow performing the culture. When comparing both methods, significant differences between culture and v-qPCR were in the samples belonging to the cooling towers-evaporative condensers group. The v-qPCR method detected greater presence and obtained higher concentrations of Legionella spp. (p<0.001). Otherwise, no significant differences between methods were found in the rest of the groups. The v-qPCR method can be used as a quick tool to evaluate Legionellosis risk, especially in cooling towers-evaporative condensers, where this technique can detect higher levels than culture. The combined interpretation of PCR results along with the ratio of live cells is proposed as a tool for understanding the sample context and estimating the Legionellosis risk potential according to 4 levels of hierarchy. Copyright © 2017 Elsevier GmbH. All rights reserved.

  15. Selection of reference genes for qPCR in hairy root cultures of peanut

    Directory of Open Access Journals (Sweden)

    Medrano Giuliana

    2011-10-01

    Full Text Available Abstract Background Hairy root cultures produced via Agrobacterium rhizogenes-mediated transformation have emerged as practical biological models to elucidate the biosynthesis of specialized metabolites. To effectively understand the expression patterns of the genes involved in the metabolic pathways of these compounds, reference genes need to be systematically validated under specific experimental conditions as established by the MIQE (Minimum Information for Publication of Quantitative Real-Time PCR Experiments guidelines. In the present report we describe the first validation of reference genes for RT-qPCR in hairy root cultures of peanut which produce stilbenoids upon elicitor treatments. Results A total of 21 candidate reference genes were evaluated. Nineteen genes were selected based on previous qPCR studies in plants and two were from the T-DNAs transferred from A. rhizogenes. Nucleotide sequences of peanut candidate genes were obtained using their homologous sequences in Arabidopsis. To identify the suitable primers, calibration curves were obtained for each candidate reference gene. After data analysis, 12 candidate genes meeting standard efficiency criteria were selected. The expression stability of these genes was analyzed using geNorm and NormFinder algorithms and a ranking was established based on expression stability of the genes. Candidate reference gene expression was shown to have less variation in methyl jasmonate (MeJA treated root cultures than those treated with sodium acetate (NaOAc. Conclusions This work constitutes the first effort to validate reference genes for RT-qPCR in hairy roots. While these genes were selected under conditions of NaOAc and MeJA treatment, we anticipate these genes to provide good targets for reference genes for hairy roots under a variety of stress conditions. The lead reference genes were a gene encoding for a TATA box binding protein (TBP2 and a gene encoding a ribosomal protein (RPL8C. A

  16. Green industrial policy

    OpenAIRE

    Dani Rodrik

    2014-01-01

    Green growth requires green technologies: production techniques that economize on exhaustible resources and emit fewer greenhouse gases. The availability of green technologies both lowers social costs in the transition to a green growth path and helps achieve a satisfactory rate of material progress under that path. The theoretical case in favour of using industrial policy to facilitate green growth is quite strong. Economists’ traditional scepticism on industrial policy is grounded instead o...

  17. Apparent polyploidization after gamma irradiation: pitfalls in the use of quantitative polymerase chain reaction (qPCR) for the estimation of mitochondrial and nuclear DNA gene copy numbers.

    Science.gov (United States)

    Kam, Winnie W Y; Lake, Vanessa; Banos, Connie; Davies, Justin; Banati, Richard

    2013-05-30

    Quantitative polymerase chain reaction (qPCR) has been widely used to quantify changes in gene copy numbers after radiation exposure. Here, we show that gamma irradiation ranging from 10 to 100 Gy of cells and cell-free DNA samples significantly affects the measured qPCR yield, due to radiation-induced fragmentation of the DNA template and, therefore, introduces errors into the estimation of gene copy numbers. The radiation-induced DNA fragmentation and, thus, measured qPCR yield varies with temperature not only in living cells, but also in isolated DNA irradiated under cell-free conditions. In summary, the variability in measured qPCR yield from irradiated samples introduces a significant error into the estimation of both mitochondrial and nuclear gene copy numbers and may give spurious evidence for polyploidization.

  18. Multi-laboratory survey of qPCR enterococci analysis method performance in U.S. coastal and inland surface waters

    Data.gov (United States)

    U.S. Environmental Protection Agency — Quantitative polymerase chain reaction (qPCR) has become a frequently used technique for quantifying enterococci in recreational surface waters, but there are...

  19. Green corridors basics

    DEFF Research Database (Denmark)

    Panagakos, George

    2016-01-01

    SuperGreen project, which aimed at advancing the green corridor concept through a benchmarking exercise involving Key Performance Indicators (KPIs). The chapter discusses the available definitions of green corridors and identifies the characteristics that distinguish a green corridor from any other...... efficient surface transportation corridor. After providing examples of green corridor projects in Europe, it focuses on the KPIs that have been proposed by various projects for monitoring the performance of a freight corridor. Emphasis is given to the SuperGreen KPIs, covering the economic, technical...

  20. Limitations of Using Propidium Monoazide with qPCR to Discriminate between Live and Dead in Biofilm Samples

    Directory of Open Access Journals (Sweden)

    Michael J. Taylor

    2014-01-01

    Full Text Available Accurately quantifying Legionella for regulatory purposes to protect public health is essential. Real-time PCR (qPCR has been proposed as a better method for detecting and enumerating Legionella in samples than conventional culture method. However, since qPCR amplifies any target DNA in the sample, the technique's inability to discriminate between live and dead cells means that counts are generally significantly overestimated. Propidium monoazide (PMA has been used successfully in qPCR to aid live/dead discrimination. We tested PMA use as a method to count only live Legionella cells in samples collected from a modified chemostat that generates environmentally comparable samples. Counts from PMA-treated samples that were pretreated with either heat or three types of disinfectants (to kill the cells were highly variable, with the only consistent trend being the relationship between biofilm mass and numbers of Legionella cells. Two possibilities explain this result: 1. PMA treatment worked and the subsequent muted response of Legionella to disinfection treatment is a factor of biofilm/microbiological effects; although this does not account for the relationship between the amount of biofilm sampled and the viable Legionella count as determined by PMA-qPCR; or 2. PMA treatment did not work, and any measured decrease or increase in detectable Legionella is because of other factors affecting the method. This is the most likely explanation for our results, suggesting that higher concentrations of PMA might be needed to compensate for the presence of other compounds in an environmental sample or that lower amounts of biofilm need to be sampled. As PMA becomes increasingly toxic at higher concentrations and is very expensive, augmenting the method to include higher PMA concentrations is both counterproductive and cost prohibitive. Conversely, if smaller volumes of biofilm are used, the reproducibility of the method is reduced. Our results suggest that

  1. Development of a multiplex Q-PCR to detect Trichoderma harzianum Rifai strain T22 in plant roots.

    Science.gov (United States)

    Horn, Ivo R; van Rijn, Menno; Zwetsloot, Tom J J; Basmagi, Said; Dirks-Mulder, Anita; van Leeuwen, Willem B; Ravensberg, Willem J; Gravendeel, Barbara

    2016-02-01

    The fungal species Trichoderma harzianum is widely used as a biological agent in crop protection. To verify the continued presence of this fungus on plant roots manually inoculated with T. harzianum strain T22, a Q-PCR was designed using specific probes for this particular strain. To develop these molecular diagnostic tools, genome mining was first carried out to retrieve putative new regions by which different strains of T. harzianum could be distinguished. Subsequently, Sanger sequencing of the L-aminoacid oxidase gene (aox1) in T. harzianum was applied to determine the mutations differing between various strains isolated from the Trichoderma collection of Koppert Biological Systems. Based on the sequence information obtained, a set of hydrolysis probes was subsequently developed which discriminated T. harzianum T22 strains varying in only a single nucleotide. Probes designed for two strains uniquely recognized the respective strains in Q-PCR with a detection limit of 12,5ng DNA. Titration assays in which T. harzianum DNA from distinct strains was varied further underscored the specificity of the probes. Lastly, fungal DNA extracted from roots of greenhouse cultured tomato plants was analyzed using the probe-based assay. DNA from T. harzianum strain T22 could readily be identified on roots of greenhouse reared tomato plants inoculated with varying concentrations up to one week after treatment with a detection limit of 3e6 colony forming units of T. harzianum T22. We conclude that the Q-PCR method is a reliable and robust method for assessing the presence and quantity of T. harzianum strain T22 in manually inoculated plant material. Our method provides scope for the development of DNA based strain specific identification of additional strains of Trichoderma and other fungal biological control agents. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. A pipeline to determine RT-QPCR control genes for evolutionary studies: application to primate gene expression across multiple tissues.

    Directory of Open Access Journals (Sweden)

    Olivier Fedrigo

    Full Text Available Because many species-specific phenotypic differences are assumed to be caused by differential regulation of gene expression, many recent investigations have focused on measuring transcript abundance. Despite the availability of high-throughput platforms, quantitative real-time polymerase chain reaction (RT-QPCR is often the method of choice because of its low cost and wider dynamic range. However, the accuracy of this technique heavily relies on the use of multiple valid control genes for normalization. We created a pipeline for choosing genes potentially useful as RT-QPCR control genes for measuring expression between human and chimpanzee samples across multiple tissues, using published microarrays and a measure of tissue-specificity. We identified 13 genes from the pipeline and from commonly used control genes: ACTB, USP49, ARGHGEF2, GSK3A, TBP, SDHA, EIF2B2, GPDH, YWHAZ, HPTR1, RPL13A, HMBS, and EEF2. We then tested these candidate genes and validated their expression stability across species. We established the rank order of the most preferable set of genes for single and combined tissues. Our results suggest that for at least three tissues (cerebral cortex, liver, and skeletal muscle, EIF2B2, EEF2, HMBS, and SDHA are useful genes for normalizing human and chimpanzee expression using RT-QPCR. Interestingly, other commonly used control genes, including TBP, GAPDH, and, especially ACTB do not perform as well. This pipeline could be easily adapted to other species for which expression data exist, providing taxonomically appropriate control genes for comparisons of gene expression among species.

  3. Green Power Partnership 100 Green Power Users

    Science.gov (United States)

    EPA's Green Power Partnership is a voluntary program designed to reduce the environmental impact of electricity generation by promoting renewable energy. Partners on this list use green power to meet 100 of their U.S. organization-wide electricity use.

  4. Urban Greening Bay Area

    Science.gov (United States)

    Information about the San Francisco Bay Water Quality Project (SFBWQP) Urban Greening Bay Area, a large-scale effort to re-envision urban landscapes to include green infrastructure (GI) making communities more livable and reducing stormwater runoff.

  5. Tribal Green Building Toolkit

    Science.gov (United States)

    This Tribal Green Building Toolkit (Toolkit) is designed to help tribal officials, community members, planners, developers, and architects develop and adopt building codes to support green building practices. Anyone can use this toolkit!

  6. Green Power Partner List

    Science.gov (United States)

    The U.S. EPA's Green Power Partnership is a voluntary program designed to reduce the environmental impact of electricity generation by promoting renewable energy. There are thousands of Green Power Partners, all listed on this page.

  7. Green Power Communities

    Science.gov (United States)

    GPCs are towns, villages, cities, counties, or tribal governments in which the local government, businesses, and residents collectively use green power in amounts that meet or exceed EPA's Green Power Community purchase requirements.

  8. Blue-Green Algae

    Science.gov (United States)

    ... that taking a specific blue-green algae product (Super Blue-Green Algae, Cell Tech, Klamath Falls, OR) ... system. Premenstrual syndrome (PMS). Depression. Digestion. Heart disease. Memory. Wound healing. Other conditions. More evidence is needed ...

  9. Green Bank Observatory (GBO)

    Data.gov (United States)

    Federal Laboratory Consortium — The largest fully steerable telescope in the world - the Robert C. Byrd Green Bank Telescope (GBT), began observations in Green Bank, West Virginia in 2000and is a...

  10. Green Infrastructure Modeling Toolkit

    Science.gov (United States)

    Green infrastructure, such as rain gardens, green roofs, porous pavement, cisterns, and constructed wetlands, is becoming an increasingly attractive way to recharge aquifers and reduce the amount of stormwater runoff that flows into wastewater treatment plants or into waterbodies...

  11. Show Me the Green

    Science.gov (United States)

    Norbury, Keith

    2013-01-01

    Gone are the days when green campus initiatives were a balm to the soul and a drain on the wallet. Today's environmental initiatives are all about saving lots of green--in every sense of the word. The environmental benefits of green campus projects--whether wind turbines or better insulation--are pretty clear. Unfortunately, in today's…

  12. Green roof Malta

    OpenAIRE

    Gatt, Antoine

    2015-01-01

    In Malta, buildings cover one third of the Island, leaving greenery in the dirt track. Green roofs are one way to bring plants back to urban areas with loads of benefits. Antoine Gatt, who manages the LifeMedGreenRoof project at the University of Malta, tells us more. http://www.um.edu.mt/think/green-roof-malta/

  13. EPA's Green Roof Research

    Science.gov (United States)

    This is a presentation on the basics of green roof technology. The presentation highlights some of the recent ORD research projects on green roofs and provices insight for the end user as to the benefits for green roof technology. It provides links to currently available EPA re...

  14. In the Green

    Science.gov (United States)

    Kennedy, Mike

    2011-01-01

    Education officials used to debate whether they could afford to pursue green design and construction. Now the green movement has gained a foothold not just in education, but in society at large, and the prevailing attitude seems to have shifted. Can schools afford "not" to go green? As budgets are slashed repeatedly, education administrators must…

  15. The green agenda

    CERN Document Server

    Calder, Alan

    2009-01-01

    This business guide to Green IT was written to introduce, to a business audience, the opposing groups and the key climate change concepts, to provide an overview of a Green IT strategy and to set out a straightforward, bottom line-orientated Green IT action plan.

  16. The Green Man

    Science.gov (United States)

    Watson-Newlin, Karen

    2010-01-01

    The Jolly Green Giant. Robin Hood. The Bamberg Cathedral. Tales of King Arthur. Ecology. What do they have in common? What legends and ancient myths are shrouded in the tales of the Green Man? Most often perceived as an ancient Celtic symbol as the god of spring and summer, the Green Man disappears and returns year after year, century after…

  17. A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages

    DEFF Research Database (Denmark)

    Muhammed, Musemma Kedir; Krych, Lukasz; Nielsen, Dennis Sandris

    2017-01-01

    simultaneous quantitative detection of Lc. lactis 936 (now SK1virus), P335, c2 (now C2virus) and Leuconostoc phage groups. Component assays are designed to have high efficiencies and nearly the same dynamic detection ranges, i.e., from 1.1 x 105 to 1.1 x 101 phage genomes per reaction, which corresponds to 9 x......Simultaneous quantitative detection of Lactococcus (Lc.) lactis and Leuconostoc species bacteriophages (phages) has not been reported in dairies using undefined mixed-strain DL-starters, probably due to the lack of applicable methods. We optimized a high-throughput qPCR system that allows...... 107 to 9 x 103 phage particles mL-1 without any additional up-concentrating steps. The amplification efficiencies of the corresponding assays were 100.1±2.6, 98.7±2.3, 101.0±2.3 and 96.2±6.2. The qPCR system was tested on samples obtained from a dairy plant that employed traditional mother...

  18. Seasonal Dynamics of Microcystis spp. and Their Toxigenicity as Assessed by qPCR in a Temperate Reservoir

    Directory of Open Access Journals (Sweden)

    Agostinho Antunes

    2011-09-01

    Full Text Available Blooms of toxic cyanobacteria are becoming increasingly frequent, mainly due to water quality degradation. This work applied qPCR as a tool for early warning of microcystin(MC-producer cyanobacteria and risk assessment of water supplies. Specific marker genes for cyanobacteria, Microcystis and MC-producing Microcystis, were quantified to determine the genotypic composition of the natural Microcystis population. Correlations between limnological parameters, pH, water temperature, dissolved oxygen and conductivity and MC concentrations as well as Microcystis abundance were assessed. A negative significant correlation was observed between toxic (with mcy genes to non-toxic (without mcy genes genotypes ratio and the overall Microcystis density. The highest proportions of toxic Microcystis genotypes were found 4–6 weeks before and 8–10 weeks after the peak of the bloom, with the lowest being observed at its peak. These results suggest positive selection of non-toxic genotypes under favorable environmental growth conditions. Significant positive correlations could be found between quantity of toxic genotypes and MC concentration, suggesting that the method applied can be useful to predict potential MC toxicity risk. No significant correlation was found between the limnological parameters measured and MC concentrations or toxic genotypes proportions indicating that other abiotic and biotic factors should be governing MC production and toxic genotypes dynamics. The qPCR method here applied is useful to rapidly estimate the potential toxicity of environmental samples and so, it may contribute to the more efficient management of water use in eutrophic systems.

  19. SNPase-ARMS qPCR: Ultrasensitive Mutation-Based Detection of Cell-Free Tumor DNA in Melanoma Patients.

    Directory of Open Access Journals (Sweden)

    Julia Stadler

    Full Text Available Cell-free circulating tumor DNA in the plasma of cancer patients has become a common point of interest as indicator of therapy options and treatment response in clinical cancer research. Especially patient- and tumor-specific single nucleotide variants that accurately distinguish tumor DNA from wild type DNA are promising targets. The reliable detection and quantification of these single-base DNA variants is technically challenging. Currently, a variety of techniques is applied, with no apparent "gold standard". Here we present a novel qPCR protocol that meets the conditions of extreme sensitivity and specificity that are required for detection and quantification of tumor DNA. By consecutive application of two polymerases, one of them designed for extreme base-specificity, the method reaches unprecedented sensitivity and specificity. Three qPCR assays were tested with spike-in experiments, specific for point mutations BRAF V600E, PTEN T167A and NRAS Q61L of melanoma cell lines. It was possible to detect down to one copy of tumor DNA per reaction (Poisson distribution, at a background of up to 200 000 wild type DNAs. To prove its clinical applicability, the method was successfully tested on a small cohort of BRAF V600E positive melanoma patients.

  20. Green Transformational Leadership and Green Performance: The Mediation Effects of Green Mindfulness and Green Self-Efficacy

    OpenAIRE

    Yu-Shan Chen; Ching-Hsun Chang; Yu-Hsien Lin

    2014-01-01

    No prior literature explores the influence of green transformational leadership on green performance, thus, this study develops a novel research framework to fill the research gap. This study investigates the influence of green transformational leadership on green performance and discusses the mediation effects of green mindfulness and green self-efficacy by means of structural equation modeling (SEM). The results indicate that green transformational leadership positively influences green min...

  1. Wet-lab tested microRNA assays for qPCR studies with SYBR®Green and DNA primers in pig tissues

    DEFF Research Database (Denmark)

    Mentzel, Caroline M. Junker; Skovgaard, Kerstin; Córdoba, Sarai

    2014-01-01

    MicroRNAs are key post-transcriptional regulators of gene expression that are involved in several biological processes including those that mediate disease pathophysiology. Hence, quantifying microRNA expression levels can provide important and novel insights into disease biology. In recent years...

  2. Green Thunderstorms Observed.

    Science.gov (United States)

    Gallagher, Frank W., III; Beasley, William H.; Bohren, Craig F.

    1996-12-01

    Green thunderstorms have been observed from time to time in association with deep convection or severe weather events. Often the green coloration has been attributed to hail or to reflections of light from green foliage on the ground. Some skeptics who have not personally observed a green thunderstorm do not believe that green thunderstorms exist. They suggest that the green storms may be fabrications by excited observers. The authors have demonstrated the existence of green thunderstorms objectively using a spectrophotometer. During the spring and summer of 1995 the authors observed numerous storms and recorded hundreds of spectra of the light emanating corn these storms. It was found that the subjective judgment of colors can vary somewhat between observers, but the variation is usually in the shade of green. The authors recorded spectra of green and nongreen thunderstorms and recorded spectral measurements as a storm changed its appearance from dark blue to a bluish green. The change in color is gradual when observed from a stationary position. Also, as the light from a storm becomes greener, the luminance decreases. The authors also observed and recorded the spectrum of a thunderstorm during a period of several hours as they flew in an aircraft close to a supercell that appeared somewhat green. The authors' observations refute the ground reflection hypothesis and raise questions about explanations that require the presence of hail.

  3. Properties of the reverse transcription reaction in mRNA quantification

    DEFF Research Database (Denmark)

    Ståhlberg, Anders; Håkansson, Joakim; Xian, Xiaojie

    2004-01-01

    BACKGROUND: In most measurements of gene expression, mRNA is first reverse-transcribed into cDNA. We studied the reverse transcription reaction and its consequences for quantitative measurements of gene expression. METHODS: We used SYBR green I-based quantitative real-time PCR (QPCR) to measure...... the properties of reverse transcription reaction for the beta-tubulin, glyceraldehyde-3-phosphate dehydrogenase, Glut2, CaV1D, and insulin II genes, using random hexamers, oligo(dT), and gene-specific reverse transcription primers. RESULTS: Experimental variation in reverse transcription-QPCR (RT......-QPCR) was mainly attributable to the reverse transcription step. Reverse transcription efficiency depended on priming strategy, and the dependence was different for the five genes studied. Reverse transcription yields also depended on total RNA concentration. CONCLUSIONS: RT-QPCR gene expression measurements...

  4. Latent class analysis of real time qPCR and bacteriological culturing for the diagnosis of Streptococcus agalactiae in cow composite milk samples.

    Science.gov (United States)

    Holmøy, Ingrid H; Toft, Nils; Jørgensen, Hannah J; Mørk, Tormod; Sølverød, Liv; Nødtvedt, Ane

    2018-06-01

    Streptococcus agalactiae (S. agalactiae) has re-emerged as a mastitis pathogen among Norwegian dairy cows. The Norwegian cattle health services recommend that infected herds implement measures to eradicate S. agalactiae, this includes a screening of milk samples from all lactating cows. The performance of the qPCR-test currently in use for this purpose has not been evaluated under field conditions. The objective of this study was to estimate the sensitivity and specificity of the real-time qPCR assay in use in Norway (Mastitis 4 qPCR, DNA Diagnostics A/S, Risskov, Denmark) and compare it to conventional bacteriological culturing for detection of S. agalactiae in milk samples. Because none of these tests are considered a perfect reference test, the evaluation was performed using latent class models in a Bayesian analysis. Aseptically collected cow-composite milk samples from 578 cows belonging to 6 herds were cultured and tested by qPCR. While 37 (6.4%) samples were positive for S. agalactiae by bacteriological culture, 66 (11.4%) samples were positive by qPCR. The within-herd prevalence in the six herds, as estimated by the latent class models ranged from 7.7 to 50.8%. At the recommended cut-off (cycle threshold 37), the sensitivity of the qPCR was significantly higher at 95.3 (95% posterior probability interval [PPI] [84.2; 99.6]) than that of bacteriological culture at 58.2 (95% PPI [43.8; 74.4]). However, bacterial culture had a higher specificity of 99.7 (95% PPI [98.5; 100.0]) compared to the qPCR at 98.5 (95% PPI [94.6; 99.9]). The median estimated negative predictive values of qPCR was consistently higher than those of the BC at all estimated prevalences, and the superiority of the qPCR increased with increasing within-herd prevalence. The median positive predictive values of BC was in general higher than the estimates for the qPCR, however, at the highest prevalence the predictive ability of both tests were similar. Copyright © 2018 Elsevier B.V. All

  5. A method for quantitative analysis of standard and high-throughput qPCR expression data based on input sample quantity.

    Directory of Open Access Journals (Sweden)

    Mateusz G Adamski

    Full Text Available Over the past decade rapid advances have occurred in the understanding of RNA expression and its regulation. Quantitative polymerase chain reactions (qPCR have become the gold standard for quantifying gene expression. Microfluidic next generation, high throughput qPCR now permits the detection of transcript copy number in thousands of reactions simultaneously, dramatically increasing the sensitivity over standard qPCR. Here we present a gene expression analysis method applicable to both standard polymerase chain reactions (qPCR and high throughput qPCR. This technique is adjusted to the input sample quantity (e.g., the number of cells and is independent of control gene expression. It is efficiency-corrected and with the use of a universal reference sample (commercial complementary DNA (cDNA permits the normalization of results between different batches and between different instruments--regardless of potential differences in transcript amplification efficiency. Modifications of the input quantity method include (1 the achievement of absolute quantification and (2 a non-efficiency corrected analysis. When compared to other commonly used algorithms the input quantity method proved to be valid. This method is of particular value for clinical studies of whole blood and circulating leukocytes where cell counts are readily available.

  6. GREEN MANAGEMENT: THE REALITY OF BEING GREEN IN BUSINESS

    OpenAIRE

    Tran, Ben

    2009-01-01

    Green management and going green are not as clear cut and easy as hyped by the general media. While going ecologically green is indeed beneficial and appropriate, the process and procedure of becoming green is anything but easy. Firstly, turning green is largely not a legal requirement, but a voluntary process. Thus, even though LEED (which is by far the more publicly known green certification standard) governs the certification of the green management effort, it is not a compulsory condition...

  7. Touch-down reverse transcriptase-PCR detection of IgV(H) rearrangement and Sybr-Green-based real-time RT-PCR quantitation of minimal residual disease in patients with chronic lymphocytic leukemia

    Czech Academy of Sciences Publication Activity Database

    Peková, Soňa; Marková, J.; Pajer, Petr; Dvořák, Michal; Cetkovský, P.; Schwarz, J.

    2005-01-01

    Roč. 9, č. 1 (2005), s. 23-34 ISSN 1084-8592 Institutional research plan: CEZ:AV0Z50520514 Keywords : minimal residual disease * chronic lymphocytic leukaemia * IgV (H) rearrangement Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.562, year: 2003

  8. Direct Comparison of the Histidine-rich Protein-2 Enzyme-linked Immunosorbent Assay (HRP-2 ELISA) and Malaria SYBR Green I Fluorescence (MSF) Drug Sensitivity Tests in Plasmodium falciparum Reference Clones and Fresh ex vivo Field Isolates from Cambodia

    Science.gov (United States)

    2013-07-12

    assessment of antimalarial activity in vitro by a semiautomated microdilution technique. Antimicrob Agents Chemother 1979, 16:710–718. 3. Noedl H, Attlmayr B...40:685–691. 32. Hawley SR, Bray PG, Mungthin M, Atkinson JD, O’Neill PM, Ward SA: Relationship between antimalarial drug activity , accumulation, and...success rate when testing DHA, AS, MQ, QN, CQ, and PPQ activities . A “successful” IC50 assay result for each P. falciparum clinical isolate was defined as

  9. Green energy in Europe: selling green energy with green certificates

    International Nuclear Information System (INIS)

    Ouillet, L.

    2002-01-01

    Sales of green power products are booming in Europe: 50,000 customers in the United Kingdom, 775,000 in the Netherlands and 300,000 in Germany. Laws of physics are however formal: the way in which electricity flows within the grid does not allow suppliers to assure customers that they are directly receiving electricity produced exclusively from renewable energy sources. What are marketers selling their customers then? Laetitia Ouillet, Greenprices, takes a closer look and focuses on the potential of selling green energy in the forms of renewable energy certificates. (Author)

  10. The utility of optical detection system (qPCR) and bioinformatics methods in reference gene expression analysis

    Science.gov (United States)

    Skarzyńska, Agnieszka; Pawełkowicz, Magdalena; PlÄ der, Wojciech; Przybecki, Zbigniew

    2016-09-01

    Real-time quantitative polymerase chain reaction is consider as the most reliable method for gene expression studies. However, the expression of target gene could be misinterpreted due to improper normalization. Therefore, the crucial step for analysing of qPCR data is selection of suitable reference genes, which should be validated experimentally. In order to choice the gene with stable expression in the designed experiment, we performed reference gene expression analysis. In this study genes described in the literature and novel genes predicted as control genes, based on the in silico analysis of transcriptome data were used. Analysis with geNorm and NormFinder algorithms allow to create the ranking of candidate genes and indicate the best reference for flower morphogenesis study. According to the results, genes CACS and CYCL were characterised the most stable expression, but the least suitable genes were TUA and EF.

  11. Early detection of sugar beet pathogen Ramularia beticola in leaf and air samples using qPCR

    DEFF Research Database (Denmark)

    Wieczorek, Thies Marten; Jørgensen, Lise Nistrup; Hansen, Anne Lisbet

    2014-01-01

    A quantitative PCR method (qPCR) was developed for the detection and quantification of Ramularia beticola causing Ramularia leaf spot in sugar beet. R. beticola specific primers were designed based on the internal transcribed spacer region 2 (ITS2). The assay was applied on DNA extracted from...... spores trapped on tape from Burkard spore traps placed in an artificially inoculated sugar beet field trial and in two sugar beet fields with natural infections. R. beticola DNA was detected at variable amounts in the air samples 14 to 16 days prior to first visible symptoms. R. beticola DNA was detected...... in air samples from fields with natural infection at significant and increasing levels from development of the first symptoms, indicating that spore production within the crop plays a major role in the epidemic development of the disease. Sugar beet leaves sampled from the inoculated field trial were...

  12. Analysis of variation in virulence of Beauveria bassiana against insect pests of pigeonpea using qPCR.

    Science.gov (United States)

    Senthilraja, Govindasamy; Anand, Theerthagiri; Mohankumar, Subbarayalu; Raguchander, Thiruvengadam; Samiyappan, Ramasamy

    2018-03-01

    Beauveria bassiana is a broad spectrum microbial bioagent used for the control of agriculturally important insect pests. However, in our experiments, two virulent isolates of B. bassiana (B2 and B10) showed specific preference toward Maruca vitrata and Helicoverpa armigera of pigeonpea. To better understand this feature, we developed a qPCR assay to quantify the chitinase (virulence factor) of B. bassiana during the infection process. Isolates of B. bassiana were grown on insect cuticle amended medium and minimal medium (without insect cuticle) to assess the induction of chitinase. Our results revealed a positive correlation between expression of chitinase by B. bassiana and the substrates (with or without cuticles of M. vitrata and H. armigera) used. This study showcases the methodology to quantify the chitinase and analysis of variation in virulence of B. bassiana (B2 and B10) against M. vitrata and H. armigera. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Determining the 95% limit of detection for waterborne pathogen analyses from primary concentration to qPCR

    Science.gov (United States)

    Stokdyk, Joel P.; Firnstahl, Aaron; Spencer, Susan K.; Burch, Tucker R; Borchardt, Mark A.

    2016-01-01

    The limit of detection (LOD) for qPCR-based analyses is not consistently defined or determined in studies on waterborne pathogens. Moreover, the LODs reported often reflect the qPCR assay alone rather than the entire sample process. Our objective was to develop an approach to determine the 95% LOD (lowest concentration at which 95% of positive samples are detected) for the entire process of waterborne pathogen detection. We began by spiking the lowest concentration that was consistently positive at the qPCR step (based on its standard curve) into each procedural step working backwards (i.e., extraction, secondary concentration, primary concentration), which established a concentration that was detectable following losses of the pathogen from processing. Using the fraction of positive replicates (n = 10) at this concentration, we selected and analyzed a second, and then third, concentration. If the fraction of positive replicates equaled 1 or 0 for two concentrations, we selected another. We calculated the LOD using probit analysis. To demonstrate our approach we determined the 95% LOD for Salmonella enterica serovar Typhimurium, adenovirus 41, and vaccine-derived poliovirus Sabin 3, which were 11, 12, and 6 genomic copies (gc) per reaction (rxn), respectively (equivalent to 1.3, 1.5, and 4.0 gc L−1 assuming the 1500 L tap-water sample volume prescribed in EPA Method 1615). This approach limited the number of analyses required and was amenable to testing multiple genetic targets simultaneously (i.e., spiking a single sample with multiple microorganisms). An LOD determined this way can facilitate study design, guide the number of required technical replicates, aid method evaluation, and inform data interpretation.

  14. Determination of the Physical Status (Episomal/Integral of HPV by qPCR in Esophageal Squamous Cell Carcinoma

    Directory of Open Access Journals (Sweden)

    Fariborz Soheili

    2017-03-01

    Full Text Available Background: In cervical cancer, the carcinogenic mechanism of human papillomavirus (HPV occurs through the integration of viral DNA into the host genome. This process initiates with a disruption in the E2 open reading frame (ORF of the viral genome. Disruption of E2 ORF results in an increased expression of the viral oncoproteins, E6 and E7, by removal of E2 suppression effect on their promoters. E6 and E7 interfere with the normal cell cycle by degrading the p53 and pRb tumor suppressor proteins, respectively. Objectives: The objective of this study was to determine the physical status (episomal/integral of HPV genome in esophageal squamous cell carcinoma (ESCC. Materials and Methods: The rate of copy numbers of E2 and E6 genes in HPV-18 and HPV-16 positive samples were analyzed by quantitative polymerase chain reaction (qPCR in order to assess the physical status (episomal/integral of HPV. DNA extracts from HeLa cell line were used as the positive control. Results: The E2 gene was detected in 1 sample, co-infected with HPV-16 and HPV-18. While, E6 gene was detected in all 11 HPV positive samples. The qPCR analysis showed the presence of integrated form of viral DNA in all HPV positive samples and only 1 mixed episomal-integrated form was detected. Conclusion: The presence of integrated forms of high risk HPV-16 and HPV-18 genomes might reflect a crucial process towards malignant transformation of ESCC.

  15. Seawater detection and biological assessments regarding transmission of the oyster parasite Mikrocytos mackini using qPCR.

