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 ...

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

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.

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

Science.gov (United States)

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.

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…

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.

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.

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.

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.

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

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.

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.

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.

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.

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

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.

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...

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.

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

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%.

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

Science.gov (United States)

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.

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.

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....

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.

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.

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.

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.

Development of Quantitative Real-Time PCR Assays for Detection and Quantification of Surrogate Biological Warfare Agents in Building Debris and Leachate▿

Science.gov (United States)

Saikaly, Pascal E.; Barlaz, Morton A.; de los Reyes, Francis L.

2007-01-01

Evaluation of the fate and transport of biological warfare (BW) agents in landfills requires the development of specific and sensitive detection assays. The objective of the current study was to develop and validate SYBR green quantitative real-time PCR (Q-PCR) assays for the specific detection and quantification of surrogate BW agents in synthetic building debris (SBD) and leachate. Bacillus atrophaeus (vegetative cells and spores) and Serratia marcescens were used as surrogates for Bacillus anthracis (anthrax) and Yersinia pestis (plague), respectively. The targets for SYBR green Q-PCR assays were the 16S-23S rRNA intergenic transcribed spacer (ITS) region and recA gene for B. atrophaeus and the gyrB, wzm, and recA genes for S. marcescens. All assays showed high specificity when tested against 5 ng of closely related Bacillus and Serratia nontarget DNA from 21 organisms. Several spore lysis methods that include a combination of one or more of freeze-thaw cycles, chemical lysis, hot detergent treatment, bead beat homogenization, and sonication were evaluated. All methods tested showed similar threshold cycle values. The limit of detection of the developed Q-PCR assays was determined using DNA extracted from a pure bacterial culture and DNA extracted from sterile water, leachate, and SBD samples spiked with increasing quantities of surrogates. The limit of detection for B. atrophaeus genomic DNA using the ITS and B. atrophaeus recA Q-PCR assays was 7.5 fg per PCR. The limits of detection of S. marcescens genomic DNA using the gyrB, wzm, and S. marcescens recA Q-PCR assays were 7.5 fg, 75 fg, and 7.5 fg per PCR, respectively. Quantification of B. atrophaeus vegetative cells and spores was linear (R2 > 0.98) over a 7-log-unit dynamic range down to 101 B. atrophaeus cells or spores. Quantification of S. marcescens (R2 > 0.98) was linear over a 6-log-unit dynamic range down to 102 S. marcescens cells. The developed Q-PCR assays are highly specific and sensitive and can

Quantitative microbial community analysis of three different sulfidic mine tailing dumps generating acid mine drainage.

Science.gov (United States)

Kock, Dagmar; Schippers, Axel

2008-08-01

The microbial communities of three different sulfidic and acidic mine waste tailing dumps located in Botswana, Germany, and Sweden were quantitatively analyzed using quantitative real-time PCR (Q-PCR), fluorescence in situ hybridization (FISH), catalyzed reporter deposition-FISH (CARD-FISH), Sybr green II direct counting, and the most probable number (MPN) cultivation technique. Depth profiles of cell numbers showed that the compositions of the microbial communities are greatly different at the three sites and also strongly varied between zones of oxidized and unoxidized tailings. Maximum cell numbers of up to 10(9) cells g(-1) dry weight were determined in the pyrite or pyrrhotite oxidation zones, whereas cell numbers in unoxidized tailings were significantly lower. Bacteria dominated over Archaea and Eukarya at all tailing sites. The acidophilic Fe(II)- and/or sulfur-oxidizing Acidithiobacillus spp. dominated over the acidophilic Fe(II)-oxidizing Leptospirillum spp. among the Bacteria at two sites. The two genera were equally abundant at the third site. The acidophilic Fe(II)- and sulfur-oxidizing Sulfobacillus spp. were generally less abundant. The acidophilic Fe(III)-reducing Acidiphilium spp. could be found at only one site. The neutrophilic Fe(III)-reducing Geobacteraceae as well as the dsrA gene of sulfate reducers were quantifiable at all three sites. FISH analysis provided reliable data only for tailing zones with high microbial activity, whereas CARD-FISH, Q-PCR, Sybr green II staining, and MPN were suitable methods for a quantitative microbial community analysis of tailings in general.

Properties of targeted preamplification in DNA and cDNA quantification.

Science.gov (United States)

Andersson, Daniel; Akrap, Nina; Svec, David; Godfrey, Tony E; Kubista, Mikael; Landberg, Göran; Ståhlberg, Anders

2015-01-01

Quantification of small molecule numbers often requires preamplification to generate enough copies for accurate downstream enumerations. Here, we studied experimental parameters in targeted preamplification and their effects on downstream quantitative real-time PCR (qPCR). To evaluate different strategies, we monitored the preamplification reaction in real-time using SYBR Green detection chemistry followed by melting curve analysis. Furthermore, individual targets were evaluated by qPCR. The preamplification reaction performed best when a large number of primer pairs was included in the primer pool. In addition, preamplification efficiency, reproducibility and specificity were found to depend on the number of template molecules present, primer concentration, annealing time and annealing temperature. The amount of nonspecific PCR products could also be reduced about 1000-fold using bovine serum albumin, glycerol and formamide in the preamplification. On the basis of our findings, we provide recommendations how to perform robust and highly accurate targeted preamplification in combination with qPCR or next-generation sequencing.

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.

Identification of subtelomeric genomic imbalances and breakpoint mapping with quantitative PCR in 296 individuals with congenital defects and/or mental retardation

Directory of Open Access Journals (Sweden)

Brockmann Knut

2009-03-01

Full Text Available Abstract Background Submicroscopic imbalances in the subtelomeric regions of the chromosomes are considered to play an important role in the aetiology of mental retardation (MR. The aim of the study was to evaluate a quantitative PCR (qPCR protocol established by Boehm et al. (2004 in the clinical routine of subtelomeric testing. Results 296 patients with MR and a normal karyotype (500–550 bands were screened for subtelomeric imbalances by using qPCR combined with SYBR green detection. In total, 17 patients (5.8% with 20 subtelomeric imbalances were identified. Six of the aberrations (2% were classified as causative for the symptoms, because they occurred either de novo in the patients (5 cases or the aberration were be detected in the patient and an equally affected parent (1 case. The extent of the deletions ranged from 1.8 to approximately 10 Mb, duplications were 1.8 to approximately 5 Mb in size. In 6 patients, the copy number variations (CNVs were rated as benign polymorphisms, and the clinical relevance of these CNVs remains unclear in 5 patients (1.7%. Therefore, the overall frequency of clinically relevant imbalances ranges between 2% and 3.7% in our cohort. Conclusion This study illustrates that the qPCR/SYBR green technique represents a rapid and versatile method for the detection of subtelomeric imbalances and the option to map the breakpoint. Thus, this technique is highly suitable for genotype/phenotype studies in patients with MR/developmental delay and/or congenital defects.

Upconversion luminescence resonance energy transfer-based aptasensor for the sensitive detection of oxytetracycline.

Science.gov (United States)

Zhang, Hui; Fang, Congcong; Wu, Shijia; Duan, Nuo; Wang, Zhouping

2015-11-15

In this work, a biosensor based on luminescence resonance energy transfer (LRET) from NaYF4:Yb,Tm upconversion nanoparticles (UCNPs) to SYBR Green I has been developed. The aptamers are covalently linked to UCNPs and hybridized with their complementary strands. The subsequent addition of SYBR Green allows SYBR Green I to insert into the formed double-stranded DNA (dsDNA) duplex and brings the energy donor and acceptor into close proximity, leading to the fluorescence of UCNPs transferred to SYBR Green I. When excited at 980 nm, the UCNPs emit luminescence at 477 nm, and this energy is transferred to SYBR Green I, which emits luminescence at 530 nm. In the presence of oxytetracycline (OTC), the aptamers prefer to bind to its corresponding analyte and dehybridize with the complementary DNA. This dehybridization leads to the liberation of SYBR Green I, which distances SYBR Green I from the UCNPs and recovers the UCNPs' luminescence. Under optimal conditions, a linear calibration is obtained between the ratio of I530 to I477 nm (I530/I477) and the OTC concentration, which ranges from 0.1 to 10 ng/ml with a limit of detection (LOD) of 0.054 ng/ml. Copyright © 2015 Elsevier Inc. All rights reserved.

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...

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.

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.

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.

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.

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.

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

Fluorometric determination of nucleic acids based on the use of polydopamine nanotubes and target-induced strand displacement amplification.

Science.gov (United States)

Ge, Jia; Bai, Dong-Mei; -Geng, Xin; Hu, Ya-Lei; Cai, Qi-Yong; Xing, Ke; Zhang, Lin; Li, Zhao-Hui

2018-01-10

The authors describe a fluorometric method for the quantitation of nucleic acids by combining (a) cycled strand displacement amplification, (b) the unique features of the DNA probe SYBR Green, and (c) polydopamine nanotubes. SYBR Green undergoes strong fluorescence enhancement upon intercalation into double-stranded DNA (dsDNA). The polydopamine nanotubes selectively adsorb single-stranded DNA (ssDNA) and molecular beacons. In the absence of target DNA, the molecular beacon, primer and SYBR Green are adsorbed on the surface of polydopamine nanotubes. This results in quenching of the fluorescence of SYBR Green, typically measured at excitation/emission wavelengths of 488/518 nm. Upon addition of analyte (target DNA) and polymerase, the stem of the molecular beacon is opened so that it can bind to the primer. This triggers target strand displacement polymerization, during which dsDNA is synthesized. The hybridized target is then displaced due to the strand displacement activity of the polymerase. The displaced target hybridizes with another molecular beacon. This triggers the next round of polymerization. Consequently, a large amount of dsDNA is formed which is detected by addition of SYBR Green. Thus, sensitive and selective fluorometric detection is realized. The fluorescent sensing strategy shows very good analytical performances towards DNA detection, such as a wide linear range from 0.05 to 25 nM with a low limit of detection of 20 pM. Graphical abstract Schematic of a fluorometric strategy for highly sensitive and selective determination of nucleic acids by combining strand displacement amplification and the unique features of SYBR Green I (SG) and polydopamine nanotubes.

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.

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.

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.

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.

A comprehensive collection of experimentally validated primers for Polymerase Chain Reaction quantitation of murine transcript abundance

Directory of Open Access Journals (Sweden)

Wang Xiaowei

2008-12-01

Full Text Available Abstract Background Quantitative polymerase chain reaction (QPCR is a widely applied analytical method for the accurate determination of transcript abundance. Primers for QPCR have been designed on a genomic scale but non-specific amplification of non-target genes has frequently been a problem. Although several online databases have been created for the storage and retrieval of experimentally validated primers, only a few thousand primer pairs are currently present in existing databases and the primers are not designed for use under a common PCR thermal profile. Results We previously reported the implementation of an algorithm to predict PCR primers for most known human and mouse genes. We now report the use of that resource to identify 17483 pairs of primers that have been experimentally verified to amplify unique sequences corresponding to distinct murine transcripts. The primer pairs have been validated by gel electrophoresis, DNA sequence analysis and thermal denaturation profile. In addition to the validation studies, we have determined the uniformity of amplification using the primers and the technical reproducibility of the QPCR reaction using the popular and inexpensive SYBR Green I detection method. Conclusion We have identified an experimentally validated collection of murine primer pairs for PCR and QPCR which can be used under a common PCR thermal profile, allowing the evaluation of transcript abundance of a large number of genes in parallel. This feature is increasingly attractive for confirming and/or making more precise data trends observed from experiments performed with DNA microarrays.

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

Inhibition of miR-1247 on cell proliferation and invasion in bladder ...

Indian Academy of Sciences (India)

Yudi Zhu

2018-04-21

Apr 21, 2018 ... the best of our knowledge, the relationship between miR-. 1247 and bladder cancer ... justify the anti-tumour function of this gene. Although the ... miR-1247 was car- ried out by using Power SYBR qPCR and miRNA qRT-PCR.