    Science.gov (United States)

    Polinski, Mark P; Meyer, Gary R; Lowe, Geoffrey J; Abbott, Cathryn L

    2017-10-18

    Mikrocytos mackini is an intracellular parasite of oysters and causative agent of Denman Island disease in Pacific oysters Crassostrea gigas. Although M. mackini has been investigated for decades, its natural mode of transmission, mechanism for host entry, and environmental stability are largely unknown. We explored these biological characteristics of M. mackini using a recently described quantitative PCR (qPCR) assay. We detected M. mackini in the flow-through tank water of experimentally infected oysters and during disease remission in host tissues following 6 wk of elevated water temperature. Waterborne exposure of oysters to M. mackini further confirmed the potential for extracellular seawater transmission of this parasite and also identified host gill to have the highest early and continued prevalence for M. mackini DNA compared to stomach, mantle, labial palps, or adductor muscle samples. However, infections following waterborne challenge were slow to develop despite a substantial exposure (>106 M. mackini l-1 for 24 h), and further investigation demonstrated that M. mackini occurrence and infectivity severely declined following extracellular seawater incubation of more than 24 h. This study demonstrates a potential for using qPCR to monitor M. mackini in wild or farmed oyster populations during periods of disease remission or from environmental seawater samples. This work also suggests that gill tissues may provide a primary site for waterborne entry and possibly shedding of M. mackini in oysters. Further, although extracellular seawater transmission of M. mackini was possible, poor environmental stability and infection efficiency likely restricts the geographic transmission of M. mackini between oysters in natural environs and may help to explain localized areas of infection.

  16. Green growth in fisheries

    DEFF Research Database (Denmark)

    Nielsen, Max; Ravensbeck, Lars; Nielsen, Rasmus

    2014-01-01

    harming the environment. Fishery is an environment-dependent sector and it has been argued that there is no potential for green growth in the sector owing to global overexploitation, leaving no scope for production growth. The purpose of this paper is to explain what green growth is and to develop......Climate change and economic growth have gained a substantial amount of attention over the last decade. Hence, in order to unite the two fields of interest, the concept of green growth has evolved. The concept of green growth focuses on how to achieve growth in environment-dependent sectors, without...... a conceptual framework. Furthermore, the aim is to show that a large green growth potential actually exists in fisheries and to show how this potential can be achieved. The potential green growth appears as value-added instead of production growth. The potential can be achieved by reducing overcapacity...

  17. Rapid Identification and Quantification of Aureococcus anophagefferens by qPCR Method (Taqman) in the Qinhuangdao Coastal Area: A Region for Recurrent Brown Tide Breakout in China.

    Science.gov (United States)

    Wang, Li-Ping; Lei, Kun

    2016-12-01

    Since 2009, Aureococcus anophagefferens has caused brown tide to occur recurrently in Qinhuangdao coastal area, China. Because the algal cells of A. anophagefferens are so tiny (~3 µm) that it is very hard to identify exactly under a microscope for natural water samples, it is very urgent to develop a method for efficient and continuous monitoring. Here specific primers and Taqman probe are designed to develop a real-time quantitative PCR (qPCR) method for identification and quantification continually. The algal community and cell abundance of A. anophagefferens in the study area (E 119°20'-119°50' and N 39°30'-39°50') from April to October in 2013 are detected by pyrosequencing, and are used to validate the specification and precision of qPCR method for natural samples. Both pyrosequencing and qPCR shows that the targeted cells are present only in May, June and July, and the cell abundance are July > June > May. Although there are various algal species including dinoflagellata, diatom, Cryptomonadales, Chrysophyceae and Chlorophyta living in the natural seawater simultaneously, no disturbance happens to qPCR method. This qPCR method could detect as few as 10 targeted cells, indicating it is able to detect the algal cells at pre-bloom levels. Therefore, qPCR with Taqman probe provides a powerful and sensitive method to monitor the brown tide continually in Qinhuangdao coastal area, China. The results provide a necessary technology support for forecasting the brown tide initiation, in China.

  18. Green electricity buyer's guide

    International Nuclear Information System (INIS)

    Kelly, B.; Klein, S.; Olivastri, B.

    2002-06-01

    The electricity produced in whole or in large part from renewable energy sources like wind, small hydro electricity and solar energy, is generally referred to as green electricity. The authors designed this buyer's guide to assist customers in their understanding of green electricity, as the customers can now choose their electricity supplier. The considerations and steps involved in the purchasse of green electricity are identified, and advice is provided on ways to maximize the benefits from the purchase of green electricity. In Alberta and Ontario, customers have access to a competitive electricity market. The emphasis when developing this guide was placed firmly on the large buyers, as they can have enormous positive influence on the new market for green electricity. The first chapter of the document provides general information on green electricity. In chapter two, the authors explore the opportunity for environmental leadership. Chapter three reviews the basics of green electricity, which provides the link to chapter four dealing with the creation of a policy. Purchasing green electricity is dealt with in Chapter five, and maximizing the benefits of green electricity are examined in Chapter Six. 24 refs., 3 tabs

  19. GREEN PACKAGING, GREEN PRODUCT, GREEN ADVERTISING, PERSEPSI, DAN MINAT BELI KONSUMEN

    OpenAIRE

    Imam Santoso; Rengganis Fitriani

    2016-01-01

    Environmental problems become one of the strategic issues in achieving global competitiveness. One of the issues is products that are made from environmental friendly materials or known as green product. Furthermore, in green products marketing, the company also uses green packaging and green advertising concept. This study aimed to analyze the effect of green packaging, green products, and green advertising on consumer perception and purchasing intention. The study was conducted in Ketawangg...

  20. Customers’ Intention to Use Green Products: the Impact of Green Brand Dimensions and Green Perceived Value

    Directory of Open Access Journals (Sweden)

    Doszhanov Aibek

    2015-01-01

    Full Text Available This study aimed to identify the relationships between green brand dimension (green brand awareness, green brand image, and green brand trust, green perceived value and customer’s intention to use green products. Data was collected through structured survey questionnaire from 384 customers of three hypermarkets in Kuala-Lumpur. Data was analyzed based on multiple regression analysis. The results indicate that there are significant relationships between green brand awareness, green brand trust, green perceived value, and customer’s intention to use green products. However, green brand image was not found to have significant relationship with customer’s intention to use green products. The discussion presented suggestions for marketers and researchers interested in green branding.

  1. Greening the Future

    Science.gov (United States)

    Williamson, Norma Velia

    2011-01-01

    Because educators vicariously touch the future through their students, the author believes that they sometimes have the uncanny ability to see the future. One common future forecast is the phenomenal growth of green jobs in the emerging green economy, leading to the creation of the "Reach of the Sun" Solar Energy Academy at La Mirada…

  2. Green Buildings and Health.

    Science.gov (United States)

    Allen, Joseph G; MacNaughton, Piers; Laurent, Jose Guillermo Cedeno; Flanigan, Skye S; Eitland, Erika Sita; Spengler, John D

    2015-09-01

    Green building design is becoming broadly adopted, with one green building standard reporting over 3.5 billion square feet certified to date. By definition, green buildings focus on minimizing impacts to the environment through reductions in energy usage, water usage, and minimizing environmental disturbances from the building site. Also by definition, but perhaps less widely recognized, green buildings aim to improve human health through design of healthy indoor environments. The benefits related to reduced energy and water consumption are well-documented, but the potential human health benefits of green buildings are only recently being investigated. The objective of our review was to examine the state of evidence on green building design as it specifically relates to indoor environmental quality and human health. Overall, the initial scientific evidence indicates better indoor environmental quality in green buildings versus non-green buildings, with direct benefits to human health for occupants of those buildings. A limitation of much of the research to date is the reliance on indirect, lagging and subjective measures of health. To address this, we propose a framework for identifying direct, objective and leading "Health Performance Indicators" for use in future studies of buildings and health.

  3. Green product innovation strategy

    NARCIS (Netherlands)

    Driessen, P.H.

    2005-01-01

    Over the last decades, companies have started to incorporate green issues in product innovation strategies. This dissertation studies green product innovation strategy, its antecedents and its outcomes. A three-stage approach is followed. In the first stage, the topic is explored and a preliminary

  4. Green for rarity

    International Nuclear Information System (INIS)

    Raal, F.A.; Robinson, D.N.

    1980-01-01

    Green diamonds once recovered from Witwatersrand gold/uranium deposits, are now a thing of the past with the modernisation of extraction metallurgy methods. The green colouration has been shown to be due to radiation from uranium present in the ore

  5. Green Building Research Laboratory

    Energy Technology Data Exchange (ETDEWEB)

    Sailor, David Jean [Portland State Univ., Portland, OR (United States)

    2013-12-29

    This project provided support to the Green Building Research Laboratory at Portland State University (PSU) so it could work with researchers and industry to solve technical problems for the benefit of the green building industry. It also helped to facilitate the development of PSU’s undergraduate and graduate-level training in building science across the curriculum.

  6. Green Cleaning Label Power

    Science.gov (United States)

    Balek, Bill

    2012-01-01

    Green cleaning plays a significant and supportive role in helping education institutions meet their sustainability goals. However, identifying cleaning products, supplies and equipment that truly are environmentally preferable can be daunting. The marketplace is inundated with products and services purporting to be "green" or environmentally…

  7. Introduction: Experimental Green Strategies

    DEFF Research Database (Denmark)

    Peters, Terri

    2011-01-01

    Defining new ways in which archietcts are responding to the challenge of creating sustainable architecture , Experimental Green Strategies present a state of the art in applied ecological architectural research.......Defining new ways in which archietcts are responding to the challenge of creating sustainable architecture , Experimental Green Strategies present a state of the art in applied ecological architectural research....

  8. Green heterogeneous wireless networks

    CERN Document Server

    Ismail, Muhammad; Nee, Hans-Peter; Qaraqe, Khalid A; Serpedin, Erchin

    2016-01-01

    This book focuses on the emerging research topic "green (energy efficient) wireless networks" which has drawn huge attention recently from both academia and industry. This topic is highly motivated due to important environmental, financial, and quality-of-experience (QoE) considerations. Specifically, the high energy consumption of the wireless networks manifests in approximately 2% of all CO2 emissions worldwide. This book presents the authors’ visions and solutions for deployment of energy efficient (green) heterogeneous wireless communication networks. The book consists of three major parts. The first part provides an introduction to the "green networks" concept, the second part targets the green multi-homing resource allocation problem, and the third chapter presents a novel deployment of device-to-device (D2D) communications and its successful integration in Heterogeneous Networks (HetNets). The book is novel in that it specifically targets green networking in a heterogeneous wireless medium, which re...

  9. Building the green way.

    Science.gov (United States)

    Lockwood, Charles

    2006-06-01

    Just five or six years ago, the term "green building" evoked visions of barefoot, tie-dyed, granola-munching denizens. There's been a large shift in perception. Of course, green buildings are still known for conserving natural resources by, for example, minimizing on-site grading, using alternative materials, and recycling construction waste. But people now see the financial advantages as well. Well-designed green buildings yield lower utility costs, greater employee productivity, less absenteeism, and stronger attraction and retention of workers than standard buildings do. Green materials, mechanical systems, and furnishings have become more widely available and considerably less expensive than they used to be-often cheaper than their standard counterparts. So building green is no longer a pricey experiment; just about any company can do it on a standard budget by following the ten rules outlined by the author. Reliable building-rating systems like the U.S. Green Building Council's rigorous Leadership in Energy and Environmental Design (LEED) program have done much to underscore the benefits of green construction. LEED evaluates buildings and awards points in several areas, such as water efficiency and indoor environmental quality. Other rating programs include the UK's BREEAM (Building Research Establishment's Environmental Assessment Method) and Australia's Green Star. Green construction is not simply getting more respect; it is rapidly becoming a necessity as corporations push it fully into the mainstream over the next five to ten years. In fact, the author says, the owners of standard buildings face massive obsolescence. To avoid this problem, they should carry out green renovations. Corporations no longer have an excuse for eschewing environmental and economic sustainability. They have at their disposal tools proven to lower overhead costs, improve productivity, and strengthen the bottom line.

  10. Standardization of enterococci density estimates by EPA qPCR methods and comparison of beach action value exceedances in river waters with culture methods

    Science.gov (United States)

    The U.S.EPA has published recommendations for calibrator cell equivalent (CCE) densities of enterococci in recreational waters determined by a qPCR method in its 2012 Recreational Water Quality Criteria (RWQC). The CCE quantification unit stems from the calibration model used to ...

  11. Application of real-time PCR (qPCR) for characterization of microbial populations and type of milk in dairy food products.

    Science.gov (United States)

    Agrimonti, Caterina; Bottari, Benedetta; Sardaro, Maria Luisa Savo; Marmiroli, Nelson

    2017-09-08

    Dairy foods represent an important sector of the food market for their nutritional qualities and their organoleptic characteristics, which are often linked to tradition and to region. These products are typically protected by labels such as PDO (Protected Designation of Origin) and PGI (Protected Geographical Indication). Real-time PCR (qPCR) is a fundamental tool in "Food Genomics;" a discipline concerned with the residual DNA in food, which, alongside traditional physical and chemical methods, is frequently used to determine product safety, quality and authenticity. Compared to conventional or "end-point" PCR, qPCR incorporates continuous monitoring of reaction progress, thereby enabling quantification of target DNA. This review describes qPCR applications to the analysis of microbiota, and to the identification of the animal species source of milk from which dairy products have been made. These are important aspects for ensuring safety and authenticity. The various applications of qPCR are discussed, as well as advantages and disadvantages in comparison with other analytical methods.

  12. Human-associated fecal qPCR measurements and predicted risk of gastrointestinal illness in recreational waters contaminated with raw sewage

    Science.gov (United States)

    We used quantitative microbial risk assessment (QMRA) to estimate the risk of gastrointestinal (GI) illness associated with swimming in recreational waters containing different concentrations of human-associated fecal qPCR markers from raw sewage– HF183 and HumM2. The volume/volu...

  13. Clarkesville Green Infrastructure Implementation Strategy

    Science.gov (United States)

    The report outlines the 2012 technical assistance for Clarkesville, GA to develop a Green Infrastructure Implementation Strategy, which provides the basic building blocks for a green infrastructure plan:

  14. The sensitivity and efficacy method of PIK3CA exon 9 E545A as a high diagnostic accuracy in breast cancer

    Directory of Open Access Journals (Sweden)

    Desriani

    2018-06-01

    Full Text Available The phosphatidylinositol 3-kinases (PIK3s are lipid kinases. Mutation in the exon 9 and exon 20 determined as a predictive factor in anti-HER-2 therapy. In some countries, such as Singapore, China, and Peru, PIK3CA exon 9 E545A was reported to produce the highest rate of mutation. In this research, we developed and optimized PIK3CA exon 9 E545A detection methods with intercalating dye SYBR Green I based on the Tm Shift approach by using prepared recombinant plasmid pGEMT-easy PIK3CA exon 9 and PIK3CA exon 9 E545A. Recombinant plasmid was used due to the limited number of samples. Methods: Recombinant plasmid was prepared based on manufactured procedures, and this process was then followed by Tm prediction with Poland software, Tm Shift SYBR Green I development, and its characterization (reproducibility, repeatability, sensitivity, qPCR efficiency, and qPCR amplification, respectively. Result: A method for PIK3CA E545A detection based on TM shift SYBR Green I has been successfully developed. The melting temperature for PIK3CA exon 9 was 78.1 ± 0.1 °C, while that for PIK3CA exon E545A was 80.20 °C. The Tm of mutant was the same as that predicted using Polland Software. The reproducibility of the methods was high, with the coefficient values for inter and intra assays were below 10% with a high sensitivity at 1%, while R2 0.99 and PCR efficiency was 97.75%. Conclusion: The results presented here demonstrate that the PIK3CA exon 9 E545A detection method has a good sensitivity and efficacy assay, which proves that the method has a high diagnostic accuracy in breast cancer. Keywords: SYBR Green I, PIK3CA E545A, Breast cancer, Real time PCR, Recombinant plasmid

  15. Selling the green dream

    International Nuclear Information System (INIS)

    Wood, E.

    2005-01-01

    The article discusses the marketing and sales of energy generated from renewable energy sources. To purchase environmental energy in the USA, the consumer need do no more than tick a box on a sheet of paper. But, it is not just households that opt for green energy: businesses are also willing customers. A factor in the success in selling green energy to big business is that the retail price of wind power can be held constant over periods of several years, whereas fossil fuel prices can fluctuate wildly. Details of sources and sales of the top ten companies selling green energy are given

  16. Manufacturing Green Consensus

    DEFF Research Database (Denmark)

    Gulsrud, Natalie Marie; Ooi, Can Seng

    2014-01-01

    In an increasingly global economy, being green, or having an environmentally sustainbale place brand, provides a competitive advantage. Singapore, long known as the ``garden city´´ has been a leader in green city imaging since the founding of the equatorial city-state, contributing, in large part...... to the city’s profile as the economic giant of Southeast Asia. Using a political ecology lens, the paper aims to uncover the contested socio-economic narratives of green city imaging by examining the evolution of the garden city branding scheme since Singapore’s independence in 1959. Results show...

  17. About green political parties

    Directory of Open Access Journals (Sweden)

    Orlović Slobodan P.

    2015-01-01

    Full Text Available In this work the author refers to some legal and political questions in connection with green political parties. Those questions cover: the ideology of green political parties, their number and influence, both in general and in Serbia. The first part of work is generally speaking about political parties - their definition, ideology, role and action. Main thesis in this work is that green political parties, by their appearance, were something new on the political scene. But quickly, because of objective and subjective reasons, they were changing original ideas and were beginning to resemble to all other political parties. In this way, they lost their vanguard and political alternativeness.

  18. Validation and comparison of reference genes for qPCR normalization of celery (Apium graveolens at different development stages

    Directory of Open Access Journals (Sweden)

    Meng-Yao eLi

    2016-03-01

    Full Text Available A suitable reference gene is an important prerequisite for guarantying accurate and reliable results in qPCR analysis. Celery is one of the representative vegetable in Apiaceae and is widely cultivated and consumed in the world. However, no reports have been previously published concerning reference genes in celery. In this study, the expression stabilities of nine candidate reference genes in leaf blade and petiole at different development stages were evaluated using three statistics algorithms geNorm, NormFinder, and BestKeeper. Our results showed that TUB-B, TUB-A, and UBC were the most reference genes among all tested samples. GAPDH represented the maximum stability for most individual sample, while the UBQ displayed the minimum stability. To further validate the stability of reference genes, the expression pattern of AgAP2-2 was calculated by using the selected genes for normalization. In addition, the expression patterns of several development-related genes were studied using the selected reference gene. Our results will be beneficial for further studies on gene transcription in celery.

  19. A rapid Q-PCR titration protocol for adenovirus and helper-dependent adenovirus vectors that produces biologically relevant results

    Science.gov (United States)

    Gallaher, Sean D.; Berk, Arnold J.

    2013-01-01

    Adenoviruses are employed in the study of cellular processes and as expression vectors used in gene therapy. The success and reproducibility of these studies is dependent in part on having accurate and meaningful titers of replication competent and helper-dependent adenovirus stocks, which is problematic due to the use of varied and divergent titration protocols. Physical titration methods, which quantify the total number of viral particles, are used by many, but are poor at estimating activity. Biological titration methods, such as plaque assays, are more biologically relevant, but are time consuming and not applicable to helper-dependent gene therapy vectors. To address this, a protocol was developed called “infectious genome titration” in which viral DNA is isolated from the nuclei of cells ~3 h post-infection, and then quantified by Q-PCR. This approach ensures that only biologically active virions are counted as part of the titer determination. This approach is rapid, robust, sensitive, reproducible, and applicable to all forms of adenovirus. Unlike other Q-PCR-based methods, titers determined by this protocol are well correlated with biological activity. PMID:23624118

  20. In silico prediction and qPCR validation of novel sRNAs in Propionibacterium acnes KPA171202

    Directory of Open Access Journals (Sweden)

    Praveen P. Balgir

    2016-06-01

    Full Text Available Propionibacterium acnes is an anaerobic, Gram-positive, opportunistic pathogen known to be involved in a wide variety of diseases ranging from mild acne to prostate cancer. Bacterial small non-coding RNAs are novel regulators of gene expression and are known to be involved in, virulence, pathogenesis, stress tolerance and adaptation to environmental changes in bacteria. The present study was undertaken keeping in view the lack of predicted sRNAs of P. acnes KPA171202 in databases. This report represents the first attempt to identify sRNAs in P. acnes KPA171202. A total of eight potential candidate sRNAs were predicted using SIPHT, one was found to have a Rfam homolog and seven were novel. Out of these seven predicted sRNAs, five were validated by reverse transcriptase-polymerase chain reaction (RT-PCR and sequencing. The expression of these sRNAs was quantified in different growth phases by qPCR (quantitative PCR. They were found to be expressed in both exponential and stationary stages of growth but with maximum expression in stationary phase which points to a regulatory role for them. Further investigation of their targets and regulatory functions is in progress.

  1. Sustainable green urban planning: the Green Credit Tool

    NARCIS (Netherlands)

    Cilliers, E.J.; Diemont, E.; Stobbelaar, D.J.; Timmermans, W.

    2010-01-01

    Purpose – The Green Credit Tool is evaluated as a method to quantify the value of green-spaces and to determine how these green-space-values can be replaced or compensated for within urban spatial planning projects. Design/methodology/approach – Amersfoort Local Municipality created the Green Credit

  2. Green Supplier Selection Criteria

    DEFF Research Database (Denmark)

    Nielsen, Izabela Ewa; Banaeian, Narges; Golinska, Paulina

    2014-01-01

    Green supplier selection (GSS) criteria arise from an organization inclination to respond to any existing trends in environmental issues related to business management and processes, so GSS is integrating environmental thinking into conventional supplier selection. This research is designed...... to determine prevalent general and environmental supplier selection criteria and develop a framework which can help decision makers to determine and prioritize suitable green supplier selection criteria (general and environmental). In this research we considered several parameters (evaluation objectives......) to establish suitable criteria for GSS such as their production type, requirements, policy and objectives instead of applying common criteria. At first a comprehensive and deep review on prevalent and green supplier selection literatures performed. Then several evaluation objectives defined to assess the green...

  3. Green Building Standards

    Science.gov (United States)

    Many organizations have developed model codes or rating systems that communities may use to develop green building programs or revise building ordinances. Some of the major options are listed on this page.

  4. Introduction: Green Building Handbook

    CSIR Research Space (South Africa)

    Van Wyk, Llewellyn V

    2010-08-01

    Full Text Available By recognising the specific environmental challenges facing South Africa, mindful of the government‘s commitment to reducing South Africa‘s Greenhouse gas emissions, and acknowledging the need to build social cohesion, the Green Building Handbook...

  5. Green certificates causing inconvenience?

    International Nuclear Information System (INIS)

    Torgersen, Lasse

    2002-01-01

    From early 2002, producers of green energy in selected countries have been able to benefit from generous financial support in the Netherlands. Thus, there has been increased sale of green certificates from Norway and Sweden. But the condition that physical energy delivery should accompany the certificates has caused a marked rise in the price of energy in transit through Germany to the Netherlands. This article discusses the green certificate concept and the experience gained from the Netherlands. One conclusion is that if large-scale trade with green certificates is introduced in Europe without the condition of accompanying energy delivery, then producers of hydro-electric power in Norway and Sweden may be the losers

  6. Green space as classroom

    DEFF Research Database (Denmark)

    Bentsen, Peter; Schipperijn, Jasper; Jensen, Frank Søndergaard

    2013-01-01

    More and more Danish teachers have started introducing curriculum-based outdoor learning as a weekly or biweekly ‘outdoor school’ day for school children. This move towards schooling in non-classroom spaces presents a challenge for green space managers. Basic managerial knowledge related to what......, who, when and where has thus far only been supported by anecdotal evidence, but seems fundamental to the decision-making of a range of green space providers. The present study aims to describe, characterise and discuss outdoor teachers’ use, preferences and ecostrategies in relation to green space....... A nationwide survey was conducted among Danish teachers practising outdoor teaching (107 respondents), and it showed that a majority used and preferred forest areas. The outdoor teachers used mainly school grounds and local green space for their outdoor teaching with a majority using the same place or mostly...

  7. Green by Default

    DEFF Research Database (Denmark)

    Sunstein, Cass R.; Reisch, Lucia

    2013-01-01

    The article offers information on the two sources of energy including green energy and gray energy. It discusses several facts which includes lower levels of greenhouse gases and conventional pollutants, relationship between economic incentives and underlying preferences and potential effects...

  8. Green Sturgeon Acoustic Monitoring

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This database is used to hold tracking data for green sturgeon tagged in Central California. The data collection began in late 2002 and is ongoing.

  9. Green Turtle Critical Habitat

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These data represent the critical habitat for green turtle as designated by Federal Register Vol. 63, No. 46701, September 2, 1998, Rules and Regulations.

  10. Green Turtle Trophic Ecology

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — SWFSC is currently conducting a study of green sea turtle (Chelonia mydas) trophic ecology in the eastern Pacific. Tissue samples and stable carbon and stable...

  11. Decon Green (trademark)

    National Research Council Canada - National Science Library

    Wagner, George W; Procell, Lawrence R; Henderson, Vikki D; Sorrick, David C; Hess, Zoe A; Gehring, David G; Brickhouse, Mark D

    2004-01-01

    ...; it affords the broad-spectrum decontamination of chemical and biological warfare agents. Composed entirely of ingredients commonly found in cosmetics, detergents, laundry boosters, and vitamins, Decon Green (trademark...

  12. Superfund Green Remediation

    Science.gov (United States)

    Green remediation is the practice of considering all environmental effects of site cleanup and incorporating options – like the use of renewable energy resources – to maximize the environmental benefits of cleanups.

  13. ADVANTAGES OF GREEN TECHNOLOGY

    OpenAIRE

    Ghanshyam Das Soni

    2017-01-01

    Technology is application of knowledge to practical requirements. Green technologies encompass various aspects of technology which help us reduce the human impact on the environment and create ways of sustainable development. Social equitability, economic feasibility and sustainability are the key parameters for green technologies. Today the environment is racing towards the tipping point at which we would have done permanent irreversible damage to the planet earth. Our current actions are pu...

  14. What is a green building?

    NARCIS (Netherlands)

    Vreenegoor, R.C.P.; Krikke, T.; Mierlo, van B.P.; Pluijm, van der W.M.P.; Poortvliet, R.; Hensen, J.L.M.; Loomans, M.G.L.C.

    2009-01-01

    What is a green building? A large amount of definitions and green rating tools prove that an exact definition is still a point of discussion. To research the differences between green rating tools, four different buildings are assessed with: EPN, BREEAM, LEED, GreenCalc+ and EcoQuantum. These tools

  15. The Influence of Proactive Green Innovation and Reactive Green Innovation on Green Product Development Performance: The Mediation Role of Green Creativity

    Directory of Open Access Journals (Sweden)

    Yu-Shan Chen

    2016-09-01

    Full Text Available This study fills the research gap in the exploration of the relationships between both proactive and reactive green innovations and green product development performance, and examines the mediating effect of green creativity. Structural equation modeling (SEM is utilized to test the hypotheses. From the sample of 146 valid respondents, the results show that proactive green innovation positively affects green creativity and green product development performance, and green creativity positively affects green product development performance. In addition, our findings also indicate that the relationship between proactive green innovation and green product development performance is partially mediated by green creativity. Accordingly, green creativity plays a critical role for companies to achieve a great green product development performance. However, reactive green innovation does not significantly influence green creativity and green product development performance. Companies should develop proactive green innovation rather than reactive green innovation in order to enhance their green creativity and increase their product development performance.

  16. A performance study on three qPCR quantification kits and their compatibilities with the 6-dye DNA profiling systems.

    Science.gov (United States)

    Lin, Sze-Wah; Li, Christina; Ip, Stephen C Y

    2018-03-01

    DNA quantification plays an integral role in forensic DNA profiling. Not only does it estimate the total amount of amplifiable human autosomal and male DNA to ensure optimal amplification of target DNA for subsequent analysis, but also assesses the extraction efficiency and purity of the DNA extract. Latest DNA quantification systems even offer an estimate for the degree of DNA degradation in a sample. Here, we report the performance of three new generation qPCR kits, namely Investigator ® Quantiplex HYres Kit from QIAGEN, Quantifiler ® Trio DNA Quantification Kit from Applied Biosystems™, and PowerQuant ® System from Promega, and their compatibilities with three 6-dye DNA profiling systems. Our results have demonstrated that all three kits generate standard curves with satisfactory consistency and reproducibility, and are capable of screening out traces of male DNA in the presence of 30-fold excess of female DNA. They also exhibit a higher tolerance to PCR inhibition than Quantifiler ® Human DNA Quantification Kit from Applied Biosystems™ in autosomal DNA quantification. PowerQuant ® , as compared to Quantiplex HYres and Quantifiler ® Trio, shows a better precision for both autosomal and male DNA quantifications. Quantifiler ® Trio and PowerQuant ® in contrast to Quantiplex HYres offer better correlations with lower discrepancies between autosomal and male DNA quantification, and their additional degradation index features provide a detection platform for inhibited and/or degraded DNA template. Regarding the compatibility between these quantification and profiling systems: (1) both Quantifiler ® Trio and PowerQuant ® work well with GlobalFiler and Fusion 6C, allowing a fairly accurate prediction of their DNA typing results based on the quantification values; (2) Quantiplex HYres offers a fairly reliable IPC system for detecting any potential inhibitions on Investigator 24plex, whereas Quantifiler ® Trio and PowerQuant ® suit better for Global

  17. The Influence of Environmental Friendliness on Green Trust: The Mediation Effects of Green Satisfaction and Green Perceived Quality

    Directory of Open Access Journals (Sweden)

    Yu-Shan Chen

    2015-07-01

    Full Text Available As global green trends became more prevalent, green marketing also developed into an important issue. Although prior literature explored the main factors affecting green trust, it was inconclusive as to how environmental friendliness could affect the green trust in green marketing. This study aims to focus on the positive influence of environmental friendliness on green trust, and explore the mediation effects of green satisfaction and green perceived quality. This study undertakes an empirical study by means of questionnaire survey. The respondents are consumers who have experience purchasing green products. This study applies structural equation modeling (SEM to test the hypotheses. The findings of this study indicate that (1 environmental friendliness has a significant positive impact on green satisfaction, green perceived quality, and green trust; (2 both green satisfaction and green perceived quality positively affect green trust; and (3 green satisfaction and green perceived quality partially mediate the positive relationship between environmental friendliness and green trust.

  18. Latent class analysis of real time qPCR and bacteriological culturing for the diagnosis of Streptococcus agalactiae in cow composite milk samples

    DEFF Research Database (Denmark)

    Holmøy, Ingrid H.; Toft, Nils; Jørgensen, Hannah J.

    2018-01-01

    Streptococcus agalactiae (S. agalactiae) has re-emerged as a mastitis pathogen among Norwegian dairy cows. The Norwegian cattle health services recommend that infected herds implement measures to eradicate S. agalactiae, this includes a screening of milk samples from all lactating cows....... The performance of the qPCR-test currently in use for this purpose has not been evaluated under field conditions. The objective of this study was to estimate the sensitivity and specificity of the real-time qPCR assay in use in Norway (Mastitis 4 qPCR, DNA Diagnostics A/S, Risskov, Denmark) and compare...... it to conventional bacteriological culturing for detection of S. agalactiae in milk samples. Because none of these tests are considered a perfect reference test, the evaluation was performed using latent class models in a Bayesian analysis. Aseptically collected cow-composite milk samples from 578 cows belonging...

  19. Development of a rapid HRM qPCR for the diagnosis of the four most prevalent Plasmodium lineages in New Zealand.

    Science.gov (United States)

    Schoener, E R; Hunter, S; Howe, L

    2017-07-01

    Although wildlife rehabilitation and translocations are important tools in wildlife conservation in New Zealand, disease screening of birds has not been standardized. Additionally, the results of the screening programmes are often difficult to interpret due to missing disease data in resident or translocating avian populations. Molecular methods have become the most widespread method for diagnosing avian malaria (Plasmodium spp.) infections. However, these methods can be time-consuming, expensive and are less specific in diagnosing mixed infections. Thus, this study developed a new real-time PCR (qPCR) method that was able to detect and specifically identify infections of the three most common lineages of avian malaria in New Zealand (Plasmodium (Novyella) sp. SYAT05, Plasmodium elongatum GRW6 and Plasmodium spp. LINN1) as well as a less common, pathogenic Plasmodium relictum GRW4 lineage. The assay was also able to discern combinations of these parasites in the same sample and had a detection limit of five parasites per microlitre. Due to concerns relating to the presence of the potentially highly pathogenic P. relictum GRW4 lineage in avian populations, an additional confirmatory high resolution (HRM) qPCR was developed to distinguish between commonly identified P. elongatum GRW6 from P. relictum GRW4. The new qPCR assays were tested using tissue samples containing Plasmodium schizonts from three naturally infected dead birds resulting in the identified infection of P. elongatum GRW6. Thus, these rapid qPCR assays have shown to be cost-effective and rapid screening tools for the detection of Plasmodium infection in New Zealand native birds.

  20. Quantification by qPCR of Pathobionts in Chronic Periodontitis: Development of Predictive Models of Disease Severity at Site-Specific Level

    OpenAIRE

    Tomás, Inmaculada; Regueira-Iglesias, Alba; López, Maria; Arias-Bujanda, Nora; Novoa, Lourdes; Balsa-Castro, Carlos; Tomás, Maria

    2017-01-01

    Currently, there is little evidence available on the development of predictive models for the diagnosis or prognosis of chronic periodontitis based on the qPCR quantification of subgingival pathobionts. Our objectives were to: (1) analyze and internally validate pathobiont-based models that could be used to distinguish different periodontal conditions at site-specific level within the same patient with chronic periodontitis; (2) develop nomograms derived from predictive models. Subgingival pl...