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.

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.

Detection and persistence of fecal Bacteroidales as water quality indicators in unchlorinated drinking water

DEFF Research Database (Denmark)

Saunders, Aaron Marc; Kristiansen, Anja; Lund, Marie Braad

2009-01-01

doi:10.1016/j.syapm.2008.11.004 The results of this study support the use of fecal Bacteroidales qPCR as a rapid method to complement traditional, culture dependent, water quality indicators in systems where drinking water is supplied without chlorination or other forms of disinfection. A SYBR...... green based, quantitative PCR assay was developed to determine the concentration of fecal Bacteroidales 16S rRNA gene copies. The persistence of a Bacteroides vulgatus pure culture and fecal Bacteroidales from a wastewater inoculum was determined in unchlorinated drinking water at10°C. B. vulgatus 16S r......RNA gene copies persisted throughout the experimental period (200 days) in sterile drinking water but decayed faster in natural drinking water, indicating that the natural microbiota accelerated decay. In a simulated fecal contamination of unchlorinated drinking water, the decay of fecal Bacteroidales 16S...

Comparison of Suitability of the Most Common Ancient DNA Quantification Methods.

Science.gov (United States)

Brzobohatá, Kristýna; Drozdová, Eva; Smutný, Jiří; Zeman, Tomáš; Beňuš, Radoslav

2017-04-01

Ancient DNA (aDNA) extracted from historical bones is damaged and fragmented into short segments, present in low quantity, and usually copurified with microbial DNA. A wide range of DNA quantification methods are available. The aim of this study was to compare the five most common DNA quantification methods for aDNA. Quantification methods were tested on DNA extracted from skeletal material originating from an early medieval burial site. The tested methods included ultraviolet (UV) absorbance, real-time quantitative polymerase chain reaction (qPCR) based on SYBR ® green detection, real-time qPCR based on a forensic kit, quantification via fluorescent dyes bonded to DNA, and fragmentary analysis. Differences between groups were tested using a paired t-test. Methods that measure total DNA present in the sample (NanoDrop ™ UV spectrophotometer and Qubit ® fluorometer) showed the highest concentrations. Methods based on real-time qPCR underestimated the quantity of aDNA. The most accurate method of aDNA quantification was fragmentary analysis, which also allows DNA quantification of the desired length and is not affected by PCR inhibitors. Methods based on the quantification of the total amount of DNA in samples are unsuitable for ancient samples as they overestimate the amount of DNA presumably due to the presence of microbial DNA. Real-time qPCR methods give undervalued results due to DNA damage and the presence of PCR inhibitors. DNA quantification methods based on fragment analysis show not only the quantity of DNA but also fragment length.

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

Comparison of multiple DNA dyes for real-time PCR: effects of dye concentration and sequence composition on DNA amplification and melting temperature

DEFF Research Database (Denmark)

Guðnason, Haukur; Dufva, Hans Martin; Bang, Dang Duong

2007-01-01

investigate 15 different intercalating DNA dyes for their inhibitory effects on PCR, effects on DNA melting temperature and possible preferential binding to GC-rich sequences. Our results demonstrated that in contrast to the results of SYBR Green I, two intercalating dyes SYTO-13 and SYTO-82 do not inhibit......The importance of real-time polymerase chain reaction (PCR) has increased steadily in clinical applications over the last decade. Many applications utilize SYBR Green I dye to follow the accumulation of amplicons in real time. SYBR Green I has, however, a number of limitations that include...... the inhibition of PCR, preferential binding to GC-rich sequences and effects on melting curve analysis. Although a few alternative dyes without some of these limitations have been recently proposed, no large-scale investigation into the properties of intercalating dyes has been performed. In this study, we...

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.

A Quantitative Polymerase Chain Reaction Assay for the Detection and Quantification of Epizootic Epitheliotropic Disease Virus (EEDV; Salmonid Herpesvirus 3).

Science.gov (United States)

Glenney, Gavin W; Barbash, Patricia A; Coll, John A

2016-03-01

Epizootic epitheliotropic disease virus (EEDV; salmonid herpesvirus [SalHV3]; family Alloherpesviridae) causes a systemic disease of juvenile and yearling Lake Trout Salvelinus namaycush. No cell lines are currently available for the culture and propagation of EEDV, so primary diagnosis is limited to PCR and electron microscopy. To better understand the pervasiveness of EEDV (carrier or latent state of infection) in domesticated and wild Lake Trout populations, we developed a sensitive TaqMan quantitative PCR (qPCR) assay to detect the presence of the EEDV terminase gene in Lake Trout tissues. This assay was able to detect a linear standard curve over nine logs of plasmid dilution and was sensitive enough to detect single-digit copies of EEDV. The efficiency of the PCR assay was 99.4 ± 0.06% (mean ± SD), with a 95% confidence limit of 0.0296 (R(2) = 0.994). Methods were successfully applied to collect preliminary data from a number of species and water bodies in the states of Pennsylvania, New York, and Vermont, indicating that EEDV is more common in wild fish than previously known. In addition, through the development of this qPCR assay, we detected EEDV in a new salmonid species, the Cisco Coregonus artedi. The qPCR assay was unexpectedly able to detect two additional herpesviruses, the Atlantic Salmon papillomatosis virus (ASPV; SalHV4) and the Namaycush herpesvirus (NamHV; SalHV5), which both share high sequence identity with the EEDV terminase gene. With these unexpected findings, we subsequently designed three primer sets to confirm initial TaqMan qPCR assay positives and to differentiate among EEDV, ASPV, and NamHV by detecting the glycoprotein genes via SYBR Green qPCR. Received April 20, 2015; accepted November 10, 2015.

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.