  1. Analytical Performance of Four Polymerase Chain Reaction (PCR and Real Time PCR (qPCR Assays for the Detection of Six Leishmania Species DNA in Colombia

    Directory of Open Access Journals (Sweden)

    Cielo M. León

    2017-10-01

    Full Text Available Leishmaniasis comprises a spectrum of parasitic diseases caused by protozoans of the genus Leishmania. Molecular tools have been widely employed for the detection of Leishmania due to its high sensitivity and specificity. However, the analytical performance of molecular platforms as PCR and real time PCR (qPCR including a wide variety of molecular markers has never been evaluated. Herein, the aim was to evaluate the analytical performance of 4 PCR-based assays (designed on four different targets and applied on conventional and real-time PCR platforms. We evaluated the analytical performance of conventional PCR and real time PCR, determining exclusivity and inclusivity, Anticipated Reportable Range (ARR, limit of detection (LoD and accuracy using primers directed to kDNA, HSP70, 18S and ITS-1 targets. We observed that the kDNA was the most sensitive but does not meet the criterion of exclusivity. The HSP70 presented a higher LoD in conventional PCR and qPCR in comparison with the other markers (1 × 101 and 1 × 10-1 equivalent parasites/mL respectively and had a higher coefficient of variation in qPCR. No statistically significant differences were found between the days of the test with the four molecular markers. The present study revealed that the 18S marker presented the best performance in terms of analytical sensitivity and specificity for the qPCR in the species tested (species circulating in Colombia. Therefore, we recommend to explore the analytical and diagnostic performance in future studies using a broader number of species across America.

  2. Evaluation of Suitable Reference Genes for Normalization of qPCR Gene Expression Studies in Brinjal (Solanum melongena L.) During Fruit Developmental Stages.

    Science.gov (United States)

    Kanakachari, Mogilicherla; Solanke, Amolkumar U; Prabhakaran, Narayanasamy; Ahmad, Israr; Dhandapani, Gurusamy; Jayabalan, Narayanasamy; Kumar, Polumetla Ananda

    2016-02-01

    Brinjal/eggplant/aubergine is one of the major solanaceous vegetable crops. Recent availability of genome information greatly facilitates the fundamental research on brinjal. Gene expression patterns during different stages of fruit development can provide clues towards the understanding of its biological functions. Quantitative real-time PCR (qPCR) has become one of the most widely used methods for rapid and accurate quantification of gene expression. However, its success depends on the use of a suitable reference gene for data normalization. For qPCR analysis, a single reference gene is not universally suitable for all experiments. Therefore, reference gene validation is a crucial step. Suitable reference genes for qPCR analysis of brinjal fruit development have not been investigated so far. In this study, we have selected 21 candidate reference genes from the Brinjal (Solanum melongena) Plant Gene Indices database (compbio.dfci.harvard.edu/tgi/plant.html) and studied their expression profiles by qPCR during six different fruit developmental stages (0, 5, 10, 20, 30, and 50 days post anthesis) along with leaf samples of the Pusa Purple Long (PPL) variety. To evaluate the stability of gene expression, geNorm and NormFinder analytical softwares were used. geNorm identified SAND (SAND family protein) and TBP (TATA binding protein) as the best pairs of reference genes in brinjal fruit development. The results showed that for brinjal fruit development, individual or a combination of reference genes should be selected for data normalization. NormFinder identified Expressed gene (expressed sequence) as the best single reference gene in brinjal fruit development. In this study, we have identified and validated for the first time reference genes to provide accurate transcript normalization and quantification at various fruit developmental stages of brinjal which can also be useful for gene expression studies in other Solanaceae plant species.

  3. Analytical Performance of Four Polymerase Chain Reaction (PCR) and Real Time PCR (qPCR) Assays for the Detection of Six Leishmania Species DNA in Colombia

    Science.gov (United States)

    León, Cielo M.; Muñoz, Marina; Hernández, Carolina; Ayala, Martha S.; Flórez, Carolina; Teherán, Aníbal; Cubides, Juan R.; Ramírez, Juan D.

    2017-01-01

    Leishmaniasis comprises a spectrum of parasitic diseases caused by protozoans of the genus Leishmania. Molecular tools have been widely employed for the detection of Leishmania due to its high sensitivity and specificity. However, the analytical performance of molecular platforms as PCR and real time PCR (qPCR) including a wide variety of molecular markers has never been evaluated. Herein, the aim was to evaluate the analytical performance of 4 PCR-based assays (designed on four different targets) and applied on conventional and real-time PCR platforms. We evaluated the analytical performance of conventional PCR and real time PCR, determining exclusivity and inclusivity, Anticipated Reportable Range (ARR), limit of detection (LoD) and accuracy using primers directed to kDNA, HSP70, 18S and ITS-1 targets. We observed that the kDNA was the most sensitive but does not meet the criterion of exclusivity. The HSP70 presented a higher LoD in conventional PCR and qPCR in comparison with the other markers (1 × 101 and 1 × 10-1 equivalent parasites/mL respectively) and had a higher coefficient of variation in qPCR. No statistically significant differences were found between the days of the test with the four molecular markers. The present study revealed that the 18S marker presented the best performance in terms of analytical sensitivity and specificity for the qPCR in the species tested (species circulating in Colombia). Therefore, we recommend to explore the analytical and diagnostic performance in future studies using a broader number of species across America. PMID:29046670

  4. Green technology meets ecotoxicology

    Directory of Open Access Journals (Sweden)

    Kristina Radošević

    2016-01-01

    Full Text Available By applying concept and principles of green chemistry into different technological processes, green technologies are developed. The environmental and economic benefits of “green” approach is achieved through several directions, such as the use of renewable raw materials, creation of economic efficiency, the use of alternative reaction conditions, as well as the application of non-conventional solvents. From the point view of green chemistry, alternative solvents, in order to be a “green“ substitution to hazardous organic solvents, should be: non-volatile, non-flammable, stabile, synthesized by an environmentally friendly procedure, nontoxic and biodegradable. The toxic impact of all newly synthesized chemicals, such as alternative solvents, could be determined by methods and techniques of ecotoxicology. Ecotoxicology, an interdisciplinary scientific field, can serve as a way of monitoring the greenness of the processes. In vivo and in vitro experiments are used to study the effects of chemicals on different levels of organizations, from molecules to communities and ecosystem. The usage of in vitro methods is encouraged by a scientific community and regulatory agencies as an alternative to in vivo studies in order to reduce the number of laboratory animals used in the toxicological studies. Therefore, in this paper we gave a brief overview on the usage of animal cell cultures within the field of green chemistry and technology.

  5. Labelling it green

    Energy Technology Data Exchange (ETDEWEB)

    Evans, S.; Brocklehurst, F. [ETSU, Didcot (United Kingdom)

    1998-12-31

    The first two rounds of contracts awarded through the NFFO will expire in December 1998. These generators will then be looking for new contracts to supply renewable electricity. Since these projects were initiated the renewable energy market has grown steadily, but it is still mainly restricted to the protected market within NFFO. Consumer interest has grown steadily too, fuelled by the emergence of green energy supply companies. Market research has indicated that consumers would like the choice of green electricity, what remains unclear is if they would exercise this choice and to what extent they might pay a premium price for the privilege. From September 1998 the phased introduction of domestic sector franchise de-regulation commences. In principle, consumers can purchase their electricity from any supplier. This provides a golden opportunity for green generation. To make the most of this opportunity generators and suppliers will need to clearly explain to the public what their product is, how it is different and how everyone benefits from its use. A major marketing issue will be to provide assurance to the general public, that for example, they can indeed purchase energy from a windfarm in Wales, despite living in areas other than Wales. The DTI is assisting the expansion of the green market into the domestic sector via funding a project which plans to deliver an accreditation scheme in September 1998. This will provide a means of verifying the green claims of generators/supply companies. (Author)

  6. Use of real-time qPCR to quantify members of the unculturable heterotrophic bacterial community in a deep sea marine sponge, Vetulina sp.

    Science.gov (United States)

    Cassler, M; Peterson, C L; Ledger, A; Pomponi, S A; Wright, A E; Winegar, R; McCarthy, P J; Lopez, J V

    2008-04-01

    In this report, real-time quantitative PCR (TaqMan qPCR) of the small subunit (SSU) 16S-like rRNA molecule, a universal phylogenetic marker, was used to quantify the relative abundance of individual bacterial members of a diverse, yet mostly unculturable, microbial community from a marine sponge. Molecular phylogenetic analyses of bacterial communities derived from Caribbean Lithistid sponges have shown a wide diversity of microbes that included at least six major subdivisions; however, very little overlap was observed between the culturable and unculturable microbial communities. Based on sequence data of three culture-independent Lithistid-derived representative bacteria, we designed probe/primer sets for TaqMan qPCR to quantitatively characterize selected microbial residents in a Lithistid sponge, Vetulina, metagenome. TaqMan assays included specificity testing, DNA limit of detection analysis, and quantification of specific microbial rRNA sequences such as Nitrospira-like microbes and Actinobacteria up to 172 million copies per microgram per Lithistid sponge metagenome. By contrast, qPCR amplification with probes designed for common previously cultured sponge-associated bacteria in the genera Rheinheimera and Marinomonas and a representative of the CFB group resulted in only minimal detection of the Rheiheimera in total DNA extracted from the sponge. These data verify that a large portion of the microbial community within Lithistid sponges may consist of currently unculturable microorganisms.

  7. Trading green electricity

    International Nuclear Information System (INIS)

    Davies, M.

    1997-01-01

    A study has been carried out into the feasibility of developing an electricity trading mechanism which would allow consumers to purchase electricity which has been derived from renewable energy resources. This study was part funded by the European Commission (ALTENER), the Department of Trade and Industry and a number of private sector companies. The trading mechanism is known as the Green Pool. As a result of the findings of this study discussions are being held with potential generators and suppliers to establish a Green Pool plc. The aim is to encourage the development of new renewable energy projects outside the NFFO and SRO schemes. The Green Pool plc will be owned by the generators and its main objective will be to market the electricity produced by its members. (Author)

  8. Certified: green power

    International Nuclear Information System (INIS)

    Rhodes, S.; Brown, L.

    1999-01-01

    Deregulation of the energy industry in the USA may be a force favouring the environment but for the consumer it is something of a nightmare since there are so many options with respect to both price, service and environmental awareness. However, there is now a marked tendency for companies wishing to be seen as 'green' to favour environmentally aware suppliers. Indeed, some suppliers holding formal qualifications in 'greenness' believe they are justified in charging a premium for their energy. The question is asked 'what is green?' and the authors discuss the answers at some length: the hydro industry fares well in such a discussion. The authors (from Scientific Certification Systems) believe that certification provides a rational explanation of prices and why charging a premium may be justifiable.(UK)

  9. GREEN BANKING IN ROMANIA

    Directory of Open Access Journals (Sweden)

    Dumitrascu Mihaela

    2014-07-01

    Full Text Available The paper investigates the green banking in Romania, a new approach of conducting the banking business through considering the corporate social responsibility and environmental aspects. Nowadays, it is difficult to face the globalization and competition in order to asssure the implementation of the green banking practices. The aim of the present study is to identify corporations that have sustainability concerns. To achieve this objective, we set some hypothesis and after this we showed that the corporations are more likely to be included in the list of top banks in the world. Our study is relevant for future research in this area, because of the importance of such aspects in corporations nowadays.The conclusions of our study is that green banking practices in Romania is in an incipient stage

  10. Green Logistics Management

    Science.gov (United States)

    Chang, Yoon S.; Oh, Chang H.

    Nowadays, environmental management becomes a critical business consideration for companies to survive from many regulations and tough business requirements. Most of world-leading companies are now aware that environment friendly technology and management are critical to the sustainable growth of the company. The environment market has seen continuous growth marking 532B in 2000, and 590B in 2004. This growth rate is expected to grow to 700B in 2010. It is not hard to see the environment-friendly efforts in almost all aspects of business operations. Such trends can be easily found in logistics area. Green logistics aims to make environmental friendly decisions throughout a product lifecycle. Therefore for the success of green logistics, it is critical to have real time tracking capability on the product throughout the product lifecycle and smart solution service architecture. In this chapter, we introduce an RFID based green logistics solution and service.

  11. Why Green Taxation

    DEFF Research Database (Denmark)

    Hjøllund, Lene; Svendsen, Gert Tinggaard

    2001-01-01

    According to economists solving environmental problems is simple. Politicians should simply impose a uniform tax on harmful emissions. However, the actual design of such green taxation shows that politicians do not follow their advice. CO2 taxation in OECD, for example, is highly differentiated...... and much in favour of industry. In fact, CO2 tax rates for industry are, on average, six times lower than those for households. We argue that the reason for this tax differentiation is that industry, in contrast to households, has a strong capability to lobby. Therefore, green taxation is effectively...... blocked and the desired environmental results are not being achieved. Why then is green taxation persistently applied in relation to industry? We argue that strong fiscal incentives drive this policy choice at the expense of environmental concerns because it allows environmental bureaucracies to budget-maximize....

  12. Green Chemistry Pedagogy

    Science.gov (United States)

    Kolopajlo, Larry

    2017-02-01

    This chapter attempts to show how the practice of chemistry teaching and learning is enriched by the incorporation of green chemistry (GC) into lectures and labs. To support this viewpoint, evidence from a wide range of published papers serve as a cogent argument that GC attracts and engages both science and nonscience students, enhances chemistry content knowledge, and improves the image of the field, while preparing the world for a sustainable future. Published pedagogy associated with green and sustainable chemistry is critically reviewed and discussed.

  13. GREEN CORE HOUSE

    Directory of Open Access Journals (Sweden)

    NECULAI Oana

    2017-05-01

    Full Text Available The Green Core House is a construction concept with low environmental impact, having as main central element a greenhouse. The greenhouse has the innovative role to use the biomass energy provided by plants to save energy. Although it is the central piece, the greenhouse is not the most innovative part of the Green Core House, but the whole building ensemble because it integrates many other sustainable systems as "waste purification systems", "transparent photovoltaic panels" or "double skin façades".

  14. Green in software engineering

    CERN Document Server

    Calero Munoz, Coral

    2015-01-01

    This is the first book that presents a comprehensive overview of sustainability aspects in software engineering. Its format follows the structure of the SWEBOK and covers the key areas involved in the incorporation of green aspects in software engineering, encompassing topics from requirement elicitation to quality assurance and maintenance, while also considering professional practices and economic aspects. The book consists of thirteen chapters, which are structured in five parts. First the "Introduction" gives an overview of the primary general concepts related to Green IT, discussing wha

  15. Green Chemistry Metrics with Special Reference to Green Analytical Chemistry

    OpenAIRE

    Marek Tobiszewski; Mariusz Marć; Agnieszka Gałuszka; Jacek Namieśnik

    2015-01-01

    The concept of green chemistry is widely recognized in chemical laboratories. To properly measure an environmental impact of chemical processes, dedicated assessment tools are required. This paper summarizes the current state of knowledge in the field of development of green chemistry and green analytical chemistry metrics. The diverse methods used for evaluation of the greenness of organic synthesis, such as eco-footprint, E-Factor, EATOS, and Eco-Scale are described. Both the well-establis...

  16. Sustainable house construction and green financing. Explanation for 'green mortgages'

    International Nuclear Information System (INIS)

    1997-05-01

    The Dutch government finances the sustainable construction of new houses by means of so-called 'green loans'. Extra costs for the construction of a sustainable house are compensated by a lower interest rate for a green loan. In this brochure it is explained when green financing of house construction is possible and how to apply for such loans

  17. Plants as green phones

    NARCIS (Netherlands)

    Soler, R.; Harvey, J.A.; Bezemer, T.M.; Stuefer, J.F.

    2008-01-01

    Plants can act as vertical communication channels or `green phones¿ linking soil-dwelling insects and insects in the aboveground ecosystem. When root-feeding insects attack a plant, the direct defense system of the shoot is activated, leading to an accumulation of phytotoxins in the leaves. The

  18. Putting on the green

    Science.gov (United States)

    The green chemistry movement is scrutinized for marks of tangible success in this short perspective. Beginning with the easily identified harm of the Union Carbide Bhopal, India disaster and the concerns of Love Canal site in Niagara Falls, NY the public can begin to more easily ...

  19. Green, Brown, and probability

    CERN Document Server

    Chung, Kai Lai

    1995-01-01

    This volume shows modern probabilistic methods in action: Brownian Motion Process as applied to the electrical phenomena investigated by Green et al., beginning with the Newton-Coulomb potential and ending with solutions by first and last exits of Brownian paths from conductors.

  20. Why Green Taxation

    DEFF Research Database (Denmark)

    Hjøllund, Lene; Svendsen, Gert Tinggaard

    2001-01-01

    According to economists solving environmental problems is simple. Politicians should simply impose a uniform tax on harmful emissions. However, the actual design of such green taxation shows that politicians do not follow their advice. CO2 taxation in OECD, for example, is highly differentiated...

  1. News: Green Chemistry & Technology

    Science.gov (United States)

    A series of 21 articles focused on different features of green chemistry in a recent issue of Chemical Reviews. Topics extended over a wide range to include the design of sustainable synthetic processes to biocatalysis. A selection of perspectives follows as part of this colu

  2. The Green Obligation

    Science.gov (United States)

    Adams, Cameron

    2007-01-01

    As the green movement grows, studies provide conclusive evidence about the benefits of environmentally conscious practices indoors and outdoors. Schools are no exception. Many of these studies demonstrate how poor indoor air quality (IAQ) in schools adversely affects many of the nation's 55 million students with health problems such as asthma and…

  3. A green chemistry approach

    Indian Academy of Sciences (India)

    Administrator

    One-pot synthesis of quinaldine derivatives by using microwave irradiation without any solvent – A green chemistry approach. JAVAD SAFARI*, SAYED HOSSEIN BANITABA and SEPEHR SADEGH SAMIEI. Department of Chemistry, The Faculty of sciences, University of Kashan, Kashan,. P.O. Box 87317-51167, I.R. Iran.

  4. Between green and grey

    NARCIS (Netherlands)

    Jeanet Kullberg

    2016-01-01

    Original title: Tussen groen en grijs Taking cuttings is cool. Growing vegetables is all the rage. Green oases can now be found scattered throughout Dutch towns and cities: community gardens and roof gardens where residents can go to relax and enjoy themselves, improve the appearance of their

  5. Developing Green Line Products

    DEFF Research Database (Denmark)

    Muñoz-Marin, Ana Maria; Lassen, Astrid Heidemann; Poulsen, Søren Bolvig

    2011-01-01

    This publication is based on the Master thesis “User-driven ecoinnovation process: Towards the implementation of the Green product line at JELD-WEN” written by Ana Maria Muñoz-Marin as her Graduation Project for the MSc. Global Innovation Management degree. The company-based experiment was carried...

  6. Being 'green' helps profitability?

    International Nuclear Information System (INIS)

    Austin, D.

    1999-01-01

    Pollution reduction beyond regulatory compliance is gaining momentum among firms, but managers ask if being 'green' helps profitability. Evidence suggests it doesn't hurt, but when we see environmentally attractive firms with sound financial performance, it cannot yet say which is cause and which is effect [it

  7. Lean Green Machines

    Science.gov (United States)

    Villano, Matt

    2011-01-01

    Colleges and universities have been among the leaders nationwide in adopting green initiatives, partly due to their demographics, but also because they are facing their own budget pressures. Virtualization has become the poster child of many schools' efforts, because it provides significant bang for the buck. However, more and more higher…

  8. Greening China Naturally

    Science.gov (United States)

    Shixiong Cao; Ge Sun; Zhiqiang Zhang; Liding Che; Qi Feng; et. al.

    2011-01-01

    China leads the world in afforestation, and is one of the few countries whose forested area is increasing. However, this massive “greening” effort has been less effective than expected; afforestation has sometimes produced unintended environmental, ecological, and socioeconomic consequences, and has failed to achieve the desired ecological benefits. Where afforestation...

  9. Nanoreactors for green catalysis

    NARCIS (Netherlands)

    de Martino, M.T.; Abdelmohsen, L.K.E.A.; Rutjes, Floris P.J.T.; van Hest, J.C.M.; Hessel, V.

    2018-01-01

    Sustainable and environmentally benign production are key drivers for developments in the chemical industrial sector, as protecting our planet has become a significant element that should be considered for every industrial breakthrough or technological advancement. As a result, the concept of green

  10. Green Maritime Logistics

    DEFF Research Database (Denmark)

    Psaraftis, Harilaos N.

    2016-01-01

    By green maritime logistics we mean achieving an acceptable environmental performance of the maritime transport logistical supply chain while at the same time respecting traditional economic criteria. In this paper the environmental focus is on maritime emissions. Achieving such goal may involve ...

  11. Green Software Products

    NARCIS (Netherlands)

    Jagroep, E.A.

    2017-01-01

    The rising energy consumption of the ICT industry has triggered a quest for more green, energy efficient ICT solutions. The role of software as the true consumer of power and its potential contribution to reach sustainability goals has increasingly been acknowledged. At the same time, it is shown to

  12. Frobenius Green functors

    OpenAIRE

    Baker, Andrew

    2012-01-01

    These notes provide an informal introduction to a type of Mackey functor that arises naturally in algebraic topology in connection with Morava $K$-theory of classifying spaces of finite groups. The main aim is to identify key algebraic aspects of the Green functor structure obtained by applying a Morava $K$-theory to such classifying spaces.

  13. Global Green Growth Institute

    DEFF Research Database (Denmark)

    Müller, Anders Riel

    2017-01-01

    Har man fulgt historien om Venstres gruppeformand Lars Løkkes rejser på 1. klasse i forbindelse med formandsposten for Global Green Growth Institute (GGGI) er der sikkert nogle der undrer sig over, hvad GGGI er for en størrelse. Medierne præsenterer GGGI som en international klimaorganisation, der...

  14. Green Building Tools for Tribes

    Science.gov (United States)

    Tribal green building tools and funding information to support tribal building code adoption, healthy building, siting, energy efficiency, renewable energy, water conservation, green building materials, recycling and adaptation and resilience.

  15. Green Manufacturing Fundamentals and Applications

    CERN Document Server

    2013-01-01

    Green Manufacturing: Fundamentals and Applications introduces the basic definitions and issues surrounding green manufacturing at the process, machine and system (including supply chain) levels. It also shows, by way of several examples from different industry sectors, the potential for substantial improvement and the paths to achieve the improvement. Additionally, this book discusses regulatory and government motivations for green manufacturing and outlines the path for making manufacturing more green as well as making production more sustainable. This book also: • Discusses new engineering approaches for manufacturing and provides a path from traditional manufacturing to green manufacturing • Addresses regulatory and economic issues surrounding green manufacturing • Details new supply chains that need to be in place before going green • Includes state-of-the-art case studies in the areas of automotive, semiconductor and medical areas as well as in the supply chain and packaging areas Green Manufactu...

  16. Benefits of Green Power Partnership

    Science.gov (United States)

    The U.S. EPA's Green Power Partnership is a voluntary partnership program designed to reduce the environmental impact of electricity generation by promoting renewable energy. Learn about the benefits of becoming a Green Power Partner.

  17. The green highway forum

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2006-07-01

    In late 2004, as part of American Coal Ash Association's (ACAA) strategic planning process, a plan was approved by its Board of Directors implementing a 'green highways' concept which emphasized use of coal combustion products (CCPs) in highways in a variety of ways including being used alone, in combination with other forms of CCPs, and combined with non ash materials. The incentives behind the developed concept were the derived advantages from beneficial technical economic and environmental impacts. Although the primary use of fly ash is concrete, other forms of CCPs could be considered for more non-traditional highway applications. For example, these might include soils stabilization, binders for in-place pavement recycling, use in flowable fills, aggregates, source materials for structural fills and embankments, components in manufactured soils, and for granular base courses beneath pavements. At this same time, unknown to ACCA, EPA Region 3 in Philadelphia was working with the Wetlands and Watershed Work Group, a non-profit organization involved in wetlands policy and management along with the Federal Highway Administration (FHWA) on their own Green Highways initiative. These groups were planning a conference, the 'Green Highway Forum'. This was held in College Park, Maryland at the University of Maryland, Nov 8-10 2005. At the conference a draft 'roadmap' was presented as a guide to executive level participants bringing the diverse viewpoints of many agencies and interest groups together. Ten guiding principals were considered. The 'Green Highways' is a new effort to recognize the 'greenness' of many projects already completed and those to be initiated. 2 photos.

  18. Green function on product networks

    OpenAIRE

    Arauz Lombardía, Cristina; Carmona Mejías, Ángeles; Encinas Bachiller, Andrés Marcos

    2012-01-01

    Our objective is to determine the Green function of product networks in terms of the Green function of one of the factor networks and the eigenvalues and eigenfunctions of the Schr odinger operator of the other factor network, which we consider that are known. Moreover, we use these results to obtain the Green function of spider networks in terms of Green functions over cicles and paths. Peer Reviewed

  19. GREEN PACKAGING, GREEN PRODUCT, GREEN ADVERTISING, PERSEPSI, DAN MINAT BELI KONSUMEN

    Directory of Open Access Journals (Sweden)

    Imam Santoso

    2016-12-01

    Full Text Available Environmental problems become one of the strategic issues in achieving global competitiveness. One of the issues is products that are made from environmental friendly materials or known as green product. Furthermore, in green products marketing, the company also uses green packaging and green advertising concept. This study aimed to analyze the effect of green packaging, green products, and green advertising on consumer perception and purchasing intention. The study was conducted in Ketawanggede Village, Lowokwaru Sub-district, Malang City. The sampling method used nonprobability accidential sampling techniques. The numbers of respondents were 113 consumers in study site. Data were collected by interview using questionnaires. The method of analysis used Generalized Structured Component Analysis (GSCA. The analysis showed that the green packaging, green products, and green advertising had positive significant influence on consumer perceptions. Meanwhile, green product and consumer perception had positive significant influence on purchasing interest, but the green packaging and green advertising has not found sufficient evidence in influencing purchasing intention.

  20. It's Not Easy Building Green.

    Science.gov (United States)

    Higgins, Joseph

    2003-01-01

    Discusses green buildings, facilities designed, constructed, and operated in an environmentally friendly and resource-efficient way. Discusses reasons for campuses to "go green," the "shades of green" or variations in environmental-friendliness, certification through the Leadership in Energy and Environmental Design (LEED) rating system, financial…

  1. Green mortgages in the Netherlands

    International Nuclear Information System (INIS)

    Bosch, N.

    1997-01-01

    Since November 1996 sustainable building of houses is also part of the fiscal Regulation for Green Projects (i.e. the stimulation of environment-friendly investments). The extension of that financial regulation resulted in a new product: Green Mortgages. The conditions that have to be met to be qualified for a Green Mortgage are briefly outlined

  2. Green economy and related concepts

    NARCIS (Netherlands)

    Loiseau, Eleonore; Saikku, Laura; Antikainen, Riina; Droste, Nils; Hansjürgens, Bernd; Pitkänen, Kati; Leskinen, Pekka; Kuikman, Peter; Thomsen, Marianne

    2016-01-01

    For the last ten years, the notion of a green economy has become increasingly attractive to policy makers. However, green economy covers a lot of diverse concepts and its links with sustainability are not always clear. In this article, we focus on definitions of green economy and related concepts

  3. Guide to Purchasing Green Power

    Science.gov (United States)

    The Guide for Purchasing Green Power is a comprehensive guide for current and potential buyers of green power with information about green power purchasing. The Guide is created cooperatively between the EPA, the U.S. Department of Energy, the World Resou

  4. Investing in a green future

    Science.gov (United States)

    Clapp, Christa

    2018-01-01

    The growing green bond market reflects the financial sector's awakening to climate risk. New research examining the US municipal bond market suggests a positive green bond premium in recent years, driven by differences in credit quality. As climate-risk disclosure becomes more widespread, investors may show willingness to pay green premiums.

  5. Green Roofs for Stormwater Management

    Science.gov (United States)

    This project evaluated green roofs as a stormwater management tool. Results indicate that the green roofs are capable of removing 40% of the annual rainfall volume from a roof through retention and evapotranspiration. Rainfall not retained by green roofs is detained, effectively...

  6. The use of kDNA minicircle subclass relative abundance to differentiate between Leishmania (L.) infantum and Leishmania (L.) amazonensis.

    Science.gov (United States)

    Ceccarelli, Marcello; Galluzzi, Luca; Diotallevi, Aurora; Andreoni, Francesca; Fowler, Hailie; Petersen, Christine; Vitale, Fabrizio; Magnani, Mauro

    2017-05-16

    Leishmaniasis is a neglected disease caused by many Leishmania species, belonging to subgenera Leishmania (Leishmania) and Leishmania (Viannia). Several qPCR-based molecular diagnostic approaches have been reported for detection and quantification of Leishmania species. Many of these approaches use the kinetoplast DNA (kDNA) minicircles as the target sequence. These assays had potential cross-species amplification, due to sequence similarity between Leishmania species. Previous works demonstrated discrimination between L. (Leishmania) and L. (Viannia) by SYBR green-based qPCR assays designed on kDNA, followed by melting or high-resolution melt (HRM) analysis. Importantly, these approaches cannot fully distinguish L. (L.) infantum from L. (L.) amazonensis, which can coexist in the same geographical area. DNA from 18 strains/isolates of L. (L.) infantum, L. (L.) amazonensis, L. (V.) braziliensis, L. (V.) panamensis, L. (V.) guyanensis, and 62 clinical samples from L. (L.) infantum-infected dogs were amplified by a previously developed qPCR (qPCR-ML) and subjected to HRM analysis; selected PCR products were sequenced using an ABI PRISM 310 Genetic Analyzer. Based on the obtained sequences, a new SYBR-green qPCR assay (qPCR-ama) intended to amplify a minicircle subclass more abundant in L. (L.) amazonensis was designed. The qPCR-ML followed by HRM analysis did not allow discrimination between L. (L.) amazonensis and L. (L.) infantum in 53.4% of cases. Hence, the novel SYBR green-based qPCR (qPCR-ama) has been tested. This assay achieved a detection limit of 0.1 pg of parasite DNA in samples spiked with host DNA and did not show cross amplification with Trypanosoma cruzi or host DNA. Although the qPCR-ama also amplified L. (L.) infantum strains, the C q values were dramatically increased compared to qPCR-ML. Therefore, the combined analysis of C q values from qPCR-ML and qPCR-ama allowed to distinguish L. (L.) infantum and L. (L.) amazonensis in 100% of tested samples

  7. Green Consumerism : an Eco-Friendly Behaviour Form Through The Green Product Consumption and Green Marketing

    OpenAIRE

    Handayani, Wiwik

    2017-01-01

    This research is referred to analyse the influence of consumer attitude of green product towards purchase intention. The consumer attitude of green product is a psychological tendencies that is expressed by evaluating a certain entity with some advantage or disadvantage considerations. The problem of this research is the low of cunsumer awareness to consume green product, because the lack to comprehend the importance of green product usage for health and eco-friendly. The purpose of this rese...

  8. Interlaboratory comparison of three microbial source tracking quantitative polymerase chain reaction (qPCR) assays from fecal-source and environmental samples

    Science.gov (United States)

    Stelzer, Erin A.; Strickler, Kriston M.; Schill, William B.

    2012-01-01

    During summer and early fall 2010, 15 river samples and 6 fecal-source samples were collected in West Virginia. These samples were analyzed by three laboratories for three microbial source tracking (MST) markers: AllBac, a general fecal indicator; BacHum, a human-associated fecal indicator; and BoBac, a ruminant-associated fecal indicator. MST markers were analyzed by means of the quantitative polymerase chain reaction (qPCR) method. The aim was to assess interlaboratory precision when the three laboratories used the same MST marker and shared deoxyribonucleic acid (DNA) extracts of the samples, but different equipment, reagents, and analyst experience levels. The term assay refers to both the markers and the procedure differences listed above. Interlaboratory precision was best for all three MST assays when using the geometric mean absolute relative percent difference (ARPD) and Friedman's statistical test as a measure of interlaboratory precision. Adjustment factors (one for each MST assay) were calculated using results from fecal-source samples analyzed by all three laboratories and applied retrospectively to sample concentrations to account for differences in qPCR results among labs using different standards and procedures. Following the application of adjustment factors to qPCR results, ARPDs were lower; however, statistically significant differences between labs were still observed for the BacHum and BoBac assays. This was a small study and two of the MST assays had 52 percent of samples with concentrations at or below the limit of accurate quantification; hence, more testing could be done to determine if the adjustment factors would work better if the majority of sample concentrations were above the quantification limit.

  9. Effectiveness of onsite wastewater reuse system in reducing bacterial contaminants measured with human-specific IMS/ATP and qPCR.

    Science.gov (United States)

    Agidi, Senyo; Vedachalam, Sridhar; Mancl, Karen; Lee, Jiyoung

    2013-01-30

    Water shortages and the drive to recycle is increasing interest in reuse of reclaimed wastewater. Timely and cost-effective ways to detect fecal pollutants prior to reuse increases confidence of residents and neighbors concerned about reuse of reclaimed wastewater. The on-site wastewater treatment and reuse systems (OWTRS) used in this study include a septic tank, peat bioreactor, ClO(2) disinfection and land spray irrigation system. Bacteroides fragilis, Escherichia coli and Enterococcus spp., were tested with immunomagnetic separation/ATP bioluminescence (IMS/ATP), qPCR and culture-based methods. The results displayed a 2-log reduction in fecal bacteria in the peat bioreactor and a 5-log reduction following chloride dioxide disinfection. The fecal bacteria levels measured by IMS/ATP correlated with qPCR results: HuBac 16S (R(2) = 0.903), Bf-group 16S (R(2) = 0.956), gyrB (R(2) = 0.673), and Ent 23S (R(2) = 0.724). This is the first study in which the newly developed human-specific IMS/ATP and previously developed IMS/ATP were applied for determining OWTRS efficiency. Results of the study revealed that IMS/ATP is a timely and cost-effective way to detect fecal contaminants, and results were validated with qPCR and culture based methods. The new IMS/ATP can also be applied broadly in the detection of human-originated fecal contamination. Copyright © 2012 Elsevier Ltd. All rights reserved.

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

    Science.gov (United States)

    Ikten, Cengiz; Ustun, Rustem; Catal, Mursel; Yol, Engin; Uzun, Bulent

    2016-01-01

    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.

  11. 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.

  12. Fluorescence in situ hybridization and qPCR to detect Merkel cell polyomavirus physical status and load in Merkel cell carcinomas.

    Science.gov (United States)

    Haugg, Anke M; Rennspiess, Dorit; zur Hausen, Axel; Speel, Ernst-Jan M; Cathomas, Gieri; Becker, Jürgen C; Schrama, David

    2014-12-15

    The Merkel cell polyomavirus (MCPyV) is detected in 80% of Merkel cell carcinomas (MCC). Clonal integration and tumor-specific mutations in the large T antigen are strong arguments that MCPyV is a human tumor virus. However, the relationship between viral presence and cancer induction remains discussed controversially. Since almost all studies on virus prevalence are based on PCR techniques, we performed MCPyV fluorescence in situ hybridization (FISH) on MCC to gain information about the quality of the viral presence on the single cell level. MCPyV-FISH was performed on tissue microarrays containing 62 formalin-fixed and paraffin-embedded tissue samples including all tumor grades of 42 patients. The hybridization patterns were correlated to the qPCR data determined on corresponding whole tissue sections. Indeed, MCPyV-FISH and qPCR data were highly correlated, i.e. 83% for FISH-positive and 93% for FISH-negative cores. Accordingly, the mean of the qPCR values of all MCPyV-positive cores differed significantly from the mean of the negative cores (p = 0.0076). Importantly, two hybridization patterns were definable in the MCPyV-FISH: a punctate pattern (85%) indicating viral integration, which correlated with a moderate viral abundance and a combination of the punctate with a diffuse pattern (15%), suggesting a possible coexistence of integrated and episomal virus which was associated with very high viral load and VP1 expression. Thus, MCPyV-FISH adds important information on the single cell level within the histomorphological context and could therefore be an important tool to further elucidate MCPyV related carcinogenesis. © 2014 UICC.

  13. Babesia bovis and Babesia bigemina infection levels estimated by qPCR in Angus cattle from an endemic area of São Paulo state, Brazil.

    Science.gov (United States)

    Giglioti, R; Oliveira, H N; Santana, C H; Ibelli, A M G; Néo, T A; Bilhassi, T B; Rabelo, M D; Machado, R Z; Brito, L G; Oliveira, M C S

    2016-07-01

    The levels of infection by Babesia bovis and Babesia bigemina were estimated by absolute quantification through the quantitative PCR technique (qPCR). Fifty-one contemporaneous Angus cattle were evaluated on two occasions. The number of standard female Rhipicephalus microplus ticks present on the left side of the body was counted and blood samples were drawn from the tail vein into tubes containing the anticoagulant EDTA. The blood samples were submitted to DNA extraction and used to quantify the number of copies (NC) of DNA from B. bovis and B. bigemina by qPCR. The data on tick count and number of DNA copies were transformed for normalization and analyzed by a mixed model method. A multivariate model with repeated measures of the same animal, including the effects of collection, parasite species and their interaction, was used. The repeatability values were obtained from the matrix of (co)variances and were expressed for each species. The correlations between the counts of different species on the same animal, in the same collection or different collections, were also estimated. The results showed the qPCR could distinguish the two between infection by the two Babesia species. Infection levels by B. bovis and B. bigemina were detected in 100% and 98% of the animals, respectively. Significant differences were found (Phemoparasites did not depend on the tick infestation levels at the moment of each collection. The repeatability values estimated indicate that under the study conditions, the variations in the tick infestation levels and of parasitemia by B. bovis and B. bigemina depend more on factors related to each collection than on intrinsic factors of the animal. Copyright © 2016 Elsevier GmbH. All rights reserved.

  14. Green roofs; Les toitures vegetalisees

    Energy Technology Data Exchange (ETDEWEB)

    Seghier, C.

    2006-03-15

    Impervious surface coverage keeps spreading in cities. Streets, sidewalks, parking lots and roofs are waterproof, meaning greater amounts of water to channel and treat and higher flood risks during heavy rainfalls. Green roofing can play a key part in addressing this alarming issue. There are three types of green roofs: extensive, semi-intensive and intensive. The extensive green roof technique uses a thin soil covering with a variety of species providing year-round plant coverage. The plants are not necessarily horticultural in which case routine maintenance is minimal. No watering is needed. Usually extensive green roofs create an ecosystem. The semi-intensive green roof technique uses a soil covering of average thickness and serves to create decorative roofing. Although maintenance is moderate, watering is essential. The intensive green roof technique produces a terrace roof garden. Another advantage of green roofs is they increase the life cycle of the sealing. Roof sealing protection may see the span of its life cycle, now at about fifteen years, doubled if the building has a green roof. planning professionals still know very little about green roofing solutions. Yet, green roofing provides unquestionable ecological qualities and thermal and acoustic performance that have proven to be environmentally friendly. Yet France lags behind northern European countries in green roofing. The Germans, Swiss, Austrians, Scandinavians and Dutch have been using the technique for more than twenty years. (A.L.B.)

  15. Green buildings pay

    DEFF Research Database (Denmark)

    Naboni, Emanuele; Edwards, Brian

    The new edition of ‘Green Buildings Pay’ authored by Brian Edwards and Emanuele Naboni explores the business and professional benefits which derive from architectural design driven by sustainability. With a new sub-title ‘Green Buildings Pay: design, productivity and ecology’ the book argues...... that environmental design has altered how we design, construct and manage buildings. The book has relevance to those who not only design and engineer buildings but to those who commission architecture and those who occupy the products of this process. Hence, the user is a key consideration. The book examines via...... a number of LEED and BREEAM cases the buildings which flow from corporate environmental responsibility. A number of office and university buildings are examined from three main perspectives- the architect, client and user. One key finding is that architectural innovation has been driven by ecological...

  16. The green protest

    International Nuclear Information System (INIS)

    Brun, R.

    1978-01-01

    The increasing public interest in the 'green' and the 'coloured' made the politicians alert. The ecological movement and the public initiatives show their effects. What 'utopists', 'crazy people' think about large technical development gains public importance. The uneasiness about a social development which was directed only by increasing spending power and permanent expansion, begins to articulate itself. Values such as solidarity and community, considerateness for fellow men and environment which had been eliminated in the past, are regaining their meaning. From the point of view of this steadily increasing movement, the phenomenon of the 'green parties' is investigated. In this context, the persons involved give their opinions, as well as representatives of the established parties, thus giving this volume the character of a documentation on an important trend in today's events. (orig.) [de

  17. Green and lean management

    CERN Document Server

    Davim, J

    2017-01-01

    This book focusses on the challenges and changes organizational management faces in an era when the need to develop environmentally aware processes meets high levels of competition. It covers the synergetic effects, how re-use, recycling, waste reduction, and other sustainable production strategies can add value, low costs and time of production. Sustainable business behavior is not only an environmental perspective on management, but more and more contains an organizational perspective. Taking into account these issues, green and lean management appears as the way managers can drive their employees to continuously improve the management processes that add value to the organization and costumers. This book provides information on principles, strategies, models, and applications of green and lean management, and at the same time communicates the latest research activity relating to this scientific field world-wide.

  18. Towards green construction

    International Nuclear Information System (INIS)

    Bajracharya, Bijaya B.; Shrestha, Prasanna M.

    2000-01-01

    Sustainability is the key to any development works. In the operation phase, hydro power is the most sustainable form of energy. However construction activities for the same power station are usually far from being green. The popular myth is that construction activity converts green into grey. Despite this popular myth, construction of a hydro power project in Nepal has made the project area greener than earlier during the construction phase itself. Choice of construction technology, appropriate level of environmental impact assessment, monitoring of environmental parameters along side the construction progress followed by mitigation at the right time; launching community development programmes side by side, having environmental specification in contractual documents and self-reliance to fulfill environmental obligations by contractors itself are the key factors in the environmental management within the construction activities. The main conclusions in the paper is the need to change the way to think about the project constraints

  19. of Brilliant Green

    Directory of Open Access Journals (Sweden)

    Rana Seyrani

    2016-01-01

    Full Text Available Novel hydrogel nanocomposites, based on κ-carrageenan polysaccharide, were prepared by graft copolymerization of acrylamide (AAM and maleic anhydride (MAH as comonomers in the presence of multiwall carbon nanotubes (MWCNT, using methylene bisacrylamide (MBA and ammonium persulfate (APS,former as a crosslinking agent and the latter as an initiator. The hydrogel nanocomposites structure was characterized by FTIR spectroscopy, scanning electron microscopy (SEM and XRD patterns, and their thermal stability was investigated by TGA thermal analysis. The hydrogel nanocomposites were evaluated using gel content measurements and swelling rate in distilled water and in saline solutions. The carbon nanotube content was examined in relation to its effect on the properties of nanocomposites. The results showed that with increasing carbon nanotube content, the rate of water absorbency and equilibrium swelling in distilled water decreased whereas the water absorbency in the saline solutions increased. Water retention capacity was also studied and the results indicated that the inclusion of carbon nanotube increased water retention under heating condition. Furthermore, the experimental conditions of adsorption kinetics and dynamics for the removal of cationic dye, Brilliant Green (BG, were studied in the range of 6-8 for pH, 10-60 min for time (t, and 10-300 mg/L for initial concentration (C0 of the dye. The optimum conditions obtained for adsorption of Brilliant Green dye were pH 7, t= 50 min and C0= 10 mg/L. Also, the results indicated that more than 98% of the maximum adsorption capacity toward Brilliant Green dye was achieved within the initial 10 min. The experimental tests showed that the hydrogels could be used as fast–responsive and high capacity sorbents in Brilliant Green removal processes from industrial waste water.

  20. Green dentistry: The future

    Directory of Open Access Journals (Sweden)

    Mensudar Rathakrishnan

    2017-01-01

    Full Text Available Dentistry is a profession dedicated to promote health and wellness of the people. Green dentistry is a relatively new emerging concept in dentistry. Most dental offices are privately-owned small businesses and have no financial advantage to invest in many environmental-friendly practices. For this reason, more research is needed to find cost-effective environmental alternatives in dentistry. The aim of this article is to expand awareness and practice of eco-friendly dentistry.

  1. Evaluation of a low-cost procedure for sampling, long-term storage, and extraction of RNA from blood for qPCR analyses

    DEFF Research Database (Denmark)

    Mærkedahl, Rasmus Baadsgaard; Frøkiær, Hanne; Lauritzen, Lotte

    2015-01-01

    Abstract Background: In large clinical trials, where RNA cannot be extracted immediately after sampling, preserving RNA in whole blood is a crucial initial step in obtaining robust qPCR data. The current golden standard for RNA preservation is costly and designed for time-consuming column-based RNA....../binding solution over time and between samples stored and extracted by the two systems. Conclusions: The MagMAX system can be used for storage of human blood for up to 4 months and is equivalent to the PAXgene system for RNA extraction. It furthermore, provides a means for significant cost reduction in large...

  2. The Influence of Consumers Perception of Green Products on Green Purchase Intention

    OpenAIRE

    Wilson Kong; Amran Harun; Rini Suryati Sulong; Jaratin Lily

    2014-01-01

    Green consumerism has increasingly received attention since the increased level of consumer awareness towards green products. Therefore, the aim of this paper had been to examine the influence of consumer perception of green products on green purchase intention. In this study, perception of green products was conceptualized as a multidimensional variable comprised of green corporate perception, eco-label, green advertising, green packaging, and green product value. By using a survey, a total ...

  3. Green Chemistry Metrics with Special Reference to Green Analytical Chemistry

    Directory of Open Access Journals (Sweden)

    Marek Tobiszewski

    2015-06-01

    Full Text Available The concept of green chemistry is widely recognized in chemical laboratories. To properly measure an environmental impact of chemical processes, dedicated assessment tools are required. This paper summarizes the current state of knowledge in the field of development of green chemistry and green analytical chemistry metrics. The diverse methods used for evaluation of the greenness of organic synthesis, such as eco-footprint, E-Factor, EATOS, and Eco-Scale are described. Both the well-established and recently developed green analytical chemistry metrics, including NEMI labeling and analytical Eco-scale, are presented. Additionally, this paper focuses on the possibility of the use of multivariate statistics in evaluation of environmental impact of analytical procedures. All the above metrics are compared and discussed in terms of their advantages and disadvantages. The current needs and future perspectives in green chemistry metrics are also discussed.

  4. Green Chemistry Metrics with Special Reference to Green Analytical Chemistry.

    Science.gov (United States)

    Tobiszewski, Marek; Marć, Mariusz; Gałuszka, Agnieszka; Namieśnik, Jacek

    2015-06-12

    The concept of green chemistry is widely recognized in chemical laboratories. To properly measure an environmental impact of chemical processes, dedicated assessment tools are required. This paper summarizes the current state of knowledge in the field of development of green chemistry and green analytical chemistry metrics. The diverse methods used for evaluation of the greenness of organic synthesis, such as eco-footprint, E-Factor, EATOS, and Eco-Scale are described. Both the well-established and recently developed green analytical chemistry metrics, including NEMI labeling and analytical Eco-scale, are presented. Additionally, this paper focuses on the possibility of the use of multivariate statistics in evaluation of environmental impact of analytical procedures. All the above metrics are compared and discussed in terms of their advantages and disadvantages. The current needs and future perspectives in green chemistry metrics are also discussed.

  5. Reduction of malachite green to leucomalachite green by intestinal bacteria.

    OpenAIRE

    Henderson, A L; Schmitt, T C; Heinze, T M; Cerniglia, C E

    1997-01-01

    Intestinal microfloras from human, rat, mouse, and monkey fecal samples and 14 pure cultures of anaerobic bacteria representative of those found in the human gastrointestinal tract metabolized the triphenylmethane dye malachite green to leucomalachite green. The reduction of malachite green to the leuco derivative suggests that intestinal microflora could play an important role in the metabolic activation of the triphenylmethane dye to a potential carcinogen.

  6. Powerful qPCR assays for the early detection of latent invaders: interdisciplinary approaches in clinical cancer research and plant pathology.

    Science.gov (United States)

    Luchi, Nicola; Capretti, Paolo; Pazzagli, Mario; Pinzani, Pamela

    2016-06-01

    Latent invaders represent the first step of disease before symptoms occur in the host. Based on recent findings, tumors are considered to be ecosystems in which cancer cells act as invasive species that interact with the native host cell species. Analogously, in plants latent fungal pathogens coevolve within symptomless host tissues. For these reasons, similar detection approaches can be used for an early diagnosis of the invasion process in both plants and humans to prevent or reduce the spread of the disease. Molecular tools based on the evaluation of nucleic acids have been developed for the specific, rapid, and early detection of human diseases. During the last decades, these techniques to assess and quantify the proliferation of latent invaders in host cells have been transferred from the medical field to different areas of scientific research, such as plant pathology. An improvement in molecular biology protocols (especially referring to qPCR assays) specifically designed and optimized for detection in host plants is therefore advisable. This work is a cross-disciplinary review discussing the use of a methodological approach that is employed within both medical and plant sciences. It provides an overview of the principal qPCR tools for the detection of latent invaders, focusing on comparisons between clinical cancer research and plant pathology, and recent advances in the early detection of latent invaders to improve prevention and control strategies.

  7. Correlation of crAssphage-based qPCR markers with culturable and molecular indicators of human fecal pollution in an impacted urban watershed.

    Science.gov (United States)

    Stachler, Elyse; Akyon, Benay; Aquino de Carvalho, Nathalia; Ference, Christian; Bibby, Kyle

    2018-06-06

    Environmental waters are monitored for fecal pollution to protect public health. Many previously developed human-specific fecal pollution indicators lack adequate sensitivity to be reliably detected in environmental waters or do not correlate well with viral pathogens. Recently, two novel human sewage-associated source tracking qPCR markers were developed based on the bacteriophage crAssphage, CPQ_056 and CPQ_064. These assays are highly human specific, abundant in sewage, and are viral-based, suggesting great promise for environmental application as human fecal pollution indicators. A 30-day sampling study was conducted in an urban stream impacted by combined sewer overflows to evaluate the crAssphage markers' performance in an environmental system. The crAssphage markers were present at concentrations of 4.02-6.04 log10 copies/100 mL throughout the study period, indicating their high abundance and ease of detection in polluted environmental waters. In addition, the crAssphage assays were correlated with rain events, molecular markers for human polyomavirus and HF183, as well as culturable E. coli, enterococci, and somatic coliphage. The CPQ_064 assay correlated strongly to a greater number of biological indicators than the CPQ_056 assay. This study is the first to evaluate both crAssphage qPCR assays in an extended environmental application of crAssphage markers for monitoring of environmental waters. It is also the first study to compare crAssphage marker concentration with other viral-based indicators.

  8. The Development of a Novel qPCR Assay-Set for Identifying Fecal Contamination Originating from Domestic Fowls and Waterfowl in Israel

    Directory of Open Access Journals (Sweden)

    Shoshanit eOhad

    2016-02-01

    Full Text Available The emerging Microbial Source Tracking (MST methodologies aim to identify fecal contamination originating from domestic and wild animals, and from humans. Avian MST is especially challenging, primarily because the Aves class includes both domesticated and wild species with highly diverse habitats and dietary characteristics. The quest for specific fecal bacterial MST markers can be difficult with respect to attaining sufficient assay sensitivity and specificity. The present study utilizes High Throughput Sequencing (HTS to screen bacterial 16S rRNA genes from fecal samples collected from both domestic and wild avian species. Operational taxonomic unit (OTU analysis was then performed, from which sequences were retained for downstream qPCR marker development. Identification of unique avian host DNA sequences, absent in non-avian hosts, was then carried out using a dedicated database of bacterial 16S rRNA gene taken from the Ribosomal Database Project. Six qPCR assays were developed targeting the 16S rRNA gene of Lactobacillus, Gallibacterium, Firmicutes, Fusobacteriaceae and other bacteria. Two assays (Av4143 and Av163 identified most of the avian fecal samples and demonstrated sensitivity values of 91% and 70%, respectively. The Av43 assay only identified droppings from battery hens and poultry, whereas each of the other three assays (Av24, Av13, and Av216 identified waterfowl species with lower sensitivities values. The development of an MST assay-panel, which includes both domestic and wild avian species, expands the currently known MST analysis capabilities for decoding fecal contamination.

  9. Comparative validation using quantitative real-time PCR (qPCR and conventional PCR of bovine semen centrifuged in continuous density gradient

    Directory of Open Access Journals (Sweden)

    M.V. Resende

    2011-06-01

    Full Text Available The objective of the present study was to determine the sperm enrichment with X-bearing spermatozoa, after one centrifugation in a Percoll or OptiPrep continuous density gradient, using quantitative real-time polymerase chain reaction (qPCR of sperm DNA and resultant in vitro-produced bovine embryos by PCR. Frozen/thawed sperm was layered on density gradients and the tubes were centrifuged. Supernatants were gently aspirated and the sperm recovered from the bottom of the tubes. Cleavage and blastocyst rates were determined through in vitro production of embryos and PCR was performed to identify the embryos' genetic sex. A difference in blastocyst rate was found in the Percoll treatment compared to OptiPrep (P<0.05. The percentage of female embryos in the Percoll and OptiPrep groups was 62.0% and 47.1%, respectively. These results were confirmed by qPCR of spermatozoa DNA and underestimation was seen only in the Percoll group. It was possible to sexing sperm using simple approach.

  10. Isolation and Identification of Lactic Acid Bacteria from Traditional Dairy Products in Baotou and Bayannur of Midwestern Inner Mongolia and q-PCR Analysis of Predominant Species

    Science.gov (United States)

    2016-01-01

    In this study, traditional culture method and 16S rRNA gene analysis were applied to reveal the composition and diversity of lactic acid bacteria (LAB) of fermented cow milk, huruud and urum from Baotou and Bayannur of midwestern Inner Mongolia. Also, the quantitative results of dominant LAB species in three different types of dairy products from Baotou and Bayannur were gained by quantitative polymerase chain reaction (q-PCR) technology. Two hundred and two LAB strains isolated from sixty-six samples were identified and classified into four genera, namely Enterococcus, Lactococcus, Lactobacillus, Leuconostoc, and twenty-one species and subspecies. From these isolates, Lactococcus lactis subsp. lactis (32.18%), Lactobacillus plantarum (12.38%) and Leuconosto mesenteroides (11.39%) were considered as the dominated LAB species under the condition of cultivating in MRS and M17 medium. And the q-PCR results revealed that the number of dominant species varied from samples to samples and from region to region. This study clearly shows the composition and diversity of LAB existing in fermented cow milk, huruud and urum, which could be considered as valuable resources for LAB isolation and further probiotic selection. PMID:27621691

  11. Quantification of B16 Melanoma Cells in Lungs Using Triplex Q-PCR - A New Approach to Evaluate Melanoma Cell Metastasis and Tumor Control

    DEFF Research Database (Denmark)

    Sorensen, Maria R; Pedersen, Sara R; Lindkvist, Annika

    2014-01-01

    of survival once the tumor has metastasized. In the present study, we have developed a new assay for quantitative analysis of B16 melanoma metastasis in the lungs. We have used a triplex Q-PCR to determine the expression of the melanoma genes GP100/Pmel and tyrosinase-related protein 2 (TRP-2), and found...... that B16.F10gp cells were detectable in the lungs as early as 2 hours after intravenous challenge with ≥10(4) tumor cells. When investigating the gene expression as a function of time, we observed a gradual decrease from 2-24 hours post tumor challenge followed by an increase of approximately 2 log10...... the outgrowth of subcutaneous melanomas. Results obtained using Q-PCR were compared to conventional counting of metastatic foci under a dissection microscope. A marked reduction in gene expression was observed in the lungs after vaccination with both vectors; however, Ad-Ii-GP showed the highest protection...

  12. The Green Experiment: Cities, Green Stormwater Infrastructure, and Sustainability

    Directory of Open Access Journals (Sweden)

    Christopher M. Chini

    2017-01-01

    Full Text Available Green infrastructure is a unique combination of economic, social, and environmental goals and benefits that requires an adaptable framework for planning, implementing, and evaluating. In this study, we propose an experimental framework for policy, implementation, and subsequent evaluation of green stormwater infrastructure within the context of sociotechnical systems and urban experimentation. Sociotechnical systems describe the interaction of complex systems with quantitative and qualitative impacts. Urban experimentation—traditionally referencing climate change programs and their impacts—is a process of evaluating city programs as if in a laboratory setting with hypotheses and evaluated results. We combine these two concepts into a singular framework creating a policy feedback cycle (PFC for green infrastructure to evaluate municipal green infrastructure plans as an experimental process within the context of a sociotechnical system. After proposing and discussing the PFC, we utilize the tool to research and evaluate the green infrastructure programs of 27 municipalities across the United States. Results indicate that green infrastructure plans should incorporate community involvement and communication, evaluation based on project motivation, and an iterative process for knowledge production. We suggest knowledge brokers as a key resource in connecting the evaluation stage of the feedback cycle to the policy phase. We identify three important needs for green infrastructure experimentation: (i a fluid definition of green infrastructure in policy; (ii maintenance and evaluation components of a green infrastructure plan; and (iii communication of the plan to the community.

  13. Buying and selling green: deregulation and green power marketing

    International Nuclear Information System (INIS)

    Evans, Andrew

    2000-01-01

    This article discusses the increasing trend towards deregulation of electricity markets, and the driving forces for liberalisation in the EU and North America. The use of green tariffs offered by utilities to differentiate themselves from competitors and to gain and keep customers is reported, and the situation with regard to green energy within the deregulated electricity markets in Australia, the EU, Denmark, Finland, Germany, the Netherlands, Portugal, Sweden, the UK, Canada and the USA is outlined. Customers switching as a result of green tariffs, the growing role of renewables, and opportunities for the promotion of green tariffs are discussed. (UK)

  14. Green roofs and the LEED green building rating system

    Energy Technology Data Exchange (ETDEWEB)

    Kula, R. [Sustainable Solutions Inc., Wagoner, OK (United States)

    2005-07-01

    The sustainable building industry is becoming increasingly aware of the host of public and private benefits that green roofs can provide in built environments. In dense urban environments, green roofs function to reduce stormwater runoff, urban heat island effects, and particulate matter (PM) pollution. The emerging green roof industry is now poised to support the efforts of green building networks in North America. This paper discussed the general benefits of green roofs, and their recognition within the Leadership in Energy and Environmental Design (LEED) Green Building Rating System. A case study of Mountain Equipment Co-op's Winnipeg site was presented. The building's green roof was directly responsible for earning 5 credits and contributing to the achievement of an additional 2 credits under the LEEDS certification process. Credits were earned for reduced site disturbance; landscape design to reduce heat islands; and water efficiency. The green roof at the site provided the vast majority of the building's cooling needs through an evaporative cooling trough. A photovoltaic pump was used to feed the building's irrigation system, as well as to pump ground water through cooling valances. It was concluded that the rise of sustainable building practices and the LEED Green Building Rating System will revolutionize the way new buildings are constructed.

  15. The Influence of Proactive Green Innovation and Reactive Green Innovation on Green Product Development Performance: The Mediation Role of Green Creativity

    OpenAIRE

    Yu-Shan Chen; Tai-Wei Chang; Chun-Yu Lin; Pi-Yu Lai; Kuan-Hung Wang

    2016-01-01

    This study fills the research gap in the exploration of the relationships between both proactive and reactive green innovations and green product development performance, and examines the mediating effect of green creativity. Structural equation modeling (SEM) is utilized to test the hypotheses. From the sample of 146 valid respondents, the results show that proactive green innovation positively affects green creativity and green product development performance, and green creativity positivel...

  16. Commercial green energy. Final report

    International Nuclear Information System (INIS)

    Kalweit, B.

    1998-11-01

    Firms offering a Green electricity product have discovered that residential customers are willing to pay extra for the assurance that their electricity is generated through the use of non-polluting or renewable resources. This research investigated the market potential for Green energy at the next level of the energy consuming chain, commercial establishments at which small and medium sized businesses interface with customers. Green energy is proving to be an attractive proposition to some consumers in the residential marketplace. Is there a possibility that Green energy can also be sold to commercial enterprises? This research project sought to answer this question and to investigate the factors that might lead small business people to opt for Green. Answers to these questions will help energy companies target the businesses most likely to accept Green power with the right product set and product features

  17. Picturing Urban Green Attachments

    DEFF Research Database (Denmark)

    Blok, Anders; Meilvang, Marie Leth

    2015-01-01

    In this article, we explore the cultural-political tensions and ambiguities of urban ecology, by way of following how activists move and translate between ‘familiar’ and ‘public’ engagements in the green city. Empirically, we locate our exploration in and around Nordhavnen (The North Harbor......), a large-scale sustainable urban development project in Copenhagen. Invoking Laurent Thévenot’s pragmatic sociology of ‘regimes of engagement’, we sketch a culturally sensitive approach to urban ecological activism, highlighting the critical moral capacities involved in building new forms of ‘commonality...

  18. Legume Logic & Green Manuring

    OpenAIRE

    Basavanagowda Nagabhushana, Nandeesh

    2014-01-01

    Brown plant hopper showed me the way into organic farming. In 2001, I started my practice with logic of legumes just to cut down the 45 percent expenses of my paddy on fertilizers, pesticides and herbicides. Later as I realized each and every plant carries it’s own nutrients, medicinal values and characters. Plants like millets, oil seeds, spices, di-cots, monocots and weeds all being used as a green manure. For all my agriculture problems and crop demands, I look for the answers only thro...

  19. Harmonic supergraphs. Green functions

    International Nuclear Information System (INIS)

    Galperin, A.; Ivanov, E.; Gievetsky, V.; Sokatchev, E.

    1985-01-01

    The quantization procedure in the harmonic superspace approach is worked out. Harmonic distributions are introduced and are used to construct the analytic superspace delta-functions and the Green functions for the hypermultiplet and the N=2 Yang-Mills superfields. The gauge fixing is described and the relevant Faddeev-Popov ghosts are defined. The corresponding BRST transformations are found. The harmonic superspace quantization of the N=2 gauge theory turns out to be rather simple and has many parallels with that for the standard (N=0) Yang-Mills theory. In particular, no ghosts-forghosts are needed

  20. Campaign Country Going Green?

    DEFF Research Database (Denmark)

    Poulsen, Bo

    2017-01-01

    justification. This paper finally discusses the reason for this greening of government initiated Danish energy saving campaigns, which is seen as an indirect result of the 1987 UN report, Our Common Future. The 1988 general election in Denmark led to the formation of a new center-right government coalition...... economics and not least a significant portion of patriotism. Environmental justification was almost entirely absent throughout the 1970s and 1980s. This changed only from 1989 onwards, as government initiatives to curb the ever rising consumption of energy commenced an extensive use of environmental...

  1. Green Consumption Behavior Antecedents

    DEFF Research Database (Denmark)

    Pagiaslis, Anastasios; Krystallis Krontalis, Athanasios

    2014-01-01

    The present study adds to the evolving literature on green consumer behavior by examining through statistically robust methods the effect and interrelationships of the key constructs of environmental concern, consumer environmental knowledge, beliefs about biofuels, and behavioral intention (i...... for the environment has a positive and direct impact on environmental knowledge, beliefs, and behavioral intention. Also, demographics determine levels of concern for the environment and environmental knowledge. All constructs associate positively with one another delineating that the interdependencies between them...... are important when accounting for environmental behavior. Future research should validate present results with the use of cross-cultural samples and investigate whether environmental concern increases due to social desirability response bias....

  2. Green Tourism Marketing Model1

    OpenAIRE

    Hasan, Ali

    2015-01-01

    Green Tourism Marketing Model research as efforts to develop environmentally friendly tourism destination, the synergy of government, business and community participation become the driving force of tourism product development with highly competitive. In the long term, this research aims to provide the marketing concept of green tourism as economic development efforts and strengthen the environment (eco-growth) through the development of green tourism marketing models. The ...

  3. The trashing of Big Green

    International Nuclear Information System (INIS)

    Felten, E.

    1990-01-01

    The Big Green initiative on California's ballot lost by a margin of 2-to-1. Green measures lost in five other states, shocking ecology-minded groups. According to the postmortem by environmentalists, Big Green was a victim of poor timing and big spending by the opposition. Now its supporters plan to break up the bill and try to pass some provisions in the Legislature

  4. The guide to greening cities

    National Research Council Canada - National Science Library

    Johnston, Sadhu Aufochs

    2013-01-01

    ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17 CHAPTER 3. Leading in the Community: Using City Assets, Policy, Partnerships, and Persuasion . . Case in Point: Returning to Green City Roots and Loving El...

  5. Toronto green roof construction standard

    International Nuclear Information System (INIS)

    Aster, D.

    2007-01-01

    Toronto City Council adopted a green roof strategy in February 2006. This paper reviewed the by-law governing the strategy as well as the work in progress to develop minimum standards for the design and construction of green roofs in Toronto. The strategy included a series of recommendations regarding the installation of green roofs on city buildings; a pilot grant program; using the development process to encourage green roofs; and, public education and promotion. It was noted that compared to Europe, the development of standards for green roofs in North America is in its early stages. As an emerging sustainable technology, there currently are no standards incorporated into Ontario's Building Code against which Toronto can measure the design and construction of green roofs. Therefore this paper included an analysis detailing how the recommended design requirements were able to support the City's green roof policy objectives and integrate the performance criteria for green roofs previously established and supported by Toronto City Council. The key policy objectives of the City's green roof strategy were to reduce the urban heat island effect; to address stormwater management implications in terms of quality and quantity; to improve the energy budgets of individual buildings; and, to improve air quality

  6. Product management in green markets

    Directory of Open Access Journals (Sweden)

    Čajka Zoran

    2005-01-01

    Full Text Available The present paper deals with the concept of green product management. To create a significantly greener economy, there will need to be a range of new and greener products and technologies. Today we are faced with a growth in more innovative "clean technology" solutions. Successful development of new green products requires high levels of communication and integration, good information, early consideration of green issues, support from top management, and benchmarking. The set of controllable tactical marketing tools (product, price, place and promotion that the company blends to produce the response it wants in the target green market, is the matter of the primary importance to the management.

  7. Ecological Transition and Green Investment

    International Nuclear Information System (INIS)

    Bureau, Dominique

    2016-01-01

    By introducing an exhaustible resource into an AK growth model, we can identify the determinants of the scale of green investment needs and its impact on capital accumulation dynamics. The role of green capital in the transition to a low-carbon economy depends, in particular, on the relative magnitudes of the elasticities of demand for polluting goods and of the substitutability between green capital and natural resources. The impact on the optimal savings rate also depends on the productivity of green capital and on the ability to adapt existing capital

  8. The Geography of Green Power

    Energy Technology Data Exchange (ETDEWEB)

    OShaughnessy, Eric J [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Heeter, Jenny S [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Volpi, Christina M [National Renewable Energy Laboratory (NREL), Golden, CO (United States)

    2017-10-25

    Green power refers to the voluntary purchase of renewable electricity by retail electricity customers. Green power is unlike compliance-based renewable energy procurement imposed by law or regulation. In 2016, over six million customers procured about 95 million megawatt-hours (MWh) of green power in the United States, which represents about 28% of all U.S. renewable energy sales, excluding large hydropower. In this fact sheet, we use available data to illustrate the geography of green power demand (in terms of number of customers) and supply (in terms of MWh of generation) by state.

  9. Can Growth Be Green?

    Science.gov (United States)

    Gough, Ian

    2015-01-01

    This short article, based on a presentation at the London School of Economics, criticizes the common opinion that "green growth" offers a relatively painless - some even say pain-free - transition path for capitalist economies. After a brief summary of the daunting arithmetic entailed in combining fast decarbonization with continuing growth, the article advances 3 propositions. First, market-based carbon mitigation programs, such as carbon trading, cannot be sufficient and must be coupled with other policy pillars that foster transformative investment and widespread regulation. Second, a political economy of climate policy needs to draw on the lessons of comparative social policy research, which emphasizes the role of international pressures, interests, institutions, and ideas. Taking these into account gives a more realistic perspective on climate policy making in today's neoliberal world. Third, more radical policies on both consumption and production are called for, to ensure that carbon mitigation is not pursued at the expense of equity and social welfare. These include policies to restrain high-carbon luxury consumption and a transition toward shorter paid working time. The conclusion is that a realistic program of green growth will be immensely difficult and entail radical political change. © SAGE Publications 2015.