Quantitative monitoring of microbial species during bioleaching of a copper concentrate

Directory of Open Access Journals (Sweden)

Sabrina Hedrich

2016-12-01

Full Text Available Monitoring of the microbial community in bioleaching processes is essential in order to control process parameters and enhance the leaching efficiency. Suitable methods are, however, limited as they are usually not adapted to bioleaching samples and often no taxon-specific assays are available in the literature for these types of consortia. Therefore, our study focused on the development of novel quantitative real-time PCR (qPCR assays for the quantification of Acidithiobacillus caldus, Leptospirillum ferriphilum, Sulfobacillus thermosulfidooxidans and Sulfobacillus benefaciens and comparison of the results with data from other common molecular monitoring methods in order to evaluate their accuracy and specificity. Stirred tank bioreactors for the leaching of copper concentrate, housing a consortium of acidophilic, moderately-thermophilic bacteria, relevant in several bioleaching operations, served as a model system. The microbial community analysis via qPCR allowed a precise monitoring of the evolution of total biomass as well as abundance of specific species. Data achieved by the standard fingerprinting methods, terminal restriction fragment length polymorphism (T-RFLP and capillary electrophoresis single strand conformation polymorphism (CE-SSCP on the same samples followed the same trend as qPCR data. The main added value of qPCR was, however, to provide quantitative data for each species whereas only relative abundance could be deduced from T-RFLP and CE-SSCP profiles. Additional value was obtained by applying two further quantitative methods which do not require nucleic acid extraction, total cell counting after SYBR Green staining and metal sulfide oxidation activity measurements via microcalorimetry. Overall, these complementary methods allow for an efficient quantitative microbial community monitoring in various bioleaching operations.

Quantitative Monitoring of Microbial Species during Bioleaching of a Copper Concentrate.

Science.gov (United States)

Hedrich, Sabrina; Guézennec, Anne-Gwenaëlle; Charron, Mickaël; Schippers, Axel; Joulian, Catherine

2016-01-01

Monitoring of the microbial community in bioleaching processes is essential in order to control process parameters and enhance the leaching efficiency. Suitable methods are, however, limited as they are usually not adapted to bioleaching samples and often no taxon-specific assays are available in the literature for these types of consortia. Therefore, our study focused on the development of novel quantitative real-time PCR (qPCR) assays for the quantification of Acidithiobacillus caldus, Leptospirillum ferriphilum, Sulfobacillus thermosulfidooxidans , and Sulfobacillus benefaciens and comparison of the results with data from other common molecular monitoring methods in order to evaluate their accuracy and specificity. Stirred tank bioreactors for the leaching of copper concentrate, housing a consortium of acidophilic, moderately thermophilic bacteria, relevant in several bioleaching operations, served as a model system. The microbial community analysis via qPCR allowed a precise monitoring of the evolution of total biomass as well as abundance of specific species. Data achieved by the standard fingerprinting methods, terminal restriction fragment length polymorphism (T-RFLP) and capillary electrophoresis single strand conformation polymorphism (CE-SSCP) on the same samples followed the same trend as qPCR data. The main added value of qPCR was, however, to provide quantitative data for each species whereas only relative abundance could be deduced from T-RFLP and CE-SSCP profiles. Additional value was obtained by applying two further quantitative methods which do not require nucleic acid extraction, total cell counting after SYBR Green staining and metal sulfide oxidation activity measurements via microcalorimetry. Overall, these complementary methods allow for an efficient quantitative microbial community monitoring in various bioleaching operations.

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

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.

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.

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

Ultrasensitive electrochemical biosensor for detection of DNA from Bacillus subtilis by coupling target-induced strand displacement and nicking endonuclease signal amplification.

Science.gov (United States)

Hu, Yuhua; Xu, Xueqin; Liu, Qionghua; Wang, Ling; Lin, Zhenyu; Chen, Guonan

2014-09-02

A simple, ultrasensitive, and specific electrochemical biosensor was designed to determine the given DNA sequence of Bacillus subtilis by coupling target-induced strand displacement and nicking endonuclease signal amplification. The target DNA (TD, the DNA sequence from the hypervarient region of 16S rDNA of Bacillus subtilis) could be detected by the differential pulse voltammetry (DPV) in a range from 0.1 fM to 20 fM with the detection limit down to 0.08 fM at the 3s(blank) level. This electrochemical biosensor exhibits high distinction ability to single-base mismatch, double-bases mismatch, and noncomplementary DNA sequence, which may be expected to detect single-base mismatch and single nucleotide polymorphisms (SNPs). Moreover, the applicability of the designed biosensor for detecting the given DNA sequence from Bacillus subtilis was investigated. The result obtained by electrochemical method is approximately consistent with that by a real-time quantitative polymerase chain reaction detecting system (QPCR) with SYBR Green.

Diagnosis of bacteremia in pediatric oncologic patients by in-house real-time PCR.

Science.gov (United States)

Quiles, Milene Gonçalves; Menezes, Liana Carballo; Bauab, Karen de Castro; Gumpl, Elke Kreuscher; Rocchetti, Talita Trevizani; Palomo, Flavia Silva; Carlesse, Fabianne; Pignatari, Antonio Carlos Campos

2015-07-23

Infections are the major cause of morbidity and mortality in children with cancer. Gaining a favorable prognosis for these patients depends on selecting the appropriate therapy, which in turn depends on rapid and accurate microbiological diagnosis. This study employed real-time PCR (qPCR) to identify the main pathogens causing bloodstream infection (BSI) in patients treated at the Pediatric Oncology Institute IOP-GRAACC-UNIFESP-Brazil. Antimicrobial resistance genes were also investigated using this methodology. A total of 248 samples from BACTEC® blood culture bottles and 99 whole-blood samples collected in tubes containing EDTA K2 Gel were isolated from 137 patients. All samples were screened by specific Gram probes for multiplex qPCR. Seventeen sequences were evaluated using gender-specific TaqMan probes and the resistance genes bla SHV, bla TEM, bla CTX, bla KPC, bla IMP, bla SPM, bla VIM, vanA, vanB and mecA were detected using the SYBR Green method. Positive qPCR results were obtained in 112 of the blood culture bottles (112/124), and 90 % agreement was observed between phenotypic and molecular microbial detection methods. For bacterial and fungal identification, the performance test showed: sensitivity 87 %; specificity 91 %; NPV 90 %; PPV 89 % and accuracy of 89 % when compared with the phenotypic method. The mecA gene was detected in 37 samples, extended-spectrum β-lactamases were detected in six samples and metallo-β-lactamase coding genes in four samples, with 60 % concordance between the two methods. The qPCR on whole blood detected eight samples possessing the mecA gene and one sample harboring the vanB gene. The bla KPC, bla VIM, bla IMP and bla SHV genes were not detected in this study. Real-time PCR is a useful tool in the early identification of pathogens and antimicrobial resistance genes from bloodstream infections of pediatric oncologic patients.