  10. Greening of orthopedic surgery.

    Science.gov (United States)

    Lee, Rushyuan J; Mears, Simon C

    2012-06-01

    Every year, 4 billion pounds of waste are produced by health care facilities, and the amount continues to increase annually. In response, a movement toward greening health care has been building, with a particular focus on the operating room. Between 20% and 70% of health care waste originates from a hospital's operating room, and up to 90% of operating room waste is improperly sorted and sent for costly and unneeded hazardous waste processing. Recent successful changes include segregation of hospital waste, substitution of the ubiquitous polypropylene plastic wrap used for the sterilization and handling of surgical equipment with metal cases, and the reintroduction of reusable surgical gowns. Orthopedic-related changes include the successful reprocessing and reuse of external fixators, shavers, blades, burs, and tourniquets. These changes have been shown to be environmentally and economically beneficial. Early review indicates that these changes are feasible, but a need exists for further evaluation of the effect on the operating room and flow of the surgical procedure and of the risks to the surgeons and operating room staff. Other key considerations are the effects of reprocessed and reused equipment on patient care and outcome and the role of surgeons in helping patients make informed decisions regarding surgical care. The goals of this study were to summarize the amount and types of waste produced in hospitals and operating rooms, highlight the methods of disposal used, review disposal methods that have been developed to reduce waste and improve recycling, and explore future developments in greening health care. Copyright 2012, SLACK Incorporated.

  11. Green power certification: environmental and consumer protection benefits of the Green-e programme

    Energy Technology Data Exchange (ETDEWEB)

    Wingate, M.; Hamrin, J. [Center for Resource Solutions (United States); Rabago, K. [Rocky Mountain Inst. (United States); Wiser, R. [Lawrence Berkeley National Lab. (United States)

    2000-06-01

    This article gives details of the Green-e environmental certification programme which certifies electricity generated from renewable energy sources in the US. This first non-profit certification programme originally was set up for California, and has now spread to other regions. The objectives of the Green-e programme, the need for the electricity product to meet minimum criteria to qualify, marketer requirements, verification of product claims, administration of the programme, and the second year programme results are discussed. The way in which the Green-e programme fits in with other programmes such as those set up by the state and federal customer protection agencies to help consumers select environmentally superior power is described.

  12. Green Consumerism : an Eco-Friendly Behaviour Form Through The Green Product Consumption and Green Marketing

    Directory of Open Access Journals (Sweden)

    Wiwik Handayani

    2017-09-01

    Full Text Available This research is referred to analyse the influence of consumer attitude of green product towards purchase intention. The consumer attitude of green product is a psychological tendencies that is expressed by evaluating a certain entity with some advantage or disadvantage considerations. The problem of this research is the low of cunsumer awareness to consume green product, because the lack to comprehend the importance of green product usage for health and eco-friendly. The purpose of this research is to test the influence of consumer attitude of green products towards purchase intention. Hypothesis testing using Partial Least Square (PLS. The result of analysis show that there is influence among consumer attitude of green product towards consumer purchase intention significantly.

  13. The Green Experiment: Cities, Green Stormwater Infrastructure, and Sustainability

    OpenAIRE

    Christopher M. Chini; James F. Canning; Kelsey L. Schreiber; Joshua M. Peschel; Ashlynn S. Stillwell

    2017-01-01

    Green infrastructure is a unique combination of economic, social, and environmental goals and benefits that requires an adaptable framework for planning, implementing, and evaluating. In this study, we propose an experimental framework for policy, implementation, and subsequent evaluation of green stormwater infrastructure within the context of sociotechnical systems and urban experimentation. Sociotechnical systems describe the interaction of complex systems with quantitative and qualitative...

  14. Technological Advances in Huanglongbing (HLB or Citrus Greening Disease Management

    Directory of Open Access Journals (Sweden)

    Krishna Prasad Paudyal

    2015-12-01

    Full Text Available Huanglongbing (HLB, previously citrus greening disease, is the most destructive of citrus species causing major threat to the world citrus industry. The disease was reported from China in 1919 and now known to occur in more than 40 different countries of Asia, Africa, South and North America. Three species of gram negative bacterium namely Candidatus Liberibacter asiaticus, Candidatus Liberibacter africanus and Candidatus Liberibacter americanus are the casual organisms of HLB, respectively prevailing in the continent of Asia, Africa and South America. It is one of the most extensively researched subjects in citriculture world. HLB was detected in 2004 and 2005, respectively in San Paulo of Brazil and Florida of USA: the two leading citrus production hub of the world causing huge economic loss within 5 years of first detection. Since then research on HLB detection and management was further accelerated in American continents. This paper presents the scientific advancement made on detection, spread, economic losses caused by HLB in different parts of the world and controlling management strategies. Remarkable achievements have been made on HLB detection techniques including iodine test, qPCR and more recently in spectroscopy. While efforts are being made to develop resistance varieties using conventional and biotechnological tools management strategy which includes reduction of inoculums source, vector control and replant with disease-free planting materials still remains major option for HLB control. Citrus intercropping with guava have shown promising results for vector reduction.

  15. In situ evaluation of cadmium biomarkers in green algae

    Energy Technology Data Exchange (ETDEWEB)

    Simon, Dana F.; Davis, Thomas A. [Department of Chemistry, University of Montreal, P.O. Box 6128, Succursale Centre-ville, Montreal, Quebec H3C 3J7 (Canada); Tercier-Waeber, Mary-Lou [Analytical and Biophysical Environmental Chemistry, University of Geneva, Sciences II, 30 Quai Ernest-Ansermet, 1211 Geneva 4 (Switzerland); England, Roxane [Department of Chemistry, University of Montreal, P.O. Box 6128, Succursale Centre-ville, Montreal, Quebec H3C 3J7 (Canada); Wilkinson, Kevin J., E-mail: kj.wilkinson@umontreal.ca [Department of Chemistry, University of Montreal, P.O. Box 6128, Succursale Centre-ville, Montreal, Quebec H3C 3J7 (Canada)

    2011-10-15

    In situ measurements provide data that are the highly representative of the natural environment. In this paper, laboratory-determined biomarkers of Cd stress that were previously identified for the green alga Chlamydomonas reinhardtii, were tested in two French rivers: a contaminated site on the Riou Mort River and an 'uncontaminated' reference site on the Lot River. Transcript abundance levels were determined by real time qPCR for biomarkers thought to be Cd sensitive. Transcript levels were significantly higher (>5 fold) for organisms exposed to the contaminated site as compared to those exposed at the uncontaminated site. Biomarker mRNA levels were best correlated to free Cd (Cd{sup 2+}) rather than intracellular Cd, suggesting that they may be useful indicators of in situ stress. The paper shows that biomarker expression levels increased with time, were sensitive to metal levels and metal speciation and were higher in the 'contaminated' as opposed to the 'reference' site. - Highlights: > Biomarkers of Cd stress were tested in a contaminated and a reference site. > The organism was viable under exposure conditions and metal accumulation occurred. > Biomarker levels were correlated to Cd{sup 2+} and were higher in the contaminated site. - Algal transcription levels of several biomarkers were studied in two natural waters in situ.

  16. 77 FR 2296 - Office of Federal High-Performance Green Buildings; the Green Building Advisory Committee...

    Science.gov (United States)

    2012-01-17

    ... Federal High-Performance Green Buildings; the Green Building Advisory Committee; Notification of Upcoming... teleconference meetings of the Green Building Advisory Committee (the Committee). The teleconference meetings are... Federal High Performance Green Buildings, Office of Governmentwide Policy, General Services Administration...

  17. Use of next generation sequencing data to develop a qPCR method for specific detection of EU-unauthorized genetically modified Bacillus subtilis overproducing riboflavin.

    Science.gov (United States)

    Barbau-Piednoir, Elodie; De Keersmaecker, Sigrid C J; Delvoye, Maud; Gau, Céline; Philipp, Patrick; Roosens, Nancy H

    2015-11-11

    Recently, the presence of an unauthorized genetically modified (GM) Bacillus subtilis bacterium overproducing vitamin B2 in a feed additive was notified by the Rapid Alert System for Food and Feed (RASFF). This has demonstrated that a contamination by a GM micro-organism (GMM) may occur in feed additives and has confronted for the first time,the enforcement laboratories with this type of RASFF. As no sequence information of this GMM nor any specific detection or identification method was available, Next GenerationSequencing (NGS) was used to generate sequence information. However, NGS data analysis often requires appropriate tools, involving bioinformatics expertise which is not alwayspresent in the average enforcement laboratory. This hampers the use of this technology to rapidly obtain critical sequence information in order to be able to develop a specific qPCRdetection method. Data generated by NGS were exploited using a simple BLAST approach. A TaqMan® qPCR method was developed and tested on isolated bacterial strains and on the feed additive directly. In this study, a very simple strategy based on the common BLAST tools that can be used by any enforcement lab without profound bioinformatics expertise, was successfully used toanalyse the B. subtilis data generated by NGS. The results were used to design and assess a new TaqMan® qPCR method, specifically detecting this GM vitamin B2 overproducing bacterium. The method complies with EU critical performance parameters for specificity, sensitivity, PCR efficiency and repeatability. The VitB2-UGM method also could detect the B. subtilis strain in genomic DNA extracted from the feed additive, without prior culturing step. The proposed method, provides a crucial tool for specifically and rapidly identifying this unauthorized GM bacterium in food and feed additives by enforcement laboratories. Moreover, this work can be seen as a case study to substantiate how the use of NGS data can offer an added value to easily

  18. Characterization of relative abundance of lactic acid bacteria species in French organic sourdough by cultural, qPCR and MiSeq high-throughput sequencing methods.

    Science.gov (United States)

    Michel, Elisa; Monfort, Clarisse; Deffrasnes, Marion; Guezenec, Stéphane; Lhomme, Emilie; Barret, Matthieu; Sicard, Delphine; Dousset, Xavier; Onno, Bernard

    2016-12-19

    In order to contribute to the description of sourdough LAB composition, MiSeq sequencing and qPCR methods were performed in association with cultural methods. A panel of 16 French organic bakers and farmer-bakers were selected for this work. The lactic acid bacteria (LAB) diversity of their organic sourdoughs was investigated quantitatively and qualitatively combining (i) Lactobacillus sanfranciscensis-specific qPCR, (ii) global sequencing with MiSeq Illumina technology and (iii) molecular isolates identification. In addition, LAB and yeast enumeration, pH, Total Titratable Acidity, organic acids and bread specific volume were analyzed. Microbial and physico-chemical data were statistically treated by Principal Component Analysis (PCA) and Hierarchical Ascendant Classification (HAC). Total yeast counts were 6 log 10 to 7.6 log 10 CFU/g while LAB counts varied from 7.2 log 10 to 9.6 log 10 CFU/g. Values obtained by L. sanfranciscensis-specific qPCR were estimated between 7.2 and 10.3 log 10 CFU/g, except for one sample at 4.4 log 10 CFU/g. HAC and PCA clustered the sixteen sourdoughs into three classes described by their variables but without links to bakers' practices. L. sanfranciscensis was the dominant species in 13 of the 16 sourdoughs analyzed by Next Generation Sequencing (NGS), by the culture dependent method this species was dominant only in only 10 samples. Based on isolates identification, LAB diversity was higher for 7 sourdoughs with the recovery of L. curvatus, L. brevis, L. heilongjiangensis, L. xiangfangensis, L. koreensis, L. pontis, Weissella sp. and Pediococcus pentosaceus, as the most representative species. L. koreensis, L. heilongjiangensis and L. xiangfangensis were identified in traditional Asian food and here for the first time as dominant in organic sourdough. This study highlighted that L. sanfranciscensis was not the major species in 6/16 sourdough samples and that a relatively high LAB diversity can be observed in French organic

  19. The "Green" Root Beer Laboratory

    Science.gov (United States)

    Clary, Renee; Wandersee, James

    2010-01-01

    No, your students will not be drinking green root beer for St. Patrick's Day--this "green" root beer laboratory promotes environmental awareness in the science classroom, and provides a venue for some very sound science content! While many science classrooms incorporate root beer-brewing activities, the root beer lab presented in this article has…

  20. Green grabbing debate and Madagascar

    DEFF Research Database (Denmark)

    Casse, Thorkil; Razafy, Fara Lala; Wurtzebach, Zachary

    2017-01-01

    and capitalise natural assets. First, to provide some context on the green grabbing debate, we discuss the trade-offs between conservation and development objectives. In addition, we refer briefly to the broader land grabbing debate of which green grabbing is a sub-component. Second, we question the theoretical...

  1. Green investment: Trends and determinants

    International Nuclear Information System (INIS)

    Eyraud, Luc; Clements, Benedict; Wane, Abdoul

    2013-01-01

    This paper fills a gap in the macroeconomic literature on renewable sources of energy. It offers a definition of green investment and analyzes the trends and determinants of this investment over the last decade for 35 advanced and emerging countries. We use a new multi-country historical dataset and find that green investment has become a key driver of the energy sector and that its rapid growth is now mostly driven by China. Our econometric results suggest that green investment is boosted by economic growth, a sound financial system conducive to low interest rates, and high fuel prices. We also find that some policy interventions, such as the introduction of carbon pricing schemes or “feed-in-tariffs,” which require use of “green” energy, have a positive and significant impact on green investment. Other interventions, such as biofuel support, do not appear to be associated with higher green investment. - Highlights: • We offer a definition of green investment and review its trend since 2000. • We analyze its determinants from both theoretical and empirical perspectives. • Green investment is boosted by economic growth, interest rates, and fuel prices. • Feed-in-tariffs and carbon pricing schemes impact positively green investment

  2. Summary of Green Building Programs

    Energy Technology Data Exchange (ETDEWEB)

    2002-08-01

    In early 2002, the National Association of Home Builders completed a census of residential green building programs across the United States to assess differences and similarities among programs. This report catalogs different ways that builders participate in residential green building programs.

  3. Green accounts & day high schools

    DEFF Research Database (Denmark)

    Jørgensen, Michael Søgaard

    1997-01-01

    The arcticle presents the concept of green accounts and describes how it can be used in the daily work and the teaching at day high schools.......The arcticle presents the concept of green accounts and describes how it can be used in the daily work and the teaching at day high schools....

  4. Being Green: Managing Chemical Use

    DEFF Research Database (Denmark)

    Goodsite, Michael Evan; Warncke, Esbern; Moseholm, Lars

    2010-01-01

    The definitive, practical, go-to resource guide on helping all charities become more "green" Nonprofit Guide to Going Green is your comprehensive learning tool to guide nonprofits and NGOs towards becoming greener. A desktop reference for any charitable organization to become greener, this essent...

  5. Virtual Rewards for Driving Green

    Science.gov (United States)

    Pritchard, Josh

    2010-01-01

    Carbon dioxide from automobiles is a major contributor to global climate change. In "Virtual Rewards for Driving Green," Josh Pritchard proposes a computer application that will enable fuel-efficient drivers to earn "green" dollars with which to buy digital merchandise on the Web. Can getting items that exist only in cyberspace actually change a…

  6. Water for greening the economy

    CSIR Research Space (South Africa)

    De Lange, Willem J

    2015-03-01

    Full Text Available ] for such an overview). The focus here falls on a discussion aimed to improve alignment between water resource management and the principles of a green economy. Previous chapters have made it clear that a green economy requires a holistic approach towards policy...

  7. Sound absorption with green materials

    Science.gov (United States)

    Trematerra, Amelia; Lombardi, Ilaria

    2017-07-01

    Green materials are a valid alternative to traditional materials that are by-products of processing oil. At the end of their useful life, green materials can be disposed of without polluting the environment. They are now being used in the construction and automotive industries. While, studies are currently being carried out in the aviation sector on the use of green materials for non-structural components of airplanes. Green materials can be used to improve the acoustic comfort inside buildings as well as mitigate reverberation, echoes effects and reduce the transmission of noise between rooms. In this paper, the acoustic measurements of the properties of green materials are reported. The absorption coefficient of samples of the materials were measured in the frequency range from 200 Hz to 2,000 Hz with an impedance tube, with the flow resistance being measured.

  8. World green electricity, sustaining investments

    International Nuclear Information System (INIS)

    Le Jannic, N.

    2013-01-01

    The contribution of the green production to the world production of electricity reached 20.2% in 2011, it means a slight increase in respect to the figure of 2010: 19.8%. Green electricity is the second source of electricity behind fossil energy (67.9%) but before nuclear power (11.7%). The decrease in nuclear power due to the Fukushima accident has automatically benefited green electricity. The figures show the importance of China, China is now the first electricity producer in the world before US and also passed US for the production of green electricity. At the world scale the production of green electricity can break down into: hydro energy (80.5%), wind energy (10.3%), biomass (6.2%), geothermal energy (1.6%) and solar energy (1.4%). The crisis has slowed down the investment in renewable energies in Europe. (A.C.)

  9. Future Green Buildings

    DEFF Research Database (Denmark)

    Mathiesen, Brian Vad; Drysdale, David; Lund, Henrik

    an energy system integration perspective, heat savings, electricity savings, and user behavioural aspects as well as energy storage and household level flexibility. Many reports on green or sustainable buildings focus only on savings levels and disregard the cost of renewable energy production. Some reports......Efficient buildings are essential for an affordable Danish energy supply in 2050. The purpose of this report is to describe the contribution and role of the building sector in a 100% renewable energy future, as well as the transitions that are necessary in the building sector to support this change....... The report builds on a literature review encompassing more than 50 reports and research papers over the last 10 years and more than a two decades knowledge about the interactions between different components of the energy sector. The review has been focused on aspects such as cost-effective solutions from...

  10. Green Maritime Logistics

    DEFF Research Database (Denmark)

    Psaraftis, Harilaos N.

    2014-01-01

    Typical problems in maritime logistics include, among others, optimal ship speed, ship routing and scheduling, fleet deployment, fleet size and mix, weather routing, intermodal network design, modal split, transshipment, queuing at ports, terminal management, berth allocation, and total supply...... chain management. The traditional analysis of these problems has been in terms of cost- benefit and other optimization criteria from the point of view of the logistics provider, carrier, shipper, or other end-user. Such traditional analysis by and large either ignores environmental issues, or considers...... them of secondary importance. Green maritime logistics tries to bring the environmental dimension into the problem, and specifically the dimension of emissions reduction, by analyzing various trade-offs and exploring ‘win-win’ solutions. This talk takes a look at the trade-offs that are at stake...

  11. Green symbiotic cloud communications

    CERN Document Server

    Mustafa, H D; Desai, Uday B; Baveja, Brij Mohan

    2017-01-01

    This book intends to change the perception of modern day telecommunications. Communication systems, usually perceived as “dumb pipes”, carrying information / data from one point to another, are evolved into intelligently communicating smart systems. The book introduces a new field of cloud communications. The concept, theory, and architecture of this new field of cloud communications are discussed. The book lays down nine design postulates that form the basis of the development of a first of its kind cloud communication paradigm entitled Green Symbiotic Cloud Communications or GSCC. The proposed design postulates are formulated in a generic way to form the backbone for development of systems and technologies of the future. The book can be used to develop courses that serve as an essential part of graduate curriculum in computer science and electrical engineering. Such courses can be independent or part of high-level research courses. The book will also be of interest to a wide range of readers including b...

  12. Markets: green utilities

    International Nuclear Information System (INIS)

    Wood, Elisa

    2006-01-01

    Publicly owned utilities have consistently led the United States in the rate of customer participation in green power programmes. The US has about 2000 community and state-owned utilities, which serve 43 million customers and account for about 16.6% of kilowatt-hour sales to consumers. In all, public power is responsible for about 10% of the nation's installed electric capacity. Investor owned utilities account for 39%, with the remainder of the nation's power mostly from independent power generators. Although IOUs have almost four times as much electric capacity as public power, they edge out public power by only a small margin when it comes to renewable capacity. IOUs are responsible for 24,577.5 MW of renewable capacity, compared to the 21,338 MW installed by public power. The reasons discussed by the author range from small town advantage to clean and cheap power. (Author)

  13. Comparison of bacterial culture and qPCR testing of rectal and pen floor samples as diagnostic approaches to detect enterotoxic Escherichia coli in nursery pigs

    DEFF Research Database (Denmark)

    Weber, N. R.; Nielsen, J. P.; Hjulsager, Charlotte Kristiane

    2017-01-01

    Enterotoxigenic E. coli (ETEC) are a major cause of diarrhoea in weaned pigs. The objective of this study was to evaluate the agreement at pen level among three different diagnostic approaches for the detection of ETEC in groups of nursery pigs with diarrhoea. The diagnostic approaches used were......: bacterial culturing of faecal samples from three pigs (per pen) with clinical diarrhoea and subsequent testing for virulence genes in E. coli isolates; bacterial culturing of pen floor samples and subsequent testing for virulence genes in E. coli isolates; qPCR testing of pen floor samples in order...... to determine the quantity of F18 and F4 genes. The study was carried out in three Danish pig herds and included 31 pens with a pen-level diarrhoea prevalence of > 25%, as well as samples from 93 diarrhoeic nursery pigs from these pens. All E. coli isolates were analysed by PCR and classified as ETEC when genes...

  14. Comparison of individual and pooled samples for quantification of antimicrobial resistance genes in swine feces by high-throughput qPCR

    DEFF Research Database (Denmark)

    Clasen, Julie; Mellerup, Anders; Olsen, J. E.

    2015-01-01

    There is a considerable societal interest in the careful monitoring of antimicrobial resistance (AMR) levels in human and animal populations. Sampling and data analysis can be both costly and time consuming. Optimization of sample pooling procedures is therefore important to reduce costs...... and analysis times. The objective of this study was to estimate how many individual fecal samples are needed to pool to get a representative sample for quantification of AMR-genes in a Danish pig herd. 20 individual fecal samples were collected from one section in a Danish pig herd. One to five rectal fecal...... samples were taken from each pen with respect to the number of pigs in the pen. A total of 48 pools were made of increasing number of individual samples. The levels of 9 different AMR-genes were quantified using dynamic qPCR arrays on the BioMark HD system(Fluidigm®).DNA was extracted using the Maxwell...

  15. Green Roofs for Stormwater Runoff Control - Abstract

    Science.gov (United States)

    This project evaluated green roofs as a stormwater management tool. Specifically, runoff quantity and quality from green and flat asphalt roofs were compared. Evapotranspiration from planted green roofs and evaporation from unplanted media roofs were also compared. The influence...

  16. Green chemistry at work

    Energy Technology Data Exchange (ETDEWEB)

    Frost, J. [Michigan State Univ., East Lansing, MI (United States)

    1994-12-31

    The 1.7 billion pounds of benzene produced each year in the US provide one measure of its utility. At the same time, there are a number of environmental reasons for avoiding the use of benzene in chemical manufacture. Perhaps most compelling: benzene is a potent carcinogen. Scrutiny of many of the chemicals derived from benzene reveals that each molecule contains at least one oxygen atom while benzene completely lacks oxygen atoms. Introduction of oxygen to make up for this lack can require processes that are environmentally problematic. One of the steps used to introduce oxygen atoms during manufacture of adipic acid, a component of Nylon 66, is responsible for 10% of the annual global increase in atmospheric nitrous oxide. This by-product is a causative agent of atmospheric ozone depletion and has been implicated in global warming. With support from EPA and the National Science Foundation, alternative manufacturing processes are being explored. By these new methods, chemicals usually created from benzene are made instead from nontoxic glucose, a component of table sugar. Unlike benzene, glucose is obtained from such renewable resources as plant starch and cellulose. ``Green`` manufacturing routes ideally should lead to chemicals that are economically competitive with chemicals produced by traditional methods. For two chemicals of roughly comparable cost, the consumer or producer can then be realistically expected to choose in favor of the chemical produced by a ``green`` process. Projections indicate that catechol and hydroquinone can be biocatalytically produced from glucose at a cost competitive with current market prices. Synthesis of chemicals from glucose using biocatalysis offers the premise of achieving fundamental environmental improvement while increasing the demand for agricultural products. In the final analysis, what is good for the environment can also be good for American agriculture.

  17. Urban Green Infrastructure: German Experience

    Directory of Open Access Journals (Sweden)

    Diana Olegovna Dushkova

    2016-06-01

    Full Text Available The paper presents a concept of urban green infrastructure and analyzes the features of its implementation in the urban development programmes of German cities. We analyzed the most shared articles devoted to the urban green infrastructure to see different approaches to definition of this term. It is based on materials of field research in the cities of Berlin and Leipzig in 2014-2015, international and national scientific publications. During the process of preparing the paper, consultations have been held with experts from scientific institutions and Administrations of Berlin and Leipzig as well as local experts from environmental organizations of both cities. Using the German cities of Berlin and Leipzig as examples, this paper identifies how the concept can be implemented in the program of urban development. It presents the main elements of green city model, which include mitigation of negative anthropogenic impact on the environment under the framework of urban sustainable development. Essential part of it is a complex ecological policy as a major necessary tool for the implementation of the green urban infrastructure concept. This ecological policy should embody not only some ecological measurements, but also a greening of all urban infrastructure elements as well as implementation of sustainable living with a greater awareness of the resources, which are used in everyday life, and development of environmental thinking among urban citizens. Urban green infrastructure is a unity of four main components: green building, green transportation, eco-friendly waste management, green transport routes and ecological corridors. Experience in the development of urban green infrastructure in Germany can be useful to improve the environmental situation in Russian cities.

  18. A randomized and blinded comparison of qPCR and NGS-based detection of aneuploidy in a cell line mixture model of blastocyst biopsy mosaicism.

    Science.gov (United States)

    Goodrich, David; Tao, Xin; Bohrer, Chelsea; Lonczak, Agnieszka; Xing, Tongji; Zimmerman, Rebekah; Zhan, Yiping; Scott, Richard T; Treff, Nathan R

    2016-11-01

    A subset of preimplantation stage embryos may possess mosaicism of chromosomal constitution, representing a possible limitation to the clinical predictive value of comprehensive chromosome screening (CCS) from a single biopsy. However, contemporary methods of CCS may be capable of predicting mosaicism in the blastocyst by detecting intermediate levels of aneuploidy within a trophectoderm biopsy. This study evaluates the sensitivity and specificity of aneuploidy detection by two CCS platforms using a cell line mixture model of a mosaic trophectoderm biopsy. Four cell lines with known karyotypes were obtained and mixed together at specific ratios of six total cells (0:6, 1:5, 2:4, 3:3, 4:2, 5:1, and 6:0). A female euploid and a male trisomy 18 cell line were used for one set, and a male trisomy 13 and a male trisomy 15 cell line were used for another. Replicates of each mixture were prepared, randomized, and blinded for analysis by one of two CCS platforms (quantitative polymerase chain reaction (qPCR) or VeriSeq next-generation sequencing (NGS)). Sensitivity and specificity of aneuploidy detection at each level of mosaicism was determined and compared between platforms. With the default settings for each platform, the sensitivity of qPCR and NGS were not statistically different, and 100 % specificity was observed (no false positives) at all levels of mosaicism. However, the use of previously published custom criteria for NGS increased sensitivity but also significantly decreased specificity (33 % false-positive prediction of aneuploidy). By demonstrating increased false-positive diagnoses when reducing the stringency of predicting an abnormality, these data illustrate the importance of preclinical evaluation of new testing paradigms before clinical implementation.

  19. Validating Internal Control Genes for the Accurate Normalization of qPCR Expression Analysis of the Novel Model Plant Setaria viridis.

    Directory of Open Access Journals (Sweden)

    Julia Lambret-Frotté

    Full Text Available Employing reference genes to normalize the data generated with quantitative PCR (qPCR can increase the accuracy and reliability of this method. Previous results have shown that no single housekeeping gene can be universally applied to all experiments. Thus, the identification of a suitable reference gene represents a critical step of any qPCR analysis. Setaria viridis has recently been proposed as a model system for the study of Panicoid grasses, a crop family of major agronomic importance. Therefore, this paper aims to identify suitable S. viridis reference genes that can enhance the analysis of gene expression in this novel model plant. The first aim of this study was the identification of a suitable RNA extraction method that could retrieve a high quality and yield of RNA. After this, two distinct algorithms were used to assess the gene expression of fifteen different candidate genes in eighteen different samples, which were divided into two major datasets, the developmental and the leaf gradient. The best-ranked pair of reference genes from the developmental dataset included genes that encoded a phosphoglucomutase and a folylpolyglutamate synthase; genes that encoded a cullin and the same phosphoglucomutase as above were the most stable genes in the leaf gradient dataset. Additionally, the expression pattern of two target genes, a SvAP3/PI MADS-box transcription factor and the carbon-fixation enzyme PEPC, were assessed to illustrate the reliability of the chosen reference genes. This study has shown that novel reference genes may perform better than traditional housekeeping genes, a phenomenon which has been previously reported. These results illustrate the importance of carefully validating reference gene candidates for each experimental set before employing them as universal standards. Additionally, the robustness of the expression of the target genes may increase the utility of S. viridis as a model for Panicoid grasses.

  20. Whole Genome Sequencing and Multiplex qPCR Methods to Identify Campylobacter jejuni Encoding cst-II or cst-III Sialyltransferase

    Directory of Open Access Journals (Sweden)

    Jason M. Neal-McKinney

    2018-03-01

    Full Text Available Campylobacter jejuni causes more than 2 million cases of gastroenteritis annually in the United States, and is also linked to the autoimmune sequelae Guillan–Barre syndrome (GBS. GBS often results in flaccid paralysis, as the myelin sheaths of nerve cells are degraded by the adaptive immune response. Certain strains of C. jejuni modify their lipooligosaccharide (LOS with the addition of neuraminic acid, resulting in LOS moieties that are structurally similar to gangliosides present on nerve cells. This can trigger GBS in a susceptible host, as antibodies generated against C. jejuni can cross-react with gangliosides, leading to demyelination of nerves and a loss of signal transduction. The goal of this study was to develop a quantitative PCR (qPCR method and use whole genome sequencing data to detect the Campylobactersialyltransferase (cst genes responsible for the addition of neuraminic acid to LOS. The qPCR method was used to screen a library of 89 C. jejuni field samples collected by the Food and Drug Administration Pacific Northwest Lab (PNL as well as clinical isolates transferred to PNL. In silico analysis was used to screen 827 C. jejuni genomes in the FDA GenomeTrakr SRA database. The results indicate that a majority of C. jejuni strains could produce LOS with ganglioside mimicry, as 43.8% of PNL isolates and 46.9% of the GenomeTrakr isolates lacked the cst genes. The methods described in this study can be used by public health laboratories to rapidly determine whether a C. jejuni isolate has the potential to induce GBS. Based on these results, a majority of C. jejuni in the PNL collection and submitted to GenomeTrakr have the potential to produce LOS that mimics human gangliosides.

  1. Use of high throughput qPCR screening to rapidly clone low frequency tumour specific T-cells from peripheral blood for adoptive immunotherapy

    Directory of Open Access Journals (Sweden)

    Serrano Oscar K

    2008-10-01

    Full Text Available Abstract Background The adoptive transfer of autologous tumor reactive lymphocytes can mediate significant tumor regression in some patients with refractory metastatic cancer. However, a significant obstacle for this promising therapy has been the availability of highly efficient methods to rapidly isolate and expand a variety of potentially rare tumor reactive lymphocytes from the natural repertoire of cancer patients. Methods We developed a novel in vitro T cell cloning methodology using high throughput quantitative RT-PCR (qPCR assay as a rapid functional screen to detect and facilitate the limiting dilution cloning of a variety of low frequency T cells from bulk PBMC. In preclinical studies, this strategy was applied to the isolation and expansion of gp100 specific CD8+ T cell clones from the peripheral blood of melanoma patients. Results In optimization studies, the qPCR assay could detect the reactivity of 1 antigen specific T cell in 100,000 background cells. When applied to short term sensitized PBMC microcultures, this assay could detect T cell reactivity against a variety of known melanoma tumor epitopes. This screening was combined with early limiting dilution cloning to rapidly isolate gp100154–162 reactive CD8+ T cell clones. These clones were highly avid against peptide pulsed targets and melanoma tumor lines. They had an effector memory phenotype and showed significant proliferative capacity to reach cell numbers appropriate for adoptive transfer trials (~1010 cells. Conclusion This report describes a novel high efficiency strategy to clone tumor reactive T cells from peripheral blood for use in adoptive immunotherapy.

  2. Microarray detection and qPCR screening of potential biomarkers of Folsomia candida (Collembola: Isotomidae) exposed to Bt proteins (Cry1Ab and Cry1Ac)

    International Nuclear Information System (INIS)

    Yuan, Yiyang; Krogh, Paul Henning; Bai, Xue; Roelofs, Dick; Chen, Fajun; Zhu-Salzman, Keyan; Liang, Yuyong; Sun, Yucheng; Ge, Feng

    2014-01-01

    The impact of Bt proteins on non-target arthropods is less understood than their effects on target organisms where the mechanism of toxic action is known. Here, we report the effects of two Bt proteins, Cry1Ab and Cry1Ac, on gene expression in the non-target collembolan, Folsomia candida. A customized microarray was used to study gene expression in F. candida specimens that were exposed to Cry1Ab and Cry1Ac. All selected transcripts were subsequently confirmed by qPCR. Eleven transcripts were finally verified, and three of them were annotated. The responses of all eleven transcripts were tested in specimens for both Cry1Ab and Cry1Ac at a series of concentrations. These transcripts were separated into two and three groups for Cry1Ab and Cry1Ac, respectively, depend on their expression levels. However, those eleven transcripts did not respond to the Bt proteins in Bt-rice residues. -- Highlights: • We examined the effects of Bt proteins on gene expression of Folsomia candida. • Eleven transcripts were up-regulated by Bt proteins (Cry1Ab and Cry1Ac). • Only three of the eleven transcripts were annotated. • The responses of 11 transcripts were tested on both Cry1Ab and Cry1Ac. • These transcripts did not respond to the Bt proteins in Bt-rice residues. -- Eleven potential molecular biomarkers of Folsomia candida to Cry1Ab and Cry1Ac were screened by microarray and qPCR analysis

  3. Going Green: The Business Case for Greening your Energy Company

    Energy Technology Data Exchange (ETDEWEB)

    Lavery, Greg

    2007-07-01

    We are all familiar with the challenges facing the energy industry: supply security, climate change, emerging cleaner technologies, retail competition, staffing, and the quest for growth. This paper demonstrates how a proactive environmentally considered ('green') corporate approach addresses these issues and unlocks four tangible areas of value addition for energy companies. Based on over a decade of experience by the author in this emerging field, this paper provides some golden rules for companies considering the green approach and showcases an Australian market leading energy company who is unlocking green value. (auth)

  4. Green Building Pro-Environment Behaviors: Are Green Users Also Green Buyers?

    Directory of Open Access Journals (Sweden)

    Xiaohuan Xie

    2017-09-01

    Full Text Available Pro-environment behaviors play a key role in advancing the development of green buildings. This study investigated the link between two green building pro-environment behaviors that require dissimilar resources: energy savings that do not require money in order to be more environmentally friendly and willingness to pay that involves economic resources including spending money in order to be more environmentally friendly. This study points out that the two pro-environment behaviors can be positively linked to each other. People who behave in an environmentally friendly manner at work would also be likely to pay an extra cost for a green building when buying a new home. The consistency of the two pro-environment behaviors can be explained by their common environmental beliefs: limits to growth and eco-crisis. The green building movement should prioritize pro-environmental behaviors and associated environmental beliefs to support green building policies, guidelines, and tools.

  5. Green materials for sustainable development

    Science.gov (United States)

    Purwasasmita, B. S.

    2017-03-01

    Sustainable development is an integrity of multidiscipline concept combining ecological, social and economic aspects to construct a liveable human living system. The sustainable development can be support through the development of green materials. Green materials offers a unique characteristic and properties including abundant in nature, less toxic, economically affordable and versatility in term of physical and chemical properties. Green materials can be applied for a numerous field in science and technology applications including for energy, building, construction and infrastructures, materials science and engineering applications and pollution management and technology. For instance, green materials can be developed as a source for energy production. Green materials including biomass-based source can be developed as a source for biodiesel and bioethanol production. Biomass-based materials also can be transformed into advanced functionalized materials for advanced bio-applications such as the transformation of chitin into chitosan which further used for biomedicine, biomaterials and tissue engineering applications. Recently, cellulose-based material and lignocellulose-based materials as a source for the developing functional materials attracted the potential prospect for biomaterials, reinforcing materials and nanotechnology. Furthermore, the development of pigment materials has gaining interest by using the green materials as a source due to their unique properties. Eventually, Indonesia as a large country with a large biodiversity can enhance the development of green material to strengthen our nation competitiveness and develop the materials technology for the future.

  6. Defining and certifying green power

    International Nuclear Information System (INIS)

    1998-02-01

    Studies have shown that as electric utilities restructure from monopolistic utilities to competitive open access retailers, there is an increasing demand by individual and institutional customers for green power. In the United States, 17 electricity suppliers have offered customers the opportunity to buy energy generated from renewable sources such as photovoltaic panels, wind turbines and biomass. Twenty other utilities are conducting market research in preparation for offering a similar program. It was suggested that in order to help the customers make their choice based on accurate information, generating facilities should be obligated to provide credible information about the environmental performance of electricity supply through standardized environmental profile labels. A list of agreed upon environmental indicators and performance levels must be established so that the 'environmental friendliness' of different generating facilities can be measured. One of the problems in tackling this issue is that there is disagreement about what constitutes green power. Opinions range from wind and solar generation being the only two forms of green power, to including even natural gas and nuclear energy (i.e. under the right conditions). The two programs that are used for the certification of green power in Canada and the United States are Canada's Environmental Choice Program and California's Green-e Renewable Electricity Branding Program. This report describes the two programs and summarizes the results of interviews conducted on the definition and certification of green power. 15 refs

  7. It's hard to be green: Reverse green value chain.

    Science.gov (United States)

    Couto, João; Tiago, Teresa; Gil, Artur; Tiago, Flávio; Faria, Sandra

    2016-08-01

    Firms have recently discovered that it is not enough to optimize internal processes and relationships with partners along the value chain to create a sustainable competitive market position. A clear customer orientation, which acknowledges that consumer buying behavior is complex and includes many elements implied in the value chain, is required. As companies offering green products are no exception to this rule, this study analyzes consumer behavior in Europe from a reserve green supply chain management perspective, using descriptive analyses and a structural equation model, with data collected by Flash Barometer comprising 26,573 responses from 28 European countries. The results suggest that European consumers are conscious of the green concept, but are not willing to buy or pay more for these products since the value is unclear. Companies offering green products must therefore rethink their strategies, especially in terms of value proposition, communication strategies, and eco-labeling. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. Quantification of massively parallel sequencing libraries - a comparative study of eight methods

    DEFF Research Database (Denmark)

    Hussing, Christian; Kampmann, Marie-Louise; Mogensen, Helle Smidt

    2018-01-01

    Quantification of massively parallel sequencing libraries is important for acquisition of monoclonal beads or clusters prior to clonal amplification and to avoid large variations in library coverage when multiple samples are included in one sequencing analysis. No gold standard for quantification...... estimates followed by Qubit and electrophoresis-based instruments (Bioanalyzer, TapeStation, GX Touch, and Fragment Analyzer), while SYBR Green and TaqMan based qPCR assays gave the lowest estimates. qPCR gave more accurate predictions of sequencing coverage than Qubit and TapeStation did. Costs, time......-consumption, workflow simplicity, and ability to quantify multiple samples are discussed. Technical specifications, advantages, and disadvantages of the various methods are pointed out....

  9. Green's functions in quantum physics

    CERN Document Server

    Economou, Eleftherios N

    2006-01-01

    The main part of this book is devoted to the simplest kind of Green's functions, namely the solutions of linear differential equations with a -function source. It is shown that these familiar Green's functions are a powerful tool for obtaining relatively simple and general solutions of basic problems such as scattering and bound-level information. The bound-level treatment gives a clear physical understanding of "difficult" questions such as superconductivity, the Kondo effect, and, to a lesser degree, disorder-induced localization. The more advanced subject of many-body Green's functions is presented in the last part of the book.

  10. Green Energy - Fiction and Reality

    Science.gov (United States)

    Cranberg, Lawrence

    2009-03-01

    The term ``Green Energy'' has been popularized to refer to energy sources that do not depend on fossil fuels. The oldest truly ``green'' energy source is wood fuel derived by photosynthesis for thermal comfort in the cold season. Thermal energy from the combustion of wood for personal heating in our 41 million fireplaces has greatly declined, due to an ``Anti-fireplace Hoax'' (1) that fireplaces are ``energy counterproductive.'' Physicists have a special obligation to address the problem that our major true source of ``Green Energy'' is widely misrepresented and neglected. 1. L. Cranberg, The Physics Teacher, Letter, January,l989

  11. Catalysis and sustainable (green) chemistry

    Energy Technology Data Exchange (ETDEWEB)

    Centi, Gabriele; Perathoner, Siglinda [Dipartimento di Chimica Industriale ed Ingegneria dei Materiali, University of Messina, Salita Sperone 31, 98166 Messina (Italy)

    2003-01-15

    Catalysis is a key technology to achieve the objectives of sustainable (green) chemistry. After introducing the concepts of sustainable (green) chemistry and a brief assessment of new sustainable chemical technologies, the relationship between catalysis and sustainable (green) chemistry is discussed and illustrated via an analysis of some selected and relevant examples. Emphasis is also given to the concept of catalytic technologies for scaling-down chemical processes, in order to develop sustainable production processes which reduce the impact on the environment to an acceptable level that allows self-depuration processes of the living environment.

  12. Green Jobs in Tennessee: Economic Impact of Green Investments

    OpenAIRE

    Murat Arik

    2011-01-01

    The term green jobs has been widely used to describe jobs in businesses that are particularly related to renewable energy, energy efficiency, or environmental sustainability. The Business and Economic Research Center has partnered with the Tennessee Department of Labor and Workforce Development to estimate the economic impact of six ground-breaking green investments in Tennessee: Hemlock Semiconductor, Wacker Chemie AG, Volkswagen, Nissan Leaf and Storage Battery Manufacturing, Tennessee Sola...

  13. Green roofs: potential at LANL

    Energy Technology Data Exchange (ETDEWEB)

    Pacheco, Elena M [Los Alamos National Laboratory

    2009-01-01

    Green roofs, roof systems that support vegetation, are rapidly becoming one of the most popular sustainable methods to combat urban environmental problems in North America. An extensive list of literature has been published in the past three decades recording the ecological benefits of green roofs; and now those benefits have been measured in enumerated data as a means to analyze the costs and returns of green roof technology. Most recently several studies have made substantial progress quantifying the monetary savings associated with storm water mitigation, the lessoning of the Urban Heat Island, and reduction of building cooling demands due to the implementation of green roof systems. Like any natural vegetation, a green roof is capable of absorbing the precipitation that falls on it. This capability has shown to significantly decrease the amount of storm water runoff produced by buildings as well as slow the rate at which runoff is dispensed. As a result of this reduction in volume and velocity, storm drains and sewage systems are relieved of any excess stress they might experience in a storm. For many municipalities and private building owners, any increase in storm water mitigation can result in major tax incentives and revenue that does not have to be spent on extra water treatments. Along with absorption of water, vegetation on green roofs is also capable of transpiration, the process by which moisture is evaporated into the air to cool ambient temperatures. This natural process aims to minimize the Urban Heat Island Effect, a phenomenon brought on by the dark and paved surfaces that increases air temperatures in urban cores. As the sun distributes solar radiation over a city's area, dark surfaces such as bitumen rooftops absorb solar rays and their heat. That heat is later released during the evening hours and the ambient temperatures do not cool as they normally would, creating an island of constant heat. Such excessively high temperatures induce heat

  14. Progress report on Green Deals 2013; Voortgangsrapportage Green Deals 2013

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2013-11-15

    In the Dutch governmental coalition agreement the Green Deal approach was announced in the autumn of 2010. The focus of the Green Deals is for people and companies to develop sustainable initiatives that contribute to economic growth. The Green Deal approach started with the theme energy, but has been extended with the themes biobased economy, climate, resources, buildings, food, mobility, water and biological diversity. This progress report provides an overview of the deals that this bottom-up approach has yielded. The report also provides information on the progress of the deals and the interim results of the approach and the individual deals. Also attention is paid to how the 146 Green Deals score on innovation and entrepreneurship [Dutch] In het regeerakkoord van het kabinet is in het najaar van 2010 de Green Deal-aanpak aangekondigd. Centraal in de aanpak staat dat mensen en bedrijven zoveel mogelijk ruimte krijgen voor eigen duurzame initiatieven die bijdragen aan economische groei. De aanpak is gestart vanuit het thema energie, maar beslaat inmiddels ook de thema's biobased economy, klimaat, grondstoffen, bouw, voedsel, mobiliteit, water en biodiversiteit. Deze voortgangsrapportage geeft een overzicht van de deals die deze bottom-up aanpak heeft opgeleverd. De rapportage informeert bovendien over de voortgang van de deals en over de tussentijdse resultaten van zowel de aanpak als de afzonderlijke deals. Ook wordt gekeken hoe de 146 Green Deals scoren op innovatief vermogen en ondernemerschap.

  15. Green Care Farms

    Directory of Open Access Journals (Sweden)

    Simone R. de Bruin PhD

    2015-10-01

    Full Text Available Objective: To explore the value of day services at green care farms (GCFs in terms of social participation for people with dementia. Method: Semi-structured interviews were conducted with people with dementia who attended day services at a GCF (GCF group, n = 21, were on a waiting list (WL for day services at a GCF (WL group, n = 12, or attended day services in a regular day care facility (RDCF group, n = 17 and with their family caregivers. Results: People with dementia in the GCF and WL group were primarily males, with an average age of 71 and 76 years, respectively, who almost all had a spousal caregiver. People with dementia in the RDCF group were mostly females with an average age of 85 years, most of whom had a non-spousal caregiver. For both the GCF and RDCF groups, it was indicated that day services made people with dementia feel part of society. The most important domains of social participation addressed by RDCFs were social interactions and recreational activities. GCFs additionally addressed the domains “paid employment” and “volunteer work.” Conclusion: GCFs are valuable in terms of social participation for a particular group of people with dementia. Matching characteristics of adult day services (ADS centers to the preferences and capacities of people with dementia is of importance. Diversity in ADS centers is therefore desirable.

  16. Greening the Judiciary

    Directory of Open Access Journals (Sweden)

    M Kidd

    2006-12-01

    Full Text Available Much of South Africa’s environmental law is relatively new. Most of South Africa’s judges received their formal legal educations before promulgation of the major part of our environmental law and almost certainly before environmental law was taught at universities. In recent years, there have been increasing instances of cases involving environmental matters coming to the courts. How are judges performing in these cases? It would appear that the judges’ performance is rather 'chequered' in environmental cases, which suggests that the judiciary needs to become more attuned to environmental law. I call this process, for purposes of this note, ‘greening the judiciary’. What I mean by this is not that judges must decide all environmental cases in a way that favours the environment, but that they must correctly consider, interpret and apply the relevant environmental law, and give environmental considerations appropriate deliberation. This note aims to identify, in admittedly somewhat general terms, the current state of environmental decision-making by judges and to suggest what needs to happen for such decisions to be improved.

  17. Renewing our green spaces

    CERN Multimedia

    CERN Bulletin

    2010-01-01

    CERN’s poplars were planted 50 years ago to soak up the surplus water in the ground beneath the Laboratory’s sites. Now that they have reached the end of their life-cycle, some of the poplars are in danger of falling or losing their dead branches. On Saturday, 17 April, as part of the campaign launched in February at Prévessin, work will start on replacing the poplars on central areas of the Meyrin site.   Replanting plan for the Cedars and kindergarten carparks. In July 2009, CERN was awarded an environmental label for its protection of rare flower species and the natural landscaping approach it has taken at its Meyrin site. The Laboratory has a very strict environmental policy: for years those in charge of CERN's green areas have given priority to natural management methods and have avoided the use of pesticides. In addition, patches of land are left unmowed in spring, allowing the local flora—especially the orchids—to grow naturally. Since l...

  18. Green channel cargo inspection system

    International Nuclear Information System (INIS)

    Shi Yuanping; Yu Jingsheng; Sun Hongqiang; Hao Pu; Cai Wenxia

    2011-01-01

    A radiation detection device was installed in the lanes of a highway toll station, radioactive rays which was collimated emitted through the measured, and arrived the detector. The average density of the fresh agricultural products belonged to Green channel and other prohibited items vary greatly, the absorption of radiation are different between the Green Channel Cargo and other substances. Prior to the experimental group, different standard samples which represent different models and goods were measured, the different standard samples were stored in a computer database. When the trucks get through the Green Channel, the detector will detect the radiation signal and bring to the computer, the computer will process the measured data, and make a conclusion whether the goods are Green Channel cargo. (authors)

  19. Green Building and School Construction.

    Science.gov (United States)

    DiNola, Ralph; Guerra, Jerry

    2002-01-01

    Discusses the benefits of green, or high-performance, buildings, such as health and comfort, cost effectiveness, and sustainability. Explores the barriers to their use by schools--most notably cost. Offers suggestions on overcoming these barriers. (EV)

  20. Green Power Partnership Eligible Organizations

    Science.gov (United States)

    The U.S. EPA's Green Power Partnership is a voluntary partnership program designed to reduce the environmental impact of electricity generation by promoting renewable energy. Many different types of organizations are eligible to become Partners.

  1. Greening America's Capitals - Austin, TX

    Science.gov (United States)

    Report on the technical assistance project to help Austin, TX, develop a vision for the South Central Waterfront that incorporates green infrastructure to manage stormwater runoff, makes streets safer, and spurs investment.

  2. The Green Lacewing, Chrysoperla carnea

    DEFF Research Database (Denmark)

    Shrestha, Govinda; Enkegaard, Annie

    2013-01-01

    This study investigated the prey preference of 3rd instar green lacewing, Chrysoperla carnea Stephens (Neuroptera: Chrysopidae), between western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), and lettuce aphids, Nasonovia ribisnigri (Mosley) (Hemiptera: Aphididae...

  3. Green Construction in Building Renovation

    Science.gov (United States)

    Ksit, Barbara; Majcherek, Michał

    2016-06-01

    Modern materials and construction solutions draw more and more attention to ecology and building certification. Among the criteria appearing in revitalization, an important element is bringing plants back into heavily urbanized areas. In its natural form, this is not possible to carry out everywhere, often requiring large amounts of space. Nowadays, however, there are a number of green roofs and green wall systems, allowing "greener" construction without making significant changes in the urban environment. The article includes a presentation and analysis of selected solutions of biological surfaces known as green roofs and green walls, specifying various solutions and their most important features. The case study focuses primarily on material and design solutions, as well as the potential benefits, risks and limitations in their use. Plants structures on the surfaces of vertical and horizontal partitions continue to be a very interesting alternative to take into account when applying for grants, such as LEED or BREEAM certificates.

  4. Green corridors in freight logistics

    DEFF Research Database (Denmark)

    Panagakos, George

    of the performance of a green corridor in terms of pre-specified Key Performance Indicators (KPIs). The thesis builds on previous own work under the EUfinanced SuperGreen project and applies the new methodology on the GreCOR corridor extending from Oslo to Rotterdam. The scope of the two other objectives relates...... the rationale for a performance monitoring scheme has been established, the thesis critically reviews the SuperGreen methodology which consists of: (i) decomposing the corridor into transport chains, (ii) selecting a sample of typical chains, (iii) assessing these chains through a set of KPIs, and (iv......) aggregating the chain-level KPIs to corridor-level ones using proper weights. Unlike SuperGreen that suggests a study-based approach for constructing the corridor sample, the thesis proposes founding the selection of typical chains on the outcome of specialised transport models. The periodic collection...

  5. Promoting sustainability through green chemistry

    Energy Technology Data Exchange (ETDEWEB)

    Kirchhoff, Mary M. [American Chemical Society, 1155 Sixteenth Street, NW, Washington, DC 20036 (United States)

    2005-06-15

    Green chemistry is an important tool in achieving sustainability. The implementation of green chemistry, the design of chemical products and processes that reduce or eliminate the use and generation of hazardous substances, is essential if the expanding global population is to enjoy an increased standard of living without having a negative impact on the health of the planet. Cleaner technologies will allow the chemical enterprise to provide society with the goods and services on which it depends in an environmentally responsible manner. Green chemistry provides solutions to such global challenges as climate change, sustainable agriculture, energy, toxics in the environment, and the depletion of natural resources. A collaborative effort by industry, academia, and government is needed to promote the adoption of the green chemistry technologies necessary to achieve a sustainable society.

  6. Remote sensing: A green illusion

    Science.gov (United States)

    Soudani, Kamel; François, Christophe

    2014-02-01

    An analysis reveals that satellite-observed increases in canopy greenness during dry seasons, which were previously interpreted as positive responses of Amazon forests to more sunlight, are in fact an optical artefact. See Letter p.221

  7. Urban Green Infrastructure: German Experience

    OpenAIRE

    Diana Olegovna Dushkova; Sergey Nikolaevich Kirillov

    2016-01-01

    The paper presents a concept of urban green infrastructure and analyzes the features of its implementation in the urban development programmes of German cities. We analyzed the most shared articles devoted to the urban green infrastructure to see different approaches to definition of this term. It is based on materials of field research in the cities of Berlin and Leipzig in 2014-2015, international and national scientific publications. During the process of preparing the paper, consultations...

  8. GREEN MARKETING AND GLOBAL SCENARIOS‎

    OpenAIRE

    Dr. Jessy George; R. Jith

    2017-01-01

    Green Marketing must satisfy two objectives Improved Environmental Quality and Customer Satisfaction. Although no consumer product has a zero impact on the environment, in business, the terms "green product" and "environmental product" are used commonly to describe those that strive to protect or enhance the natural environment by conserving energy and/or resources and reducing or eliminating use of toxic agents, pollution, and waste. The present study has made an attempt to describe why the ...

  9. Green attitude and economic growth

    OpenAIRE

    Ott, Ingrid; Soretz, Susanne

    2015-01-01

    We analyse the interdependence between green attitude and equilibrium development of environmental quality in an endogenous growth model. Individuals take only part of their impact on pollution into account, hence there is a negative externality of capital accumulation on environmental quality. Increasing wealth or increasing pollution enhance green attitude and reduce the externality, because individuals care more about the environment if their income is higher or if pollution is more obviou...

  10. Green Construction in Building Renovation

    Directory of Open Access Journals (Sweden)

    Ksit Barbara

    2016-06-01

    Full Text Available Modern materials and construction solutions draw more and more attention to ecology and building certification. Among the criteria appearing in revitalization, an important element is bringing plants back into heavily urbanized areas. In its natural form, this is not possible to carry out everywhere, often requiring large amounts of space. Nowadays, however, there are a number of green roofs and green wall systems, allowing “greener” construction without making significant changes in the urban environment.

  11. Green Cloud on the Horizon

    Science.gov (United States)

    Ali, Mufajjul

    This paper proposes a Green Cloud model for mobile Cloud computing. The proposed model leverage on the current trend of IaaS (Infrastructure as a Service), PaaS (Platform as a Service) and SaaS (Software as a Service), and look at new paradigm called "Network as a Service" (NaaS). The Green Cloud model proposes various Telco's revenue generating streams and services with the CaaS (Cloud as a Service) for the near future.

  12. THE MEDIATION EFFECTS OF GREEN PERCEIVED CONSUMER SKEPTICISM AND GREEN PERCEIVED RISK IN THE RELATIONSHIP BETWEEN GREENWASHING AND GREEN TRUST

    OpenAIRE

    LEBLEBİCİ KOÇER, Leyla; DELİCE, Tuğba

    2017-01-01

    This study was carried out on the consumers in Kayseri and explores the influenceof greenwashing on green trust and discusses the mediation roles of green perceivedskepticism and green perceived risk. 430 consumers were participate in the research.Structural equation modelling was applied to test the research hypothesis. Thisstudy found that greenwashing has a negative association with green trust. In addition,it found that greenwashing is positively associated with green perceived consumersk...

  13. Managing consumer disengagement through green advertising strategies

    OpenAIRE

    Loh, E-Jian

    2017-01-01

    A discord is apparent between consumers’ concerns and their actual green behaviour – a surfacing issue that has obstructed effective green messages by advertisers. Four distinct consumer disengagement issues, including consumer backlash, environmental exhaustion, motivational challenges and social pressures were identified. This thesis explored cultural discourses surrounding green marketing and proposes green advertising strategies that address consumer disengagement. Using the Grounded Theo...

  14. Graham Greene: The Films of His Fiction.

    Science.gov (United States)

    Phillips, Gene D.

    This book makes a comparative study of the prose fiction of Graham Greene and the films made from that fiction. Special attention is focused upon the "cinematic" style of Greene's prose, the effect of Greene's screenwriting on his novels, and the characteristics of Greene's filmscripts. The book is divided into considerations of Greene…

  15. Determining the optimal number of individual samples to pool for quantification of average herd levels of antimicrobial resistance genes in Danish pig herds using high-throughput qPCR

    DEFF Research Database (Denmark)

    Clasen, Julie; Mellerup, Anders; Olsen, John Elmerdahl

    2016-01-01

    The primary objective of this study was to determine the minimum number of individual fecal samples to pool together in order to obtain a representative sample for herd level quantification of antimicrobial resistance (AMR) genes in a Danish pig herd, using a novel high-throughput qPCR assay...

  16. Evaluation by latent class analysis of a magnetic capture based DNA extraction followed by real-time qPCR as a new diagnostic method for detection of Echinococcus multilocularis in definitive hosts.

    Science.gov (United States)

    Maas, Miriam; van Roon, Annika; Dam-Deisz, Cecile; Opsteegh, Marieke; Massolo, Alessandro; Deksne, Gunita; Teunis, Peter; van der Giessen, Joke

    2016-10-30

    A new method, based on a magnetic capture based DNA extraction followed by qPCR, was developed for the detection of the zoonotic parasite Echinococcus multilocularis in definitive hosts. Latent class analysis was used to compare this new method with the currently used phenol-chloroform DNA extraction followed by single tube nested PCR. In total, 60 red foxes and coyotes from three different locations were tested with both molecular methods and the sedimentation and counting technique (SCT) or intestinal scraping technique (IST). Though based on a limited number of samples, it could be established that the magnetic capture based DNA extraction followed by qPCR showed similar sensitivity and specificity as the currently used phenol-chloroform DNA extraction followed by single tube nested PCR. All methods have a high specificity as shown by Bayesian latent class analysis. Both molecular assays have higher sensitivities than the combined SCT and IST, though the uncertainties in sensitivity estimates were wide for all assays tested. The magnetic capture based DNA extraction followed by qPCR has the advantage of not requiring hazardous chemicals like the phenol-chloroform DNA extraction followed by single tube nested PCR. This supports the replacement of the phenol-chloroform DNA extraction followed by single tube nested PCR by the magnetic capture based DNA extraction followed by qPCR for molecular detection of E. multilocularis in definitive hosts. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Green ergonomics: definition and scope.

    Science.gov (United States)

    Thatcher, Andrew

    2013-01-01

    This paper demonstrates that the goals of ergonomics (i.e. effectiveness, efficiency, health, safety and usability) are closely aligned with the goals of design for environmental sustainability. In this paper, the term 'green ergonomics' is conceptualised to specifically describe ergonomics interventions with a pro-nature emphasis. Green ergonomics is focused on the bi-directional connections between human systems and nature. This involves looking at (1) how ergonomics design and evaluation might be used to conserve, preserve, and restore nature and (2) how ecosystem services might be harnessed to facilitate the improved wellbeing and effectiveness of human systems. The paper proposes the scope of green ergonomics based on these bi-directional relationships in the areas of the design of low resource systems and products, the design of green jobs, and the design for behaviour change. Suggestions for further work in the green ergonomics domain are also made. Given the enormous environmental challenges facing modern industrial society, this paper encourages ergonomics science to embrace a pro-nature understanding of work design and research. This paper sets out the role for green ergonomics based on an appreciation of the human-nature connections that have been integrated with our understanding of ergonomics science and practice.

  18. Consumer perceptions of green power

    International Nuclear Information System (INIS)

    Rowlands, I.; Parker, P.; Scott, D.

    2001-01-01

    The relationship between consumer perceptions of the environmental impact of different energy sources and their willingness to pay a premium for green electricity was examined using the ANOVA analysis of variance and the chi-square test procedures. Since green power producers can include several energy resources in their offerings, it is important to understand the preferences of their customers. A survey sent to 480 residents in the Waterloo region of southern Ontario showed that when asked about purchasing nuclear, large scale hydropower or natural gas out of a choice of 11 energy sources, there was a wide discrepancy between those who stated a willingness to pay a large premium for green power and those who stated a willingness to pay only a small, or no, premium for green power. It was determined that these 3 energy resources were not popular among the most environmentally inclined portion of the consumer market. It was noted however that willingness to pay should be interpreted with caution because peoples' stated intentions to pay a premium for green power do not necessarily translate into action once they have the opportunity to buy power from renewable energy sources. However, marketing strategies could be used to improve the uptake of green power by consumers. The study suggests that landfill gas, is probably preferred to large-scale hydro. It was also suggested that the relative position of biomass, natural gas, garbage and nuclear power should be examined more closely. 29 refs., 3 tabs., 3 figs

  19. GREEN CHEMISTRY: NEW CHEMICAL PHILOSOPHY

    Directory of Open Access Journals (Sweden)

    F. A. Tykhomirova

    2015-11-01

    Full Text Available The review deals with the principles and guidelines of “Green chemistry” in comparison with the philosophy of nanotechnology. Modern philosophy and methodology of science research focus is on the process of the growth of scientific knowledge. Modern chemistry is complex, hierarchical, multilevel and multidimensional system. Philosophy of nanotechnology relies heavily on the value of scientism and the idea of domination of man over nature , there is an apology of human intervention in nature. “Green chemistry” is called “new thinking”of chemistry, philosophy of modern chemical research. The chemicals and processes in accordance with the principles of “Green chemistry” are considered not only in terms of production of substances and materials with desired properties, but also taking into account the consequences for the environment. In the “Green chemistry” created image of the “ideal customer” – he uses a minimum number of products understands the need to preserve the environment. Ideological landmark “Green chemistry” – co-evolution of man and nature, preservation of the biosphere. It emphasized the need to implement the ideology of “Green chemistry” in the training of future specialists.

  20. Brief Discussion on Green Building Materials

    International Nuclear Information System (INIS)

    Cai, Jia-wei; Sun, Jian

    2014-01-01

    With more and more emphasizes on the environment and resources, the concept of green buildings has been widely accepted. Building materials are vectors of architectures, only if green building materials and related technical means are used, can we construct green buildings to achieve the purpose of energy conservation and environmental protection. This paper introduces the relationship between green building materials and green buildings, the current situation of green building materials in China, as well as the measures to accelerate the development of green building materials

  1. A green hydrogen economy

    Energy Technology Data Exchange (ETDEWEB)

    Clark, W.W. II [Clark Communications, Beverly Hills, CA (United States). Green Hydrogen Scientific Advisory Committee; Rifkin, J. [The Foundation on Economic Trends (United States)

    2006-11-15

    This paper is the result of over a dozen scholars and practitioners who strongly felt that a hydrogen economy and hence the future is closer than some American politicians and bureaucrats state. Moreover, when seen internationally, there is strong evidence, the most recent and obvious ones are the proliferation of hybrid vehicles, that for any nation-state to be energy independent it must seek a renewable or green hydrogen future in the near term. The State of California has once again taken the lead in this effort for both an energy-independent future and one linked strongly to the hydrogen economy. Then why a hydrogen economy in the first instance? The fact is that hydrogen most likely will not be used for refueling of vehicles in the near term. The number of vehicles to make hydrogen commercially viable will not be in the mass market by almost all estimates until 2010. However, it is less than a decade away. The time frame is NOT 30-40 years as some argue. The hydrogen economy needs trained people, new ventures and public-private partnerships now. The paper points out how the concerns of today, including higher costs and technologies under development, can be turned into opportunities for both the public and private sectors. It was not too long ago that the size of a mobile phone was that of a briefcase, and then almost 10 years ago, the size of a shoe box. Today, they are not only the size of a man's wallet but also often given away free to consumers who subscribe or contract for wireless services. While hydrogen may not follow this technological commercialization exactly, it certainly will be on a parallel path. International events and local or regional security dictate that the time for a hydrogen must be close at hand. (author)

  2. A green hydrogen economy

    International Nuclear Information System (INIS)

    Clark, Woodrow W.; Rifkin, Jeremy

    2006-01-01

    This paper is the result of over a dozen scholars and practitioners who strongly felt that a hydrogen economy and hence the future is closer than some American politicians and bureaucrats state. Moreover, when seen internationally, there is strong evidence, the most recent and obvious ones are the proliferation of hybrid vehicles, that for any nation-state to be energy independent it must seek a renewable or green hydrogen future in the near term. The State of California has once again taken the lead in this effort for both an energy-independent future and one linked strongly to the hydrogen economy. Then why a hydrogen economy in the first instance? The fact is that hydrogen most likely will not be used for refueling of vehicles in the near term. The number of vehicles to make hydrogen commercially viable will not be in the mass market by almost all estimates until 2010. However, it is less than a decade away. The time frame is NOT 30-40 years as some argue. The hydrogen economy needs trained people, new ventures and public-private partnerships now. The paper points out how the concerns of today, including higher costs and technologies under development, can be turned into opportunities for both the public and private sectors. It was not too long ago that the size of a mobile phone was that of a briefcase, and then almost 10 years ago, the size of a shoe box. Today, they are not only the size of a man's wallet but also often given away free to consumers who subscribe or contract for wireless services. While hydrogen may not follow this technological commercialization exactly, it certainly will be on a parallel path. International events and local or regional security dictate that the time for a hydrogen must be close at hand

  3. Green power marketing

    Energy Technology Data Exchange (ETDEWEB)

    Wiltshire, S. [Selectpower Inc., Guelph, ON (Canada)

    2005-07-01

    Selectpower Inc. is an unregulated affiliate of Guelph Hydro and was formed to market green energy alternatives. Details of their Selectwind program were reviewed in this presentation. The program is available to both individuals and organizations. Customers sign a 3 or 5 year agreement to purchase monthly blocks of wind energy at a premium of $6.53 per month, which is billed on their Hydro bill. Details of the program's business strategy and branding policy were presented. The program markets itself by using full page colour newspaper ads, direct mailing and making forms available at Selectpower retail stores, mall kiosks and community events. In addition, Selectwind leaders are profiled in Enernews, and also have a quarterly newsletter. An example of an order form was provided, as well as an outline of Selectwind educational materials and details of their quality assurance procedures, EcoLogo certification and guarantees. Fifty percent of customers currently buy more than 100 kWh per month, and several customers buy 100 per cent equivalent of their electricity use as Selectwind. Minimum Selectwind purchase is 1200 kWh per year with a 3 year contract. Approximately 100 MWh are purchased every month, and 3,607,494 kWh have been sold for the life of the contracts. Selectwind's combined emissions reduction commitments are 3,206 tonnes of CO{sub 2}. It was observed that 45,000 Ontario customer using wind energy represented 6.2 MW of installed capacity. refs., tabs., figs.