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

Pitfalls of DNA Quantification Using DNA-Binding Fluorescent Dyes and Suggested Solutions.

Science.gov (United States)

Nakayama, Yuki; Yamaguchi, Hiromi; Einaga, Naoki; Esumi, Mariko

2016-01-01

The Qubit fluorometer is a DNA quantification device based on the fluorescence intensity of fluorescent dye binding to double-stranded DNA (dsDNA). Qubit is generally considered useful for checking DNA quality before next-generation sequencing because it measures intact dsDNA. To examine the most accurate and suitable methods for quantifying DNA for quality assessment, we compared three quantification methods: NanoDrop, which measures UV absorbance; Qubit; and quantitative PCR (qPCR), which measures the abundance of a target gene. For the comparison, we used three types of DNA: 1) DNA extracted from fresh frozen liver tissues (Frozen-DNA); 2) DNA extracted from formalin-fixed, paraffin-embedded liver tissues comparable to those used for Frozen-DNA (FFPE-DNA); and 3) DNA extracted from the remaining fractions after RNA extraction with Trizol reagent (Trizol-DNA). These DNAs were serially diluted with distilled water and measured using three quantification methods. For Frozen-DNA, the Qubit values were not proportional to the dilution ratio, in contrast with the NanoDrop and qPCR values. This non-proportional decrease in Qubit values was dependent on a lower salt concentration, and over 1 mM NaCl in the DNA solution was required for the Qubit measurement. For FFPE-DNA, the Qubit values were proportional to the dilution ratio and were lower than the NanoDrop values. However, electrophoresis revealed that qPCR reflected the degree of DNA fragmentation more accurately than Qubit. Thus, qPCR is superior to Qubit for checking the quality of FFPE-DNA. For Trizol-DNA, the Qubit values were proportional to the dilution ratio and were consistently lower than the NanoDrop values, similar to FFPE-DNA. However, the qPCR values were higher than the NanoDrop values. Electrophoresis with SYBR Green I and single-stranded DNA (ssDNA) quantification demonstrated that Trizol-DNA consisted mostly of non-fragmented ssDNA. Therefore, Qubit is not always the most accurate method for

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.

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.

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...

Quantification of Listeria monocytogenes in minimally processed leafy vegetables using a combined method based on enrichment and 16S rRNA real-time PCR.

Science.gov (United States)

Aparecida de Oliveira, Maria; Abeid Ribeiro, Eliana Guimarães; Morato Bergamini, Alzira Maria; Pereira De Martinis, Elaine Cristina

2010-02-01

Modern lifestyle markedly changed eating habits worldwide, with an increasing demand for ready-to-eat foods, such as minimally processed fruits and leafy greens. Packaging and storage conditions of those products may favor the growth of psychrotrophic bacteria, including the pathogen Listeria monocytogenes. In this work, minimally processed leafy vegetables samples (n = 162) from retail market from Ribeirão Preto, São Paulo, Brazil, were tested for the presence or absence of Listeria spp. by the immunoassay Listeria Rapid Test, Oxoid. Two L. monocytogenes positive and six artificially contaminated samples of minimally processed leafy vegetables were evaluated by the Most Probable Number (MPN) with detection by classical culture method and also culture method combined with real-time PCR (RTi-PCR) for 16S rRNA genes of L. monocytogenes. Positive MPN enrichment tubes were analyzed by RTi-PCR with primers specific for L. monocytogenes using the commercial preparation ABSOLUTE QPCR SYBR Green Mix (ABgene, UK). Real-time PCR assay presented good exclusivity and inclusivity results and no statistical significant difference was found in comparison with the conventional culture method (p < 0.05). Moreover, RTi-PCR was fast and easy to perform, with MPN results obtained in ca. 48 h for RTi-PCR in comparison to 7 days for conventional method.

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.

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

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

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.

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.

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.

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.

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...

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.

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

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.

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

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.

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.

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...

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.

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…

Shining a light on LAMP assays--a comparison of LAMP visualization methods including the novel use of berberine.

Science.gov (United States)

Fischbach, Jens; Xander, Nina Carolin; Frohme, Marcus; Glökler, Jörn Felix

2015-04-01

The need for simple and effective assays for detecting nucleic acids by isothermal amplification reactions has led to a great variety of end point and real-time monitoring methods. Here we tested direct and indirect methods to visualize the amplification of potato spindle tuber viroid (PSTVd) by loop-mediated isothermal amplification (LAMP) and compared features important for one-pot in-field applications. We compared the performance of magnesium pyrophosphate, hydroxynaphthol blue (HNB), calcein, SYBR Green I, EvaGreen, and berberine. All assays could be used to distinguish between positive and negative samples in visible or UV light. Precipitation of magnesium-pyrophosphate resulted in a turbid reaction solution. The use of HNB resulted in a color change from violet to blue, whereas calcein induced a change from orange to yellow-green. We also investigated berberine as a nucleic acid-specific dye that emits a fluorescence signal under UV light after a positive LAMP reaction. It has a comparable sensitivity to SYBR Green I and EvaGreen. Based on our results, an optimal detection method can be chosen easily for isothermal real-time or end point screening applications.

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

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.

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...

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...

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

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.

Association between VDAC1 mRNA expression and intracellular ...

African Journals Online (AJOL)

ELO

2012-01-05

Jan 5, 2012 ... generate reactive oxygen species (ROS) and oxidative stress. Unchecked ... PCR Master Mix, SYBR Green-Real Master Mix, and Quantscript ... had reached 70%, they were exposed to Cr(VI) for different time periods (12, 24 ...