  4. Green Power Initiative

    Energy Technology Data Exchange (ETDEWEB)

    Butler, Patrick Barry [Univ. of Iowa, Iowa City, IA (United States)

    2013-01-28

    National energy policy supports the gathering of more detailed and authoritative data on the introduction of renewable bio-based fuels into new and existing district energy systems via the application of biomass gasification. The University of Iowa developed a biomass-fueled, university-scale steam generation system based on biomass gasification technologies. The system serves as a state-of-the-art research and educational facility in the emerging application of gasification in steam generation. The facility, which includes a smaller down-draft gasifier and a larger multi-stage biomass boiler, was designed to operate primarily on wood-based fuels, but has provisions for testing other biomass fuel sources produced within a 100-mile radius, providing enough flexibility to meet the fluctuating local supply of biomass from industry and Midwest agriculture. The equipment was installed in an existing, staffed facility. The down-draft gasifier unit is operated by College of Engineering staff and students, under the direct technical supervision of qualified Utilities plant staff. The Green Power Initiative also includes a substantial, innovative educational component. In addition to an onsite, graduate-level research program in biomass fuels, the investigators have integrated undergraduate and graduate level teaching – through classroom studies and experiential learning – and applied research into a biomass-based, university-scale, functioning power plant. University of Iowa is unique in that it currently has multiple renewable energy technologies deployed, including significant biomass combustion (oat hulls) at its Main Power Plant and a new reciprocating engine based renewable district energy system. This project complements and supports the national energy policy and State of Iowa initiatives in ethanol and biodiesel. Byproducts of ethanol and biodiesel processes (distiller grains) as well as industry residues (oat hulls, wood chips, construction and demolition

  5. Detection of subgenomic mRNA of feline coronavirus by real-time polymerase chain reaction based on primer-probe energy transfer (P-sg-QPCR).

    Science.gov (United States)

    Hornyák, Akos; Bálint, Adám; Farsang, Attila; Balka, Gyula; Hakhverdyan, Mikhayil; Rasmussen, Thomas Bruun; Blomberg, Jonas; Belák, Sándor

    2012-05-01

    Feline infectious peritonitis is one of the most severe devastating diseases of the Felidae. Upon the appearance of clinical signs, a cure for the infected animal is impossible. Therefore rapid and proper diagnosis for both the presence of the causative agent, feline coronavirus (FCoV) and the manifestation of feline infectious peritonitis is of paramount importance. In the present work, a novel real-time RT-PCR method is described which is able to detect FCoV and to determine simultaneously the quantity of the viral RNA. The new assay combines the M gene subgenomic messenger RNA (sg-mRNA) detection and the quantitation of the genome copies of FCoV. In order to detect the broadest spectrum of potential FCoV variants and to achieve the most accurate results in the detection ability the new assay is applying the primer-probe energy transfer (PriProET) principle. This technology was chosen since PriProET is very robust to tolerate the nucleotide substitutions in the target area. Therefore, this technology provides a very broad-range system, which is able to detect simultaneously many variants of the virus(es) even if the target genomic regions show large scale of variations. The detection specificity of the new assay was proven by positive amplification from a set of nine different FCoV strains and negative from the tested non-coronaviral targets. Examination of faecal samples of healthy young cats, organ samples of perished animals, which suffered from feline infectious peritonitis, and cat leukocytes from uncertain clinical cases were also subjected to the assay. The sensitivity of the P-sg-QPCR method was high, since as few as 10 genome copies of FCoV were detected. The quantitative sg-mRNA detection method revealed more than 10-50,000 times increase of the M gene sg-mRNA in organ materials of feline infectious peritonitis cases, compared to those of the enteric FCoV variants present in the faeces of normal, healthy cats. These results indicate the applicability of

  6. Frugal Innovation and Green Business Models

    DEFF Research Database (Denmark)

    Andersen, Maj Munch

    2015-01-01

    The literature on ‘green business models’ is rapidly developing these years. This paper suggests that much existing work on green business models lacks a deeper theoretical understanding of eco-innovation and the green economy. The paper forwards an evolutionary economic perspective on green...... business models. This perspective departs in important ways from other approaches to green business models the implications of which are sought clarified and discussed in the paper. The paper argues for the need to link up green business model innovation to aggregate green economic change. The paper posits...... that the greening of the economy has reached such a stage of maturity where a generic ‘green business model’ is apparent. The paper points to eight characteristics of eco-innovation on the basis of which key changes to the business model are identified and schematised for the different stages of the green economic...

  7. The skeptical green consumer revisited: testing the relationship between green consumerism and skepticism toward advertising

    NARCIS (Netherlands)

    Matthes, J.; Wonneberger, A.

    2014-01-01

    This article revisits the widely believed notion of the skeptical green consumer, in other words, that green consumers tend to distrust green advertising. Study 1, a survey of U.S. consumers, found no positive relationship between green consumerism and general ad skepticism. However, green

  8. Real-Time PCR Quantification of Chloroplast DNA Supports DNA Barcoding of Plant Species.

    Science.gov (United States)

    Kikkawa, Hitomi S; Tsuge, Kouichiro; Sugita, Ritsuko

    2016-03-01

    Species identification from extracted DNA is sometimes needed for botanical samples. DNA quantification is required for an accurate and effective examination. If a quantitative assay provides unreliable estimates, a higher quantity of DNA than the estimated amount may be used in additional analyses to avoid failure to analyze samples from which extracting DNA is difficult. Compared with conventional methods, real-time quantitative PCR (qPCR) requires a low amount of DNA and enables quantification of dilute DNA solutions accurately. The aim of this study was to develop a qPCR assay for quantification of chloroplast DNA from taxonomically diverse plant species. An absolute quantification method was developed using primers targeting the ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL) gene using SYBR Green I-based qPCR. The calibration curve was generated using the PCR amplicon as the template. DNA extracts from representatives of 13 plant families common in Japan. This demonstrates that qPCR analysis is an effective method for quantification of DNA from plant samples. The results of qPCR assist in the decision-making will determine the success or failure of DNA analysis, indicating the possibility of optimization of the procedure for downstream reactions.

  9. Green growth in the Netherlands

    International Nuclear Information System (INIS)

    Balde, K.; Boelens, A.; Brinksma, E.; Edens, B.; Hiethaar, S.; Klein, P.; Schenau, S.

    2011-04-01

    In 2009 the Ministerial Council Meeting of the Organisation for Economic Co-operation and Development (OECD) committed itself to a green growth strategy. Such a strategy fosters economic growth and development while ensuring that natural resources can continue to provide the ecosystem services on which our well-being relies. It also endorses investment, competition and innovation which will underpin sustained growth and give rise to new economic opportunities. Green growth provides both a policy strategy for implementing this economic transformation and a monitoring framework with a proposed set of indicators. This report presents an overview of the state of green growth in the Netherlands. It should be regarded as a benchmark for a more thorough and comprehensive assessment of green growth in the future. It is based on the set of indicators proposed by the OECD in their intermediate report of February 2011. Data relevant to the Dutch situation are presented for twenty of these indicators, illustrating the observed trends. The indicators are grouped in four themes. For the first theme, environmental efficiency of production, on the whole the indicators show increased efficiency. However, indicators such as greenhouse gas intensity, energy efficiency and material intensity show only relative decoupling, which on its own is not enough to ensure green growth. In addition, the increase in environmental efficiency is partly explained by substitution of imports for domestic production, which is not conducive to green growth on a global scale: the efficiency gains in domestic production, for example, are offset by increases in foreign greenhouse gas emissions. Water use and agricultural nutrient surpluses are the only indicators where absolute decoupling has occurred. The second theme contains indicators regarding the natural assets base. This group of indicators provides a mixed picture. Natural gas reserves are decreasing and the overall level of threat to animal

  10. ADSORPTION MALACHITE GREEN ON NATURAL ZEOLITE

    OpenAIRE

    Eko Ariyanto

    2012-01-01

    A natural zeolite was employed as adsorbent for reducing of malachite green from aqueous solution. A batch system was applied to study the adsorption of malachite green in single system on natural zeolite. The adsorption studies indicate that malachite green in single component system follows the second-order kinetics and the adsorption is diffusion process with two stages for malachite green. Malachite green adsorption isotherm follows the Langmuir model.

  11. ADSORPTION MALACHITE GREEN ON NATURAL ZEOLITE

    Directory of Open Access Journals (Sweden)

    Eko Ariyanto

    2012-02-01

    Full Text Available A natural zeolite was employed as adsorbent for reducing of malachite green from aqueous solution. A batch system was applied to study the adsorption of malachite green in single system on natural zeolite. The adsorption studies indicate that malachite green in single component system follows the second-order kinetics and the adsorption is diffusion process with two stages for malachite green. Malachite green adsorption isotherm follows the Langmuir model.

  12. Progress report on Green Deals 2012; Voortgangsrapportage Green Deals 2012

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2012-10-15

    In the Dutch governmental coalition agreement the Green Deal approach was announced in the autumn of 2010. The focus of the Green Deals is for people and companies to develop sustainable initiatives that contribute to economic growth. This progress report provides an overview of the deals that this bottom-up approach has yielded. The report also provides information on the progress of the deals and the interim results of the approach and the individual deals. Also attention is paid to how the 131 Green Deals score on innovation and entrepreneurship [Dutch] In het regeerakkoord van het kabinet is in het najaar van 2010 de Green Deal-aanpak aangekondigd. Centraal in de aanpak staat dat mensen en bedrijven zoveel mogelijk ruimte krijgen voor eigen duurzame initiatieven die bijdragen aan economische groei. Deze voortgangsrapportage geeft een overzicht van de deals die deze bottom-up aanpak heeft opgeleverd. De rapportage informeert bovendien over de voortgang van de deals en over de tussentijdse resultaten van zowel de aanpak als de afzonderlijke deals. Ook wordt gekeken hoe de 131 Green Deals scoren op innovatief vermogen en ondernemerschap.

  13. Green Mines green energy : establishing productive land on mine tailings

    Energy Technology Data Exchange (ETDEWEB)

    Tisch, B.; Zinck, J.; Vigneault, B. [Natural Resources Canada, Ottawa, ON (Canada). CANMET Mining and Mineral Sciences Laboratories

    2009-02-15

    The Green Mines green energy research project was initiated by the CANMET Mining and Mineral Sciences Laboratories of Natural Resources Canada. The objective of the initiative was to demonstrate that organic residuals could be used to remediate mine tailings and establish agriculturally productive land where energy crops such as corn, canola, soy, switchgrass and other species could be grown and harvested specifically as feedstock for the production of green fuels. This paper discussed the scope and progress to date of the Green Mines green energy project. This included discussion about a column leaching study and about effluent treatability and toxicity. Neutralization test results and the results of field trials were presented. The paper concluded with a discussion of next steps. An advisory committee has been established to review annual progress and establish research directions. Overall, preliminary results from the column study suggest that sulphate reduction at the tailings-biosolids interface is occurring, although steady state has not yet been reached after more than one year of testing. 1 tab., 3 figs.

  14. Green Mines green energy : establishing productive land on mine tailings

    International Nuclear Information System (INIS)

    Tisch, B.; Zinck, J.; Vigneault, B.

    2009-01-01

    The Green Mines green energy research project was initiated by the CANMET Mining and Mineral Sciences Laboratories of Natural Resources Canada. The objective of the initiative was to demonstrate that organic residuals could be used to remediate mine tailings and establish agriculturally productive land where energy crops such as corn, canola, soy, switchgrass and other species could be grown and harvested specifically as feedstock for the production of green fuels. This paper discussed the scope and progress to date of the Green Mines green energy project. This included discussion about a column leaching study and about effluent treatability and toxicity. Neutralization test results and the results of field trials were presented. The paper concluded with a discussion of next steps. An advisory committee has been established to review annual progress and establish research directions. Overall, preliminary results from the column study suggest that sulphate reduction at the tailings-biosolids interface is occurring, although steady state has not yet been reached after more than one year of testing. 1 tab., 3 figs

  15. Making green infrastructure healthier infrastructure

    Directory of Open Access Journals (Sweden)

    Mare Lõhmus

    2015-11-01

    Full Text Available Increasing urban green and blue structure is often pointed out to be critical for sustainable development and climate change adaptation, which has led to the rapid expansion of greening activities in cities throughout the world. This process is likely to have a direct impact on the citizens’ quality of life and public health. However, alongside numerous benefits, green and blue infrastructure also has the potential to create unexpected, undesirable, side-effects for health. This paper considers several potential harmful public health effects that might result from increased urban biodiversity, urban bodies of water, and urban tree cover projects. It does so with the intent of improving awareness and motivating preventive measures when designing and initiating such projects. Although biodiversity has been found to be associated with physiological benefits for humans in several studies, efforts to increase the biodiversity of urban environments may also promote the introduction and survival of vector or host organisms for infectious pathogens with resulting spread of a variety of diseases. In addition, more green connectivity in urban areas may potentiate the role of rats and ticks in the spread of infectious diseases. Bodies of water and wetlands play a crucial role in the urban climate adaptation and mitigation process. However, they also provide habitats for mosquitoes and toxic algal blooms. Finally, increasing urban green space may also adversely affect citizens allergic to pollen. Increased awareness of the potential hazards of urban green and blue infrastructure should not be a reason to stop or scale back projects. Instead, incorporating public health awareness and interventions into urban planning at the earliest stages can help insure that green and blue infrastructure achieves full potential for health promotion.

  16. Making green infrastructure healthier infrastructure.

    Science.gov (United States)

    Lõhmus, Mare; Balbus, John

    2015-01-01

    Increasing urban green and blue structure is often pointed out to be critical for sustainable development and climate change adaptation, which has led to the rapid expansion of greening activities in cities throughout the world. This process is likely to have a direct impact on the citizens' quality of life and public health. However, alongside numerous benefits, green and blue infrastructure also has the potential to create unexpected, undesirable, side-effects for health. This paper considers several potential harmful public health effects that might result from increased urban biodiversity, urban bodies of water, and urban tree cover projects. It does so with the intent of improving awareness and motivating preventive measures when designing and initiating such projects. Although biodiversity has been found to be associated with physiological benefits for humans in several studies, efforts to increase the biodiversity of urban environments may also promote the introduction and survival of vector or host organisms for infectious pathogens with resulting spread of a variety of diseases. In addition, more green connectivity in urban areas may potentiate the role of rats and ticks in the spread of infectious diseases. Bodies of water and wetlands play a crucial role in the urban climate adaptation and mitigation process. However, they also provide habitats for mosquitoes and toxic algal blooms. Finally, increasing urban green space may also adversely affect citizens allergic to pollen. Increased awareness of the potential hazards of urban green and blue infrastructure should not be a reason to stop or scale back projects. Instead, incorporating public health awareness and interventions into urban planning at the earliest stages can help insure that green and blue infrastructure achieves full potential for health promotion.

  17. Microarray detection and qPCR screening of potential biomarkers of Folsomia candida (Collembola: Isotomidae) exposed to Bt proteins (Cry1Ab and Cry1Ac)

    DEFF Research Database (Denmark)

    Yuan, Yiyang; Krogh, Paul Henning; Bai, Xue

    2014-01-01

    The impact of Bt proteins on non-target arthropods is less understood than their effects on target organisms where the mechanism of toxic action is known. Here, we report the effects of two Bt proteins, Cry1Ab and Cry1Ac, on gene expression in the non-target collembolan, Folsomia candida....... A customized microarray was used to study gene expression in F. candida specimens that were exposed to Cry1Ab and Cry1Ac. All selected transcripts were subsequently confirmed by qPCR. Eleven transcripts were finally verified, and three of them were annotated. The responses of all eleven transcripts were...... tested in specimens for both Cry1Ab and Cry1Ac at a series of concentrations. These transcripts were separated into two and three groups for Cry1Ab and Cry1Ac, respectively, depend on their expression levels. However, those eleven transcripts did not respond to the Bt proteins in Bt-rice residues....

  18. Restriction Cascade Exponential Amplification (RCEA) assay with an attomolar detection limit: a novel, highly specific, isothermal alternative to qPCR.

    Science.gov (United States)

    Ghindilis, Andrey L; Smith, Maria W; Simon, Holly M; Seoudi, Ihab A; Yazvenko, Nina S; Murray, Iain A; Fu, Xiaoqing; Smith, Kenneth; Jen-Jacobson, Linda; Xu, Shuang-Yong

    2015-01-13

    An alternative to qPCR was developed for nucleic acid assays, involving signal rather than target amplification. The new technology, Restriction Cascade Exponential Amplification (RCEA), relies on specific cleavage of probe-target hybrids by restriction endonucleases (REase). Two mutant REases for amplification (Ramp), S17C BamHI and K249C EcoRI, were conjugated to oligonucleotides, and immobilized on a solid surface. The signal generation was based on: (i) hybridization of a target DNA to a Ramp-oligonucleotide probe conjugate, followed by (ii) specific cleavage of the probe-target hybrid using a non-immobilized recognition REase. The amount of Ramp released into solution upon cleavage was proportionate to the DNA target amount. Signal amplification was achieved through catalysis, by the free Ramp, of a restriction cascade containing additional oligonucleotide-conjugated Ramp and horseradish peroxidase (HRP). Colorimetric quantification of free HRP indicated that the RCEA achieved a detection limit of 10 aM (10(-17) M) target concentration, or approximately 200 molecules, comparable to the sensitivity of qPCR-based assays. The RCEA assay had high specificity, it was insensitive to non-specific binding, and detected target sequences in the presence of foreign DNA. RCEA is an inexpensive isothermal assay that allows coupling of the restriction cascade signal amplification with any DNA target of interest.

  19. Comparison of bacterial culture and qPCR testing of rectal and pen floor samples as diagnostic approaches to detect enterotoxic Escherichia coli in nursery pigs

    DEFF Research Database (Denmark)

    Weber, N. R.; Nielsen, J. P.; Hjulsager, Charlotte Kristiane

    2017-01-01

    Enterotoxigenic E. coli (ETEC) are a major cause of diarrhoea in weaned pigs. The objective of this study was to evaluate the agreement at pen level among three different diagnostic approaches for the detection of ETEC in groups of nursery pigs with diarrhoea. The diagnostic approaches used were...... to determine the quantity of F18 and F4 genes. The study was carried out in three Danish pig herds and included 31 pens with a pen-level diarrhoea prevalence of > 25%, as well as samples from 93 diarrhoeic nursery pigs from these pens. All E. coli isolates were analysed by PCR and classified as ETEC when genes...... was observed between the detection of ETEC by bacterial culture and qPCR in the same pen floor sample in 26 (83.9%, Kappa = 0.679) pens. Conclusion: We observed an acceptable agreement for the detection of ETEC-positive diarrhoeic nursery pigs in pen samples for both bacterial culture of pen floor samples...

  20. Chitinase mRNA Levels Determined by QPCR in Crab-Eating Monkey (Macaca fascicularis) Tissues: Species-Specific Expression of Acidic Mammalian Chitinase and Chitotriosidase.

    Science.gov (United States)

    Uehara, Maiko; Tabata, Eri; Ishii, Kazuhiro; Sawa, Akira; Ohno, Misa; Sakaguchi, Masayoshi; Matoska, Vaclav; Bauer, Peter O; Oyama, Fumitaka

    2018-05-09

    Mice and humans express two active chitinases: acidic mammalian chitinase (AMCase) and chitotriosidase (CHIT1). Both chitinases are thought to play important roles in specific pathophysiological conditions. The crab-eating monkey ( Macaca fascicularis ) is one of the most frequently used nonhuman primate models in basic and applied biomedical research. Here, we performed gene expression analysis of two chitinases in normal crab-eating monkey tissues by way of quantitative real-time polymerase chain reaction (qPCR) using a single standard DNA molecule. Levels of AMCase and CHIT1 messenger RNAs (mRNAs) were highest in the stomach and the lung, respectively, when compared to other tissues. Comparative gene expression analysis of mouse, monkey, and human using monkey⁻mouse⁻human hybrid standard DNA showed that the AMCase mRNA levels were exceptionally high in mouse and monkey stomachs while very low in the human stomach. As for the CHIT1 mRNA, we detected higher levels in the monkey lung when compared with those of mouse and human. The differences of mRNA expression between the species in the stomach tissues were basically reflecting the levels of the chitinolytic activities. These results indicate that gene expression of AMCase and CHIT1 differs between mammalian species and requiring special attention in handling data in chitinase-related studies in particular organisms.

  1. Citrus huanglongbing: validation of Real-Time PCR (qPCR for the detection of Candidatus Liberibacter asiaticus and Candidatus Liberibacter americanus in Colombia

    Directory of Open Access Journals (Sweden)

    Jorge Evelio Ángel

    2014-12-01

    Full Text Available Citrus huanglongbing (HLB is the most destructive citrus disease. Two of the three known HLB-associated Candidatus Liberibacter species were recently found to be present in the Americas. In this study, eggs, nymphs and adults of Diaphorina citri Kuwayama (Hemiptera: Liviidae and suspect citrus plant materials were collected in 25 municipalities in the departments of Cundinamarca, Santander, Valle del Cauca, Meta and Quindio (Colombia. The detection sensitivity, specificity and assay performance of the 16S rDNA-based real-time PCR (qPCR were validated for the field survey of the disease in Colombia. The validation confirmed the reliability and robustness of the real-time PCR method for the detection of HLB bacteria in host citrus plant tissues and the vector D. citri. The diagnosis was performed for Candidatus Liberibacter asiaticus (Ca. L. asiaticus and for Candidatus Liberibacter americanus (Ca. L. americanus on 168 citrus plant material samples and 239 insect samples. Neither Ca. L. asiaticus nor Ca. L. americanus were detected in the host plants or insects vector, confirming the absence of the disease in the citrus-producing areas of Colombia.

  2. 78 FR 56703 - Office of Federal High-Performance Green Buildings; Green Building Advisory Committee...

    Science.gov (United States)

    2013-09-13

    ... Federal High-Performance Green Buildings; Green Building Advisory Committee; Notification of Upcoming... Green Building Advisory Committee Meeting (the Committee) and the schedule for a series of conference..., Designated Federal Officer, [[Page 56704

  3. Green IT in Practice How One Company is Approaching the Greening of Its IT

    CERN Document Server

    Hird, Gary

    2010-01-01

    Green IT in Practice, Second edition provides guidance on how to implement a Green IT programme. It will help you to formulate a Green IT policy, curb demand for data storage capacity, and lower the electricity consumption of the datacentre.

  4. Green power: electricity's blue box

    International Nuclear Information System (INIS)

    Paterson, M.

    2002-01-01

    The concept of green power is taking hold, and the speaker briefly outlined the various options available at the moment, such as low impact hydro, wind, solar, and biomass. The electrical generation mix will be affected as a result of the provision of alternate power sources of energy to the community. It is possible for members of the community to make a well-informed choice as the eco-logo is the official certification of green products and power. Wind turbine interference with bird migration represents one of the main concerns attributed to wind power along with the generation of noise. The speaker indicated that the turbines are mostly located away from migration routes and that they turn very slowly, therefore not likely to cause a noise pollution problem for the public. The higher cost associated with green power causes a problem when compared with the cost or more traditional energy generation methods. Until they become more competitive, green power alternatives only fill a niche market. Ontario Power Generation is determined to assist the community with a move toward green energy in the future, especially given its position as an industry leader. refs., figs

  5. Greening the global water system

    Science.gov (United States)

    Hoff, H.; Falkenmark, M.; Gerten, D.; Gordon, L.; Karlberg, L.; Rockström, J.

    2010-04-01

    SummaryRecent developments of global models and data sets enable a new, spatially explicit and process-based assessment of green and blue water in food production and trade. An initial intercomparison of a range of different (hydrological, vegetation, crop, water resources and economic) models, confirms that green water use in global crop production is about 4-5 times greater than consumptive blue water use. Hence, the full green-to-blue spectrum of agricultural water management options needs to be used when tackling the increasing water gap in food production. The different models calculate considerable potentials for complementing the conventional approach of adding irrigation, with measures to increase water productivity, such as rainwater harvesting, supplementary irrigation, vapour shift and soil and nutrient management. Several models highlight Africa, in particular sub-Saharan Africa, as a key region for improving water productivity in agriculture, by implementing these measures. Virtual water trade, mostly based on green water, helps to close the water gap in a number of countries. It is likely to become even more important in the future, when inequities in water availability are projected to grow, due to climate, population and other drivers of change. Further model developments and a rigorous green-blue water model intercomparison are proposed, to improve simulations at global and regional scale and to enable tradeoff analyses for the different adaptation options.

  6. Green functions of vortex operators

    International Nuclear Information System (INIS)

    Polchinski, J.; California Univ., Berkeley

    1981-01-01

    We study the euclidean Green functions of the 't Hooft vortex operator, primarly for abelian gauge theories. The operator is written in terms of elementary fields, with emphasis on a form in which it appears as the exponential of a surface integral. We explore the requirement that the Green functions depend only on the boundary of this surface. The Dirac veto problem appears in a new guise. We present a two-dimensional solvable model of a Dirac string, which suggests a new solution of the veto problem. The renormalization of the Green functions of the abelian Wilson loop and abelian vortex operator is studied with the aid of the operator product expansion. In each case, an overall multiplication of the operator makes all Green functions finite; a surprising cancellation of divergences occurs with the vortex operator. We present a brief discussion of the relation between the nature of the vacuum and the cluster properties of the Green functions of the Wilson and vortex operators, for a general gauge theory. The surface-like cluster property of the vortex operator in an abelian Higgs theory is explored in more detail. (orig.)

  7. Green commercial building insurance in Malaysia

    Science.gov (United States)

    Yang, Yu Xin Ou; Chew, Boon Cheong; Loo, Heoy Shin; Tan, Lay Hong

    2017-03-01

    Green building construction is growing tremendously globally even in Malaysia. Currently, there are approximate 636 buildings have registered and to be certified with Green Building Index. Among these buildings, 45 buildings have already fulfilled the requirements and fully certified. The other buildings still under provisional certification stage. Malaysia had adopted Green Building Index in 2009 to support a move to promote green building concept. Malaysia starts to move towards green building because Malaysian construction and building industry realizes that both energy consumed and waste produced are reduced without irreversible impacts to ecosystems. Consequently, insurance companies such as Fireman's Fund from America has started the green building insurance policies for their green building in the year of 2006, while Malaysia still remain the coverage for green buildings using conventional property insurance. There are lacks of efforts to be seen from insurance companies to propose green building insurance for these green buildings. There are a few factors which can take into consideration for insurance companies to start the very first green building insurance in Malaysia. Although there are challenges, some efficient strategies have been identified to overcome the problems. The methods used in this research topic is qualitative research. The results obtained shows that green commercial building insurance has a huge business opportunity in Malaysia because the number of green commercial buildings are increasing tremendously in Malaysia. It is a favor to implement green building insurance in Malaysia. Furthermore, insurance companies can consider to add in extra coverage in standard building policy to provide extra protection for non-certified green buildings which have the intention to rebuilt in green when damage happens. Generally, it is very important to introduce green commercial buildings insurance into Malaysia so that all of the green commercial

  8. Green justice in the city

    DEFF Research Database (Denmark)

    Rutt, Rebecca Leigh; Gulsrud, Natalie Marie

    2016-01-01

    tIn this short communication, we discuss European urban green space (UGS) research from an environ-mental justice perspective. We show that European UGS scholarship primarily focuses on functionalvalues and managerial aspects of UGS, while paying less attention to equity in the enjoyment of andde...... of anddecision-making around UGS. On this basis we discuss potentials for European urban green space researchto take up a more explicit environmental justice framing to shed much-needed light on injustices inEuropean cities and inspire change in policy and practice.......tIn this short communication, we discuss European urban green space (UGS) research from an environ-mental justice perspective. We show that European UGS scholarship primarily focuses on functionalvalues and managerial aspects of UGS, while paying less attention to equity in the enjoyment...

  9. Managing Green Business Model Transformations

    CERN Document Server

    Sommer, Axel

    2012-01-01

    Environmental sustainability creates both tremendous business opportunities and formidable threats to established companies across virtually all industry sectors. Yet many companies tackle the issue in a superficial or passive way through increased environmental reporting, the use of “greenspeak” in their corporate communication activities or isolated efforts to create green products or reduce pollution. In contrast, there are a small but increasing number of firms that employ a holistic approach to sustainability and consider fundamental changes to their existing business models. By ignoring the opportunities of Green Business Model Transformations, companies exclude themselves from a large variety of potential means to create economic value. In addition to ordinary product and process innovations, they can change “the rules of the game” within an industry towards environmental sustainability. This can facilitate the commercialisation of new green products that would not be competitive otherwise targ...

  10. Green roofs : a watertight perspective

    Energy Technology Data Exchange (ETDEWEB)

    Honza, D. [Honza Group Inc., Columbia, MD (United States)]|[Barrett Co., Millington, NJ (United States)

    2005-07-01

    While there is a growing acceptance of the ecological benefits of green roofs, many roofing contractors view green roofs with suspicion. The roofing industry is a high-volume, low-margin cost-driven industry which promotes a minimum standard commodity mentality. Roofing and waterproofing is the largest source of claims against architects and engineers. This paper suggested that architectural firms and engineering firms can reduce many issues associated with roofing problems by investigating and understanding materials, demands of projects, and preparing thorough specifications. Long-term exposure to the sun's rays will impact the chemical make-up of the roofing material, and water can break down the surface molecular structure of the membrane. Daily, yearly and event-related temperature variations can subject membranes to thermal induced stresses. Many roofs leak as a result of abuse during construction. Understanding and anticipating the performance problems of membranes can give green roof designers the ability to address limitations through good design. The membrane for a green roof should have superior abuse resistance; elastic properties, and resistance to long-term wet or saturated environments. Flashings for green roofs must exceed minimum standards. Membranes should be tested for watertightness before components are installed using electronic field vector mapping. Overburden should be installed after the membrane installation is proven to be watertight. It was concluded that higher design standards are required for green roofs, as many traditional roof membranes fail prematurely. A review of widely used membranes in the roofing and waterproofing industry included modified bitumen; built-up roofing; cured synthetic rubber sheets; thermalplastic membranes; self-adhering modified bitumen; and rubberized asphalt. 6 refs., 2 tabs., 6 figs.

  11. Growing a green economy in China

    Science.gov (United States)

    Weng, Qingqing; Xu, He; Ji, Yijun

    2018-02-01

    With the rapid development of economy, resource depletion and environmental degradation have become serious challenges for Chinese sustainable development. Green development is a mode of well environmental and high-quality economic development. It is necessary for China to implement green development. In this review, it discusses the green development problems in China, the international experience and connotation of green development are summarized and identified further. Based on the connotation and experience of green economy development, it puts forward several countermeasures and suggestions for Chinese green development finally.

  12. SISH/CISH or qPCR as alternative techniques to FISH for determination of HER2 amplification status on breast tumors core needle biopsies: a multicenter experience based on 840 cases.

    Science.gov (United States)

    Jacquemier, Jocelyne; Spyratos, Frédérique; Esterni, Benjamin; Mozziconacci, Marie-Joëlle; Antoine, Martine; Arnould, Laurent; Lizard, Sarab; Bertheau, Philippe; Lehmann-Che, Jacqueline; Fournier, Cécile Blanc; Krieger, Sophie; Bibeau, Frédéric; Lamy, Pierre-Jean; Chenard, Marie Pierre; Legrain, Michèle; Guinebretière, Jean-Marc; Loussouarn, Delphine; Macgrogan, Gaëtan; Hostein, Isabelle; Mathieu, Marie Christine; Lacroix, Ludovic; Valent, Alexander; Robin, Yves Marie; Revillion, Françoise; Triki, Magali Lacroix; Seaume, Aline; Salomon, Anne Vincent; de Cremoux, Patricia; Portefaix, Geneviève; Xerri, Luc; Vacher, Sophie; Bièche, Ivan; Penault-Llorca, Frédérique

    2013-07-22

    Until now, FISH has been the gold standard technique to identify HER2 amplification status in ambiguous cases of breast cancer. Alternative techniques have been developed to increase the capacities of investigating HER2 amplification status. The aims of this multicenter study in a large series of breast cancer patients were to prospectively compare the level of performance of CISH, SISH, and qPCR alternative techniques on paraffin-embedded core biopsies with "gold standard FISH" for evaluation of HER2 amplification status. This study was performed on 840 cases scored by immunohistochemistry (IHC): 0=317 (38%), 1+=183 (22%), 2+=109 (13%), 3+=231 (27%). Each of the 15 French centers participating in the study analyzed 56 breast carcinoma cases diagnosed on fixed paraffin-embedded core biopsies. HER2 amplification status was determined by commercially available FISH used as the reference technique with determination of the HER2/CEN17 ratio or HER2 copy number status. The alternative techniques performed on the same cases were commercially available SISH or CISH and a common qPCR method especially designed for the study including a set of 10 primer pairs: 2 for HER2 (exons 8 and 26), 5 to evaluate chromosome 17 polysomy TAOK1, UTP6, MRM1, MKS1, SSTR2 and 3 for diploidy control TSN, LAP3 and ADAMTS16. The concordance between IHC and FISH was 96% to 95% based on the HER2/CEN17 ratio (n=766) or HER2 copy number (n=840), respectively. The concordance of the alternative techniques with FISH was excellent: 97% and 98% for SISH (498 and 587 cases), 98% and 75% for CISH (108 and 204 cases) and 95% and 93% (699 and 773 cases) for qPCR based on the HER2/CEN17 ratio or HER2 copy number, respectively. Similarly, sensitivity ranged from 99% to 95% for SISH, 100% to 99% for CISH and 89% to 80% for qPCR. The concordance with FISH (ratio) in the 2+ cases was 89% for SISH, 100% for CISH and 93% for qPCR. These alternative techniques showed an excellent concordance with FISH in core

  13. Green's Theorem for Sign Data

    OpenAIRE

    Houston, Louis M.

    2012-01-01

    Sign data are the signs of signal added to noise. It is well known that a constant signal can be recovered from sign data. In this paper, we show that an integral over variant signal can be recovered from an integral over sign data based on the variant signal. We refer to this as a generalized sign data average. We use this result to derive a Green's theorem for sign data. Green's theorem is important to various seismic processing methods, including seismic migration. Results in this paper ge...

  14. The making of green knowledge

    DEFF Research Database (Denmark)

    Jamison, Andrew

    2003-01-01

    of such "green knowledge". The general argument is that, in the future, new types of interaction and new spaces for communication will need to be developed if green knowledge is not to be incorporated into the dominant culture or reduced to ineffective forms of protest.......The paper discusses some of the contributions of environmental activism to the development of knowledge. The paper contrasts some of the main forms of knowledge-making that have emerged among activists and raises a number of questions both about the political and cognitive implications...

  15. Greening and “un”greening Adelaide, South Australia

    Directory of Open Access Journals (Sweden)

    Guy M. Robinson

    2015-06-01

    Full Text Available The original design for Adelaide, the capital city of the state of South Australia, incorporated a green belt (known as the Park Lands around the city centre, itself laid out on a one square mile (2.59 km2 grid and including five large public squares. The Park Lands provided a barrier to urban sprawl and covered approximately 9.31 km2, of which 1.53 km2 has been used subsequently for cultural institutions, railways, cemeteries, sporting facilities and other constructions. In addressing issues of greening pertaining to Adelaide, the Park Lands and its management represents a core element in the evolving history of the city's growth. This paper will consider some of the contradictions within this growth, examining the changing attitudes of government and the populace to the Park Lands and also to the increasing sprawl of the city. It can be argued that this sprawl has been antithetical to maintenance of biodiversity and principles of “greening”, not only during the main phase of expansion in the 1960s and 1970s but also in recent years when planned development on prime farmland and other “green” areas is contributing to problems for provision of transport infrastructure and generally reducing capacity for sustainability. The potential for conflict between the desire to maintain biodiversity versus protection for the growing number of people moving into bushfire risk areas is just one of several examples of problems arising as a result of a relaxed attitude to low-density expansion. In examining these problems the paper will present maps of the changing footprint of Adelaide and will elaborate new “greening” initiatives that include green roofs, new systems of water harvesting, community-supported agriculture and schemes directly aimed at creating low-carbon living. A consistent theme will be the contradictions within plans for the city between greening and “un”greening.

  16. Fungal Microbiomes Associated with Green and Non-Green Building Materials.

    Science.gov (United States)

    Coombs, Kanistha; Vesper, Stephen; Green, Brett J; Yermakov, Mikhail; Reponen, Tiina

    2017-01-01

    Water-damaged buildings can lead to fungal growth and occupant health problems. Green building materials, derived from renewable sources, are increasingly utilized in construction and renovations. However, the question as to what fungi will grow on these green compared to non-green materials, after they get wet, has not been adequately studied. By determining what fungi grow on each type of material, the potential health risks can be more adequately assessed. In this study, we inoculated green and non-green pieces of ceiling tile, composite board, drywall, and flooring with indoor dust containing a complex mixture of naturally occurring fungi. The materials were saturated with water and incubated for two months in a controlled environment. The resulting fungal microbiomes were evaluated using ITS amplicon sequencing. Overall, the richness and diversity of the mycobiomes on each pair of green and non-green pieces were not significantly different. However, different genera dominated on each type of material. For example, Aspergillus spp. had the highest relative abundance on green and non-green ceiling tiles and green composite boards, but Peniophora spp. dominated the non-green composite board. In contrast, Penicillium spp. dominated green and non-green flooring samples. Green gypsum board was dominated by Phialophora spp. and Stachybotrys spp., but non-green gypsum board by Myrothecium spp. These data suggest that water-damaged green and non-green building materials can result in mycobiomes that are dominated by fungal genera whose member species pose different potentials for health risks.

  17. Detection of Angiostrongylus cantonensis in the Blood and Peripheral Tissues of Wild Hawaiian Rats (Rattus rattus by a Quantitative PCR (qPCR Assay.

    Directory of Open Access Journals (Sweden)

    Susan I Jarvi

    Full Text Available The nematode Angiostrongylus cantonensis is a rat lungworm, a zoonotic pathogen that causes human eosinophilic meningitis and ocular angiostrongyliasis characteristic of rat lungworm (RLW disease. Definitive diagnosis is made by finding and identifying A. cantonensis larvae in the cerebral spinal fluid or by using a custom immunological or molecular test. This study was conducted to determine if genomic DNA from A. cantonensis is detectable by qPCR in the blood or tissues of experimentally infected rats. F1 offspring from wild rats were subjected to experimental infection with RLW larvae isolated from slugs, then blood or tissue samples were collected over multiple time points. Blood samples were collected from 21 rats throughout the course of two trials (15 rats in Trial I, and 6 rats in Trial II. In addition to a control group, each trial had two treatment groups: the rats in the low dose (LD group were infected by approximately 10 larvae and the rats in the high dose (HD group were infected with approximately 50 larvae. In Trial I, parasite DNA was detected in cardiac bleed samples from five of five LD rats and five of five HD rats at six weeks post-infection (PI, and three of five LD rats and five of five HD rats from tail tissue. In Trial II, parasite DNA was detected in peripheral blood samples from one of two HD rats at 53 minutes PI, one of two LD rats at 1.5 hours PI, one of two HD rats at 18 hours PI, one of two LD rats at five weeks PI and two of two at six weeks PI, and two of two HD rats at weeks five and six PI. These data demonstrate that parasite DNA can be detected in peripheral blood at various time points throughout RLW infection in rats.

  18. Detection of Angiostrongylus cantonensis in the Blood and Peripheral Tissues of Wild Hawaiian Rats (Rattus rattus) by a Quantitative PCR (qPCR) Assay.

    Science.gov (United States)

    Jarvi, Susan I; Pitt, William C; Farias, Margaret E; Shiels, Laura; Severino, Michael G; Howe, Kathleen M; Jacquier, Steven H; Shiels, Aaron B; Amano, Karis K; Luiz, Blaine C; Maher, Daisy E; Allison, Maureen L; Holtquist, Zachariah C; Scheibelhut, Neil T

    2015-01-01

    The nematode Angiostrongylus cantonensis is a rat lungworm, a zoonotic pathogen that causes human eosinophilic meningitis and ocular angiostrongyliasis characteristic of rat lungworm (RLW) disease. Definitive diagnosis is made by finding and identifying A. cantonensis larvae in the cerebral spinal fluid or by using a custom immunological or molecular test. This study was conducted to determine if genomic DNA from A. cantonensis is detectable by qPCR in the blood or tissues of experimentally infected rats. F1 offspring from wild rats were subjected to experimental infection with RLW larvae isolated from slugs, then blood or tissue samples were collected over multiple time points. Blood samples were collected from 21 rats throughout the course of two trials (15 rats in Trial I, and 6 rats in Trial II). In addition to a control group, each trial had two treatment groups: the rats in the low dose (LD) group were infected by approximately 10 larvae and the rats in the high dose (HD) group were infected with approximately 50 larvae. In Trial I, parasite DNA was detected in cardiac bleed samples from five of five LD rats and five of five HD rats at six weeks post-infection (PI), and three of five LD rats and five of five HD rats from tail tissue. In Trial II, parasite DNA was detected in peripheral blood samples from one of two HD rats at 53 minutes PI, one of two LD rats at 1.5 hours PI, one of two HD rats at 18 hours PI, one of two LD rats at five weeks PI and two of two at six weeks PI, and two of two HD rats at weeks five and six PI. These data demonstrate that parasite DNA can be detected in peripheral blood at various time points throughout RLW infection in rats.

  19. Green Infrastructure Models and Tools

    Science.gov (United States)

    The objective of this project is to modify and refine existing models and develop new tools to support decision making for the complete green infrastructure (GI) project lifecycle, including the planning and implementation of stormwater control in urban and agricultural settings,...

  20. Green Fire and Religious Spirit.

    Science.gov (United States)

    Haluza-Delay, Randolph

    2000-01-01

    Spirituality, even within traditional religious practices, can inspire environmentally aware lifestyles. To help this "green fire" burn well, experiential educators should acknowledge religious beliefs and practices and avoid perpetuating the modern spiritual-secular rift. A "critical experientialism" is needed when it comes to…

  1. Co-Creating Green Transition

    DEFF Research Database (Denmark)

    Horsbøl, Anders

    2018-01-01

    Citizen involvement plays an important role in many governmental and municipal attempts towards green transition, reflecting a departure from a deficit model of public communication towards participatory ambitions of engaging citizens. Recently, the notion of co-creation or co-production has gained...

  2. Welcome to The Green Village

    NARCIS (Netherlands)

    Van Wijk, A.J.M.

    2013-01-01

    A sustainable world can only be achieved by an open collaboration between science, business and the public. That is why we create the Green Village: an innovative, lively, interactive and challenging environment where entrepreneurs, innovators, companies, artists, teachers and visitors can meet,

  3. Green Accounts of Christianshavn (VB)

    DEFF Research Database (Denmark)

    Østerby, Peder; Pedersen, Bjørn; Jørgensen, Michael Søgaard

    1999-01-01

    The project was to develope a Green Account of 1998. It is based on public data, the questionnaries filled out by 300 residents and some interviewes.The final account of '98 included 10 subjects which follows up on the '97 account. Besides that it presents two new subjects - the marked...

  4. Green Application for Space Power

    Science.gov (United States)

    Robinson, Joel

    2015-01-01

    Most space vehicle auxiliary power units (APUs) use hydrazine propellant for generating power. Hydrazine is a toxic, hazardous fuel that requires special safety equipment and processes for handling and loading. In recent years, there has been development of two green propellants (less toxic) that could enable their use in APUs. The Swedish government, in concert with the Swedish Space Corporation, has developed a propellant based on ammonium dinitramide (LMP-103S) that was flown on the Prisma spacecraft in 2010. The United States Air Force (USAF) has been developing a propellant based on hydroxylammonium nitrate (AFM315E) that is scheduled to fly on the Green Propellant Infusion Mission in the spring of 2016 to demonstrate apogee and reaction control thrusters. However, no one else in the Agency is currently pursuing use of green propellants for application to the APUs. Per the TA-01 Launch Propulsion Roadmap, the Space Technology Mission Directorate had identified the need to have a green propellant APU by 2015. This is our motivation for continuing activities.

  5. What Makes Green Schools Better?

    Science.gov (United States)

    Schimmel, Barry

    2011-01-01

    Green energy represents a way to empower students by demonstrating creative problem-solving with an eye on protecting precious resources, both capital and natural. Many school districts have already taken the initiative during the past five years to implement energy projects, whether for the educational or economic opportunities, or both.…

  6. Green Chemistry with Microwave Energy

    Science.gov (United States)

    Green chemistry utilizes a set of 12 principles that reduces or eliminates the use or generation of hazardous substances in the design, manufacture, and applications of chemical products (1). This newer chemical approach protects the environment by inventing safer and eco-friendl...

  7. Green Grows The Bottom Line

    Institute of Scientific and Technical Information of China (English)

    MARKGODFREY

    2004-01-01

    In promoting sustainable business practices in China, business leaders from the EU and Chinese companies have much to share but not that much in common. The EU is funding China's largest environmental program but, worried about the cost factor, some Chinese businesses areproving reluctant about going green.

  8. Green chemistry for chemical synthesis

    OpenAIRE

    Li, Chao-Jun; Trost, Barry M.

    2008-01-01

    Green chemistry for chemical synthesis addresses our future challenges in working with chemical processes and products by inventing novel reactions that can maximize the desired products and minimize by-products, designing new synthetic schemes and apparati that can simplify operations in chemical productions, and seeking greener solvents that are inherently environmentally and ecologically benign.

  9. Green chemistry for chemical synthesis.

    Science.gov (United States)

    Li, Chao-Jun; Trost, Barry M

    2008-09-09

    Green chemistry for chemical synthesis addresses our future challenges in working with chemical processes and products by inventing novel reactions that can maximize the desired products and minimize by-products, designing new synthetic schemes and apparati that can simplify operations in chemical productions, and seeking greener solvents that are inherently environmentally and ecologically benign.

  10. Educational benefits of green chemistry

    Science.gov (United States)

    Desmond, Serenity; Ray, Christian; Andino Martínez, José G.

    2017-08-01

    In this article, we present our current state of affairs in the "greening" of general chemistry laboratories, at the University of Illinois at Urbana-Champaign. We recognize the need to quantify our environmental mark and what we plan to do to continue to strive to make our work more sustainable and educational.

  11. Green corridors and network design

    DEFF Research Database (Denmark)

    Panagakos, George

    2016-01-01

    The purpose of this chapter is to investigate the relation between the Trans-European Transport Network (TEN-T) and the green corridor concept. First, the need is established for a corridor governance structure that enables the close cooperation among the numerous stakeholders from both the public...

  12. From green investments to ecotax

    International Nuclear Information System (INIS)

    Stam, B.

    1996-01-01

    Since 1991 several fiscal regulations are in effect for businesses, energy companies and civilians to invest in environmental and renewable projects. Although those so-called 'green investment' options are a great success, there is another, less positive, side to that success. Characteristics, advantages and disadvantages of several fiscal tools to invest in renewable energy are briefly discussed

  13. Growing your green chemistry mindset

    Science.gov (United States)

    Kosmas, Steven

    2017-08-01

    The purpose of this article is not to delineate the steps to move across the continuum to being a greener chemist, but to analyse the cognitive processes involved in fostering a green chemistry growth mindset (GCGM) [Dweck C. (2006) Mindset: The New Psychology of Success. New York, NY: Ballatine]. The focus is on changing the mindset, which inevitably will lead to a more mindful approach to chemistry practices before the laboratory begins. A green chemistry fixed mindset (GCFM) is closed to making improvements, since the attitude is that the techniques and processes in the laboratory are already employing a green chemistry mindset [Dweck C. (2006) Mindset: The New Psychology of Success. New York, NY: Ballatine]. The problem with the GCFM is that it precludes the possibility of making improvements. However, the GCGM employs a continuous, intentional focus on the attitude towards green chemistry, with the ultimate goal being a change in chemistry practices that is greener. The focus of this article will be on the GCGM.

  14. Green tribology: principles, research areas and challenges.

    Science.gov (United States)

    Nosonovsky, Michael; Bhushan, Bharat

    2010-10-28

    In this introductory paper for the Theme Issue on green tribology, we discuss the concept of green tribology and its relation to other areas of tribology as well as other 'green' disciplines, namely, green engineering and green chemistry. We formulate the 12 principles of green tribology: the minimization of (i) friction and (ii) wear, (iii) the reduction or complete elimination of lubrication, including self-lubrication, (iv) natural and (v) biodegradable lubrication, (vi) using sustainable chemistry and engineering principles, (vii) biomimetic approaches, (viii) surface texturing, (ix) environmental implications of coatings, (x) real-time monitoring, (xi) design for degradation, and (xii) sustainable energy applications. We further define three areas of green tribology: (i) biomimetics for tribological applications, (ii) environment-friendly lubrication, and (iii) the tribology of renewable-energy application. The integration of these areas remains a primary challenge for this novel area of research. We also discuss the challenges of green tribology and future directions of research.

  15. Planning multifunctional green infrastructure for compact cities

    DEFF Research Database (Denmark)

    Hansen, Rieke; Olafsson, Anton Stahl; van der Jagt, Alexander P.N.

    2018-01-01

    green space functions or the purposive design and management of multifunctional parks. Based on the findings, we arrive at five recommendations for promoting multifunctional urban green infrastructure in densifying urban areas: 1) undertake systematic spatial assessments of all urban green (and blue....... Further, spatial assessment, strategic planning and site design need to 4) consider synergies, trade-offs and the capacity of urban green spaces to provide functions as part of the wider green infrastructure network; and 5) largely benefit from cooperation between different sectors and public departments......Urban green infrastructure planning aims to develop green space networks on limited space in compact cities. Multifunctionality is considered key to achieving this goal as it supports planning practice that considers the ability of green spaces to provide multiple benefits concurrently. However...

  16. Green noise wall construction and evaluation.

    Science.gov (United States)

    2011-09-01

    This report details the research performed under Phase I of a research study titled Green Noise Wall Construction and Evaluation that looks into the feasibility of using green noise barriers as a noise mitigation option in Ohio. This phase incl...

  17. Green Power Partnership Top 30 Retail

    Science.gov (United States)

    EPA's Green Power Partnership is a voluntary program designed to reduce the environmental impact of electricity generation by promoting renewable energy. This list represents the largest green power users among retail partners within the GPP.

  18. Modeling for Green Supply Chain Evaluation

    Directory of Open Access Journals (Sweden)

    Elham Falatoonitoosi

    2013-01-01

    Full Text Available Green supply chain management (GSCM has become a practical approach to develop environmental performance. Under strict regulations and stakeholder pressures, enterprises need to enhance and improve GSCM practices, which are influenced by both traditional and green factors. This study developed a causal evaluation model to guide selection of qualified suppliers by prioritizing various criteria and mapping causal relationships to find effective criteria to improve green supply chain. The aim of the case study was to model and examine the influential and important main GSCM practices, namely, green logistics, organizational performance, green organizational activities, environmental protection, and green supplier evaluation. In the case study, decision-making trial and evaluation laboratory technique is applied to test the developed model. The result of the case study shows only “green supplier evaluation” and “green organizational activities” criteria of the model are in the cause group and the other criteria are in the effect group.

  19. Sizing up arthropod genomes: an evaluation of the impact of environmental variation on genome size estimates by flow cytometry and the use of qPCR as a method of estimation.

    Science.gov (United States)

    Gregory, T Ryan; Nathwani, Paula; Bonnett, Tiffany R; Huber, Dezene P W

    2013-09-01

    A study was undertaken to evaluate both a pre-existing method and a newly proposed approach for the estimation of nuclear genome sizes in arthropods. First, concerns regarding the reliability of the well-established method of flow cytometry relating to impacts of rearing conditions on genome size estimates were examined. Contrary to previous reports, a more carefully controlled test found negligible environmental effects on genome size estimates in the fly Drosophila melanogaster. Second, a more recently touted method based on quantitative real-time PCR (qPCR) was examined in terms of ease of use, efficiency, and (most importantly) accuracy using four test species: the flies Drosophila melanogaster and Musca domestica and the beetles Tribolium castaneum and Dendroctonus ponderosa. The results of this analysis demonstrated that qPCR has the tendency to produce substantially different genome size estimates from other established techniques while also being far less efficient than existing methods.

  20. Green accounting for greener energy

    International Nuclear Information System (INIS)

    Stanojevic, M.; Vranes, S.; Goekalp, I.

    2010-01-01

    The first step towards the widespread use of renewable energy sources and preservation of our environment for the people of the future is to adopt the ''green accounting'' standards that translate socially and environmentally responsible behavior into monetary terms, the only language businesses understand. These standards have the potential of switching on the red light for all pollution-causing power plants, and those depleting the natural capital in any way, be it over-harvesting the forests, or exhausting the underground treasures - coal, oil, natural gas, etc. This paper will show how green accounting can help in changing the focus from the economic welfare to the total societal welfare, acknowledging the fact that human society is an integral part of the natural world. The paper will also briefly present the software developed by the authors that introduce the green accounting principles into the investment appraisal process, aiming at encouraging investments into renewable energy. The tool is also useful as a platform facilitating calibration of economic/financial instruments, like environmental taxes of governmental incentives, that are usually to boost renewable energy sector. The comparative analysis of investment into biofuel-powered combined heat and power production plant using two types of investment valuation standards, one based on conventional cash-flow analysis, the other based on green-accounting standards is detailed in the paper. The analysis is performed as a part of the European Commission Framework Program Project AFTUR, undertaken by the consortium consisting of respectful European Research Establishments in renewable energy area as well as major European biofuel-powered turbine producers. The results show that the wider adoption of green accounting standards would induce the unprecedented growth of the renewable energy sector, because it would make the investment into renewable energy attractive for investors. (author)

  1. Green Roof Potential in Arab Cities

    OpenAIRE

    Attia, Shady

    2014-01-01

    Urban green roofs have long been promoted as an easy and effective strategy for beautifying the built environment and increasing investment opportunity. The building roof is very important because it has a direct impact on thermal comfort and energy conservation in and around buildings. Urban green roofs can help to address the lack of green space in many urban areas. Urban green roofs provides the city with open spaces that helps reduce urban heat island effect and provides the human populat...

  2. Challenges of Green Logistics in Southeast Europe

    OpenAIRE

    Beškovnik, Bojan; Jakomin, Livio

    2010-01-01

    This paper describes the trends towards green logistics in global aspect and challenges of adopting green logistics in the region of Southeast Europe. Modern logistics with supply chain management is experiencing a period of important evolution. From reversible logistics, we came to green logistics, which is a wider concept of environmentally friendly thinking. Reverse logistics includes processes of movements and transportation of waste from users to recycling plants; meanwhile, green logist...

  3. A novel genotoxin-specific qPCR array based on the metabolically competent human HepaRG™ cell line as a rapid and reliable tool for improved in vitro hazard assessment.

    Science.gov (United States)

    Ates, Gamze; Mertens, Birgit; Heymans, Anja; Verschaeve, Luc; Milushev, Dimiter; Vanparys, Philippe; Roosens, Nancy H C; De Keersmaecker, Sigrid C J; Rogiers, Vera; Doktorova, Tatyana Y

    2018-04-01

    Although the value of the regulatory accepted batteries for in vitro genotoxicity testing is recognized, they result in a high number of false positives. This has a major impact on society and industries developing novel compounds for pharmaceutical, chemical, and consumer products, as afflicted compounds have to be (prematurely) abandoned or further tested on animals. Using the metabolically competent human HepaRG ™ cell line and toxicogenomics approaches, we have developed an upgraded, innovative, and proprietary gene classifier. This gene classifier is based on transcriptomic changes induced by 12 genotoxic and 12 non-genotoxic reference compounds tested at sub-cytotoxic concentrations, i.e., IC10 concentrations as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The resulting gene classifier was translated into an easy-to-handle qPCR array that, as shown by pathway analysis, covers several different cellular processes related to genotoxicity. To further assess the predictivity of the tool, a set of 5 known positive and 5 known negative test compounds for genotoxicity was evaluated. In addition, 2 compounds with debatable genotoxicity data were tested to explore how the qPCR array would classify these. With an accuracy of 100%, when equivocal results were considered positive, the results showed that combining HepaRG ™ cells with a genotoxin-specific qPCR array can improve (geno)toxicological hazard assessment. In addition, the developed qPCR array was able to provide additional information on compounds for which so far debatable genotoxicity data are available. The results indicate that the new in vitro tool can improve human safety assessment of chemicals in general by basing predictions on mechanistic toxicogenomics information.

  4. 77 FR 24494 - Office of Federal High-Performance Green Buildings; Green Building Advisory Committee...

    Science.gov (United States)

    2012-04-24

    ... Federal High-Performance Green Buildings; Green Building Advisory Committee; Notification of Upcoming... agenda for the May 9, 2012, meeting of the Green Building Advisory Committee Meeting (the Committee). The... Sandler, Designated Federal Officer, Office of Federal High-Performance Green Buildings, Office of...

  5. 77 FR 66616 - Office of Federal High-Performance Green Buildings; Green Building Advisory Committee...

    Science.gov (United States)

    2012-11-06

    ... Federal High-Performance Green Buildings; Green Building Advisory Committee; Notification of Upcoming... and agenda for the November 27, 2012, meeting of the Green Building Advisory Committee Meeting (the... Green Buildings, Office of Government-wide Policy, General Services Administration, 1275 First Street NE...

  6. 78 FR 21368 - Office of Federal High-Performance Green Buildings; Green Building Advisory Committee...

    Science.gov (United States)

    2013-04-10

    ... Federal High-Performance Green Buildings; Green Building Advisory Committee; Notification of Upcoming... and agenda for the May 1, 2013, meeting of the Green Building Advisory Committee Meeting (the... Green Buildings, Office of Government-wide Policy, General Services Administration, 1275 First Street NE...

  7. Trends in Utility Green Pricing Programs (2005)

    Energy Technology Data Exchange (ETDEWEB)

    Bird, L.; Brown, E.

    2006-10-01

    This report presents year-end 2005 data on utility green pricing programs, and examines trends in consumer response and program implementation over time. The data in this report, which were obtained via a questionnaire distributed to utility green pricing program managers, can be used by utilities to benchmark the success of their green power programs.

  8. The Green Economy in the Global South

    DEFF Research Database (Denmark)

    Brockington, Dan; Ponte, Stefano

    2015-01-01

    As multiple visions for a Green Economy seek to become real, so are green economic initiatives in the global South multiplying. These can offer integration into wealth-generating markets – as well as displacement, alienation, conflict and opportunities for ‘green washing’. The articles included i...

  9. Green energy strategies for sustainable development

    International Nuclear Information System (INIS)

    Midilli, Adnan; Dincer, Ibrahim; Ay, Murat

    2006-01-01

    In this study we propose some green energy strategies for sustainable development. In this regard, seven green energy strategies are taken into consideration to determine the sectoral, technological, and application impact ratios. Based on these ratios, we derive a new parameter as the green energy impact ratio. In addition, the green energy-based sustainability ratio is obtained by depending upon the green energy impact ratio, and the green energy utilization ratio that is calculated using actual energy data taken from literature. In order to verify these parameters, three cases are considered. Consequently, it can be considered that the sectoral impact ratio is more important and should be kept constant as much as possible in a green energy policy implementation. Moreover, the green energy-based sustainability ratio increases with an increase of technological, sectoral, and application impact ratios. This means that all negative effects on the industrial, technological, sectoral and social developments partially and/or completely decrease throughout the transition and utilization to and of green energy and technologies when possible sustainable energy strategies are preferred and applied. Thus, the sustainable energy strategies can make an important contribution to the economies of the countries where green energy (e.g., wind, solar, tidal, biomass) is abundantly produced. Therefore, the investment in green energy supply and progress should be encouraged by governments and other authorities for a green energy replacement of fossil fuels for more environmentally benign and sustainable future

  10. Discussion on interior greening decoration of residence

    Science.gov (United States)

    Tong, ZhiNeng

    2018-02-01

    Green plants bring endless natural energy into the room. This paper introduces the functions, principles and functions of indoor greening decoration from different angles. Its conclusion is that it has become an important measure to improve the living environment of the people, and through the interior greening decoration to create a harmonious living space of architecture, human and nature.

  11. Green Roofs for Stormwater Runoff Control

    Science.gov (United States)

    This project evaluated green roofs as a stormwater management tool. Specifically, runoff quantity and quality from green and flat asphalt roofs were compared. Evapotranspiration from planted green roofs and evaporation from unplanted media roofs were also compared. The influence...

  12. Green power programs in Canada : 2003 : overview of Government green power policies, utility green power implementation initiatives, green power and certificate marketing programs, and their benefits

    International Nuclear Information System (INIS)

    Whitmore, J.; Bramley, M.; Holmes, R.

    2004-09-01

    Green power is defined as electricity produced from renewable sources, and whose production has low adverse impacts on the environment, human health and communities. Green power has near-zero greenhouse gas (GHG) emissions and includes sources such as wind, hydro, and solar power. It offers several environmental benefits, as well as the enhancement of energy security, regional development, economic diversification and the creation of skilled jobs. There are four categories of programs related to green power development in Canada: government green power policies, utility green power development programs, green power marketing initiatives, and green power certificate marketing initiatives. Most of the activities in Canada associated with these four categories in 2003 were discussed in this report. However, difficulties with quantification prevented the inclusion of some green power activities such as (1) the generation of green power not certified or identified by the generator as green power, (2) industry or residential self-generation, (3) net metering, and (4) small government programs. Green power generation facilities in 2003 totaled 775 MW of capacity compared to 539 MW in 2002. Hydro capacity represented 41 per cent, followed by wind capacity at 40 per cent and wood waste at 17 per cent. Most of the green power generation facilities in 2003 were located in Alberta, followed by British Columbia, Ontario and Quebec. 230 refs., 8 tabs., 1 fig

  13. Green chemistry education in the Middle East

    Science.gov (United States)

    Kolopajlo, Larry

    2017-06-01

    The Middle East once dominated the age of alchemy, and today it is experiencing a resurgence by transforming the age of petroleum chemicals into a greener science through Estidama. This green conversion is taking place through green chemical research and education. This report examines and reviews the understudied subject of green chemical education in the Middle East through the lens of context and history.

  14. CORRELATION BETWEEN CAFFEINE CONTENTS OF GREEN ...

    African Journals Online (AJOL)

    KEY WORDS: Green coffee beans, Caffeine, Correlation between caffeine content and altitude of coffee plant,. UV-Vis .... The extraction of caffeine from green coffee bean samples in to water was carried out by the reported method ..... caffeine in proposed green tea standard reference materials by liquid chromatography.

  15. Vitamin G : Green environments - Healthy environments

    NARCIS (Netherlands)

    Maas, J.

    2009-01-01

    The shortest summary of the thesis is in its title “Vitamin G”, where the G stands for the green space around us and Vitamin stands for the possible positive relationship between green space and people’s health. In the first part of this thesis we investigate whether green space in people’s living

  16. Closed forms for conformally flat Green's functions

    International Nuclear Information System (INIS)

    Brown, M.R.; Grove, P.G.; Ottewill, A.C.

    1981-01-01

    A closed form is obtained for the massless scalar Green's function on Rindler space. This is related by conformal transformation to the Green's function for a massless, conformally coupled scalar field on the open Einstein universe. A closed form is also obtained for the corresponding Green's function on the Einstein static universe. (author)

  17. Validation of chimerism in pediatric recipients of allogeneic hematopoietic stem cell transplantation (HSCT) a comparison between two methods: real-time PCR (qPCR) vs. variable number tandem repeats PCR (VNTR PCR).

    Science.gov (United States)

    Kletzel, Morris; Huang, Wei; Olszewski, Marie; Khan, Sana

    2013-01-01

    Post-hematopoietic stem cell transplantation (HSCT) chimerism monitoring is important to assess relapse and therapeutic intervention. The purpose of our study is to compare two methods variable number tandem repeats (VNTR) vs. quantitative real- time polymerase chain reaction (qPCR) in terms of determining chimerism. 127 (peripheral blood n=112, bone marrow n=15) samples were simultaneously tested by VNTR using APO-B, D1S80, D1S111, D17S30, gene loci SRY and ZP3 and qPCR using 34 assays (CA001-CA034) that are designed to a bi-allelic insertion/deletion (indel) polymorphism in the human genome. Samples were separated in three subsets: total WBC, T-cell and Myeloid cells. Extraction of DNA was performed then quantified. We analyzed column statistics, paired t-test and regression analysis for both methods. There was complete correlation between the two methods. The simplicity and rapidity of the test results from the qPCR method is more efficient and accurate to assess chimerism.

  18. RPL13A and EEF1A1 Are Suitable Reference Genes for qPCR during Adipocyte Differentiation of Vascular Stromal Cells from Patients with Different BMI and HOMA-IR.

    Science.gov (United States)

    Gentile, Adriana-Mariel; Lhamyani, Said; Coín-Aragüez, Leticia; Oliva-Olivera, Wilfredo; Zayed, Hatem; Vega-Rioja, Antonio; Monteseirin, Javier; Romero-Zerbo, Silvana-Yanina; Tinahones, Francisco-José; Bermúdez-Silva, Francisco-Javier; El Bekay, Rajaa

    2016-01-01

    Real-time or quantitative PCR (qPCR) is a useful technique that requires reliable reference genes for data normalization in gene expression analysis. Adipogenesis is among the biological processes suitable for this technique. The selection of adequate reference genes is essential for qPCR gene expression analysis of human Vascular Stromal Cells (hVSCs) during their differentiation into adipocytes. To the best of our knowledge, there are no studies validating reference genes for the analyses of visceral and subcutaneous adipose tissue hVSCs from subjects with different Body Mass Index (BMI) and Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) index. The present study was undertaken to analyze this question. We first analyzed the stability of expression of five potential reference genes: CYC, GAPDH, RPL13A, EEF1A1, and 18S ribosomal RNA, during in vitro adipogenic differentiation, in samples from these types of patients. The expression of RPL13A and EEF1A1 was not affected by differentiation, thus being these genes the most stable candidates, while CYC, GAPDH, and 18S were not suitable for this sort of analysis. This work highlights that RPL13A and EEF1A1 are good candidates as reference genes for qPCR analysis of hVSCs differentiation into adipocytes from subjects with different BMI and HOMA-IR.

  19. Evaluation of the efficiency of nested q-PCR in the detection of Mycobacterium tuberculosis complex directly from tuberculosis-suspected lesions in post-mortem macroscopic inspections of bovine carcasses slaughtered in the state of Mato Grosso, Brazil.

    Science.gov (United States)

    Carvalho, Ricardo César Tavares; Furlanetto, Leone Vinícius; Maruyama, Fernanda Harumy; Araújo, Cristina Pires de; Barros, Sílvia Letícia Bomfim; Ramos, Carlos Alberto do Nascimento; Dutra, Valéria; Araújo, Flábio Ribeiro de; Paschoalin, Vânia Margaret Flosi; Nakazato, Luciano; Figueiredo, Eduardo Eustáquio de Souza

    2015-08-01

    Bovine tuberculosis (BTB) is a zoonotic disease caused by Mycobacterium bovis, a member of the Mycobacterium tuberculosis complex (MTC). The quick and specific detection of this species is of extreme importance, since BTB may cause economic impacts, in addition to presenting imminent risks to human health. In the present study a nested real-time PCR test (nested q-PCR) was used in post-mortem evaluations to assess cattle carcasses with BTB-suspected lesions. A total of 41,193 cattle slaughtered in slaughterhouses located in the state of Mato Grosso, were examined. Of the examined animals, 198 (0.48%) showed BTB-suspected lesions. M. bovis was isolated in 1.5% (3/198) of the samples. Multiplex-PCR detected MTC in 7% (14/198) of the samples. The nested q-PCR test detected MTC in 28% (56/198) of the BTB-suspected lesions, demonstrating higher efficiency when compared to the multiplex-PCR and conventional microbiology. Nested q-PCR can therefore be used as a complementary test in the national program for control and eradication of bovine tuberculosis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Green-E general program and public information support program report, August 1, 1999 - September 30, 2000

    Energy Technology Data Exchange (ETDEWEB)

    Brown, Kirk

    2000-09-30

    Green-E Program support from the Dept. of Energy augmented the costs of implementing the objectives of the Green-E Renewable Electricity Project; general program implementation; regional adaptation; developing strategic partnerships; and public information/education/outreach.