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...

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.

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...

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.

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.

MicroRNAs and cardiac sarcoplasmic reticulum calcium ATPase-2 in human myocardial infarction: expression and bioinformatic analysis.

Science.gov (United States)

Boštjančič, Emanuela; Zidar, Nina; Glavač, Damjan

2012-10-15

Cardiac sarco(endo)plasmic reticulum calcium ATPase-2 (SERCA2) plays one of the central roles in myocardial contractility. Both, SERCA2 mRNA and protein are reduced in myocardial infarction (MI), but the correlation has not been always observed. MicroRNAs (miRNAs) act by targeting 3'-UTR mRNA, causing translational repression in physiological and pathological conditions, including cardiovascular diseases. One of the aims of our study was to identify miRNAs that could influence SERCA2 expression in human MI. The protein SERCA2 was decreased and 43 miRNAs were deregulated in infarcted myocardium compared to corresponding remote myocardium, analyzed by western blot and microRNA microarrays, respectively. All the samples were stored as FFPE tissue and in RNAlater. miRNAs binding prediction to SERCA2 including four prediction algorithms (TargetScan, PicTar, miRanda and mirTarget2) identified 213 putative miRNAs. TAM and miRNApath annotation of deregulated miRNAs identified 18 functional and 21 diseased states related to heart diseases, and association of the half of the deregulated miRNAs to SERCA2. Free-energy of binding and flanking regions (RNA22, RNAfold) was calculated for 10 up-regulated miRNAs from microarray analysis (miR-122, miR-320a/b/c/d, miR-574-3p/-5p, miR-199a, miR-140, and miR-483), and nine miRNAs deregulated from microarray analysis were used for validation with qPCR (miR-21, miR-122, miR-126, miR-1, miR-133, miR-125a/b, and miR-98). Based on qPCR results, the comparison between FFPE and RNAlater stored tissue samples, between Sybr Green and TaqMan approaches, as well as between different reference genes were also performed. Combing all the results, we identified certain miRNAs as potential regulators of SERCA2; however, further functional studies are needed for verification. Using qPCR, we confirmed deregulation of nine miRNAs in human MI, and show that qPCR normalization strategy is important for the outcome of miRNA expression analysis in human MI.

Study of changes in bacterial and viral abundance in formaldehyde - Fixed water samples by epifluorescence microscopy

Digital Repository Service at National Institute of Oceanography (India)

Parvathi, A.; Radhakrishnan, S.; Sajila, M.P.; Jacob, B.

of bacteria and viruses in water samples from Cochin Backwater was determined by SYBR Green I staining and epifluorescence microscopy. The counts were determined for 45 days in samples fixed with 1–6% formaldehyde. The results suggest rapid decline in counts...

Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data

NARCIS (Netherlands)

Ramakers, Christian; Ruijter, Jan M.; Deprez, Ronald H. Lekanne; Moorman, Antoon F. M.

2003-01-01

Quantification of mRNAs using real-time polymerase chain reaction (PCR) by monitoring the product formation with the fluorescent dye SYBR Green I is being extensively used in neurosciences, developmental biology, and medical diagnostics. Most PCR data analysis procedures assume that the PCR

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.

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.

Molecular detection of Xanthomonas oryzae pv. oryzae, Xanthomonas oryzae pv. oryzicola, and Burkholderia glumae in infected rice seeds and leaves

Science.gov (United States)

Polymerase chain reaction (PCR) is particularly useful for plant pathogen detection. In the present study, multiplex PCR and SYBR green real-time PCR were developed to facilitate simultaneous detection of three important rice pathogens, Xanthomonas oryzae pv. oryzae, X. oryzae pv. oryzicola, and Bur...

Identification of chikungunya virus interacting proteins in mammalian ...

Indian Academy of Sciences (India)

2014-05-01

May 1, 2014 ... Chikungunya virus (CHIKV) is an alphavirus transmitted by. Aedes mosquitoes .... Knockdown of HSP70 (NM_005346, X70684) and STAT-2 ... Large scale endotoxin free plasmids were ... Biosystems, USA) using power SYBR Green I technology .... At 12 h p.i., pre-incubation with higher concentration of.

Effects of ovariectomization (OVX) and Synovex-Plus on the ...

African Journals Online (AJOL)

Clayton Bailey

g/kg; cobalt carbonate, 0.02 g/kg; copper sulphate, 1.57 g/kg; iron sulphate, 1.33 g/kg; ... and exsanguination, beginning at 07:00, at the University of Arizona Meats ... sample well on every plate and all samples were run with iTaq SYBR Green ...

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.

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

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.

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.

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.

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.

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.

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

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...

TCF7L2 expression in diabetic patients undergoing bariatric surgery.

Science.gov (United States)

Hindle, A Katharine; Brody, Fred; Tevar, Rahul; Kluk, Brian; Hill, Sarah; McCaffrey, Timothy; Fu, Sidney

2009-04-01

The cause of diabetes in morbidly obese patients is multifactorial, including genetic, social, and dietary components. Transcription factor 7-like 2 (TCF7L2) is a gene that is related to the development of diabetes. This pilot study examines TCF7L2 expression in liver samples obtained from morbidly obese patients undergoing bariatric surgery. TCF7L2 expression is compared between diabetic and nondiabetic patients. Liver samples were obtained from 20 morbidly obese patients undergoing bariatric surgery. Samples were flash frozen in liquid nitrogen. Total RNA was extracted from tissue samples using the TRIzol reagent (Invitrogen Inc, Carlsbad, CA). Using the iScript cDNA synthesis kit (Bio-Rad Laboratories, Hercules,CA), cDNA was synthesized. Quantitative polymerase chain reaction (qPCR) was done using SYBR Green qPCR Reagents (Stratagene, Cedar Creek TX) and the 7300 Real-Time PCR system (Applied Biosystems, Foster City CA). Preoperative demographic and gene expression data were correlated using univariate analysis and logistic regression models. Only associations with a p-value less than 0.05 were considered significant. For the entire group, there was no correlation between body mass index (BMI) and TCF7L2 expression. In morbidly obese nondiabetic patients, there was a positive correlation between TCF7L2 expression and BMI (R(2)=0.21). In morbidly obese diabetic patients, there was an inverse correlation between TCF7L2 expression and BMI (R(2)=0.58). There was no significant relationship between TCF7L2 expression and age or glycosylated hemoglobin (HbA1c). The cause of diabetes is multifactorial but the data from our pilot study documents the relationship of TCF7L2 with type 2 diabetes in morbidly obese patients.

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.

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

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.

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

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...

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.

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.

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...

Occurrence and quantification of Shiga toxin-producing Escherichia coli from food matrices

Directory of Open Access Journals (Sweden)

C. Sethulekshmi

2018-02-01

Full Text Available Aim: The objective of the study was to detect Shiga toxin-producing Escherichia coli (STEC and develop a quantitative polymerase chain reaction (qPCR assay to quantify the bacterial DNA present in different food matrices. Materials and Methods: A total of 758 samples were collected during a period from January 2015 to December 2016 from Kozhikode, Thrissur, and Alappuzha districts of Kerala. The samples consisted of raw milk (135, pasteurized milk (100, beef (132, buffalo meat (130, chevon (104, beef kheema (115, and beef sausage (42. All the samples collected were subjected to isolation and identification of STEC by conventional culture technique. Confirmation of virulence genes was carried out using PCR. For the quantification of STEC in different food matrices, a qPCR was standardized against stx1 gene of STEC by the construction of standard curve using SYBR green chemistry. Results: The overall occurrence of STEC in raw milk (n=135, beef (n=132, buffalo meat (n=130, chevon (n=104, and beef kheema (n=115 samples collected from Kozhikode, Thrissur, and Alappuzha districts of Kerala was 19.26%, 41.6%, 16.92%, 28.85%, and 41.74%, respectively. PCR revealed the presence of stx1 and stx2 genes in 88.46 and 83.64 and 30.77 and 40.00% of STEC isolates from raw milk and beef samples, respectively, while 100% of the STEC isolates from buffalo beef and beef kheema samples carried stx1 gene. Real-time qPCR assay was used to quantify the bacterial cells present in different food matrices. The standard curve was developed, and the slopes, intercept, and R2 of linear regression curves were -3.10, 34.24, and 0.99, respectively. Conclusion: The considerably high occurrence of STEC in the study confirms the importance of foods of animal origin as a vehicle of infection to humans. In the present study, on comparing the overall occurrence of STEC, the highest percentage of occurrence was reported in beef kheema samples. The study shows the need for rigid food

Using multiple PCR and CE with chemiluminescence detection for simultaneous qualitative and quantitative analysis of genetically modified organism.

Science.gov (United States)

Guo, Longhua; Qiu, Bin; Chi, Yuwu; Chen, Guonan

2008-09-01

In this paper, an ultrasensitive CE-CL detection system coupled with a novel double-on-column coaxial flow detection interface was developed for the detection of PCR products. A reliable procedure based on this system had been demonstrated for qualitative and quantitative analysis of genetically modified organism-the detection of Roundup Ready Soy (RRS) samples was presented as an example. The promoter, terminator, function and two reference genes of RRS were amplified with multiplex PCR simultaneously. After that, the multiplex PCR products were labeled with acridinium ester at the 5'-terminal through an amino modification and then analyzed by the proposed CE-CL system. Reproducibility of analysis times and peak heights for the CE-CL analysis were determined to be better than 0.91 and 3.07% (RSD, n=15), respectively, for three consecutive days. It was shown that this method could accurately and qualitatively detect RRS standards and the simulative samples. The evaluation in terms of quantitative analysis of RRS provided by this new method was confirmed by comparing our assay results with those of the standard real-time quantitative PCR (RT-QPCR) using SYBR Green I dyes. The results showed a good coherence between the two methods. This approach demonstrated the possibility for accurate qualitative and quantitative detection of GM plants in a single run.

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.

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...

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.

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...

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.

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.

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...

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.

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.

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.

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...

Relative sensitivity of conventional and real-time PCR assays for detection of SFG Rickettsia in blood and tissue samples from laboratory animals.

Science.gov (United States)

Zemtsova, Galina E; Montgomery, Merrill; Levin, Michael L

2015-01-01

Studies on the natural transmission cycles of zoonotic pathogens and the reservoir competence of vertebrate hosts require methods for reliable diagnosis of infection in wild and laboratory animals. Several PCR-based applications have been developed for detection of infections caused by Spotted Fever group Rickettsia spp. in a variety of animal tissues. These assays are being widely used by researchers, but they differ in their sensitivity and reliability. We compared the sensitivity of five previously published conventional PCR assays and one SYBR green-based real-time PCR assay for the detection of rickettsial DNA in blood and tissue samples from Rickettsia- infected laboratory animals (n = 87). The real-time PCR, which detected rickettsial DNA in 37.9% of samples, was the most sensitive. The next best were the semi-nested ompA assay and rpoB conventional PCR, which detected as positive 18.4% and 14.9% samples respectively. Conventional assays targeting ompB, gltA and hrtA genes have been the least sensitive. Therefore, we recommend the SYBR green-based real-time PCR as a tool for the detection of rickettsial DNA in animal samples due to its higher sensitivity when compared to more traditional assays.

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.

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

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...

Detection of telomerase activity in Plasmodium falciparum using a nonradioactive method

Directory of Open Access Journals (Sweden)

Rubiano Claudia C

2003-01-01

Full Text Available A simple, quick and sensitive method was used to detect telomerase activity in Plasmodium falciparum. The telomeric repeat amplification protocol (TRAP assay was modified using electrophoresis and staining with SYBR-green I to detect telomerase activity in a range of 10² to 10(7 parasites. This might be a useful way to ascertain telomerase activity in different types of nontumor cells.

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.

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.

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.

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.

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...

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...

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...

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.

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.

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

Rapid Identification of Key Pathogens in Wound Infection by Molecular Means

Science.gov (United States)

2006-01-01

3 3 5 4 7 T I NT NT NT NT - - - Prevotella denticola ATCC 33185 1 NT NT NT NT - - - Prevotella intermedia ATCC 2561 IT I NT NT NT NT - - Prevotella...TaqMan and SYBR Green for Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia , tetQ gene and total bacteria...detection of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis. J. Clin.Microbiol. 41, 863-866. (2003). 20. Higgins ,D. et al. CLUSTAL W

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.

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

Bovine Vaccinia in dairy cattle and suspicion of vesicular disease on milkers in Brazil

Directory of Open Access Journals (Sweden)

Thaís Garcia da Silva

2018-05-01

Full Text Available ABSTRACT: Bovine vaccinia (BV is a vesicular disease induced by the Vaccinia virus (VACV that affects milk production and is an occupational zoonosis. This research had the following objectives: (i detection of VACV by qPCR in cattle with clinical suspicion of vesicular disease; (ii symptoms characterization in animals and milkers with clinical suspicion of the disease and virus detection in humans; and (iii identification of risk factors for infections of VACV in herds from several Brazilian states. A total of 471 bovine epithelial samples from dairy farms, in 15 Brazilian states, were evaluated between 2007 and 2012. The samples were tested by quantitative PCR (qPCR using SYBR Green® reagents, validated with a lower limit of detection of 100 TCID50/50µL (1.7x100 viral particles, and 45.1% of VACV positive samples were detected. Using official forms for epidemiological investigation (FORM-IN, the risk factors for VACV infections in cattle were determined to be farms with a lack of technological facilities (P=0.029 and the presence of rodents (P=0.001. There was an effect of seasonality in cattle with a higher occurrence of BV during the dry season. A total of 420 epidemiological questionnaires were applied at public health care centers, where 100% of the milkers had vesicular lesions on their hands (98.1% and on their arms (6.9%. The most frequent clinical symptoms in humans were: local swelling (74.2%, headache (20.7%, fever (10.4% and inguinal lymphadenopathy (74.2%. Only 19.98% of milkers aged between 39 and 58 years were seroreactive to VACV and were immunized with the human anti-smallpox vaccine. There was an increase in the frequency of BV in older individuals due to their natural decrease in specific immunity. It has been shown that the implementation of zootechnical management techniques and health planning are important for the prevention of BV in animals and humans.

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.

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

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...

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.

Target-induced structure switching of hairpin aptamers for label-free and sensitive fluorescent detection of ATP via exonuclease-catalyzed target recycling amplification.

Science.gov (United States)

Xu, Yunying; Xu, Jin; Xiang, Yun; Yuan, Ruo; Chai, Yaqin

2014-01-15

In this work, we described the development of a new label-free, simple and sensitive fluorescent ATP sensing platform based on exonuclease III (Exo III)-catalyzed target recycling (ECTR) amplification and SYBR Green I indicator. The hairpin aptamer probes underwent conformational structure switching and re-configuration in the presence of ATP, which led to catalytic cleavage of the re-configured aptamers by Exo III to release ATP and to initiate the ECTR process. Such ECTR process resulted in the digestion of a significant number of the hairpin aptamer probes, leading to much less intercalation of SYBR Green I to the hairpin stems and drastic suppression of the fluorescence emission for sensitive ATP detection down to the low nanomolar level. Due to the highly specific affinity bindings between aptamers and ATP, the developed method exhibited excellent selectivity toward ATP against other analogous molecules. Besides, our ATP sensing approach used un-modified aptamer probes and could be performed in a "mix-and-detect" fashion in homogenous solutions. All these distinct advantages of the developed method thus made it hold great potential for the development of simple and robust sensing strategies for the detection of other small molecules. © 2013 Elsevier B.V. All rights reserved.

Relative sensitivity of conventional and real-time PCR assays for detection of SFG Rickettsia in blood and tissue samples from laboratory animals.

Directory of Open Access Journals (Sweden)

Galina E Zemtsova

Full Text Available Studies on the natural transmission cycles of zoonotic pathogens and the reservoir competence of vertebrate hosts require methods for reliable diagnosis of infection in wild and laboratory animals. Several PCR-based applications have been developed for detection of infections caused by Spotted Fever group Rickettsia spp. in a variety of animal tissues. These assays are being widely used by researchers, but they differ in their sensitivity and reliability. We compared the sensitivity of five previously published conventional PCR assays and one SYBR green-based real-time PCR assay for the detection of rickettsial DNA in blood and tissue samples from Rickettsia- infected laboratory animals (n = 87. The real-time PCR, which detected rickettsial DNA in 37.9% of samples, was the most sensitive. The next best were the semi-nested ompA assay and rpoB conventional PCR, which detected as positive 18.4% and 14.9% samples respectively. Conventional assays targeting ompB, gltA and hrtA genes have been the least sensitive. Therefore, we recommend the SYBR green-based real-time PCR as a tool for the detection of rickettsial DNA in animal samples due to its higher sensitivity when compared to more traditional assays.

Evaluation of a real-time PCR assay based on the single-copy SAG1 gene for the detection of Toxoplasma gondii.

Science.gov (United States)

Yu, Haijie; Huang, Bin; Zhuo, Xunhui; Chen, Xueqiu; Du, Aifang

2013-11-08

Real-time PCR-based detection of Toxoplasma gondii is very sensitive and convenient for diagnosing toxoplasmosis. However, the performance of the PCR assays could be influenced by the target gene chosen. Here we evaluate a real-time PCR assay using double-stranded DNA dyes (SYBR(®) Green I assay) with a new set of primers targeting the SAG1 gene for the fast and specific detection of T. gondii. The assay showed higher sensitivity than conventional PCR protocols using T. gondii DNA as template. The detection limit of the developed real-time PCR assay was in the order of 1 tachyzoite. The assay was also assessed by experimentally infected mice and showed positive results for blood (25%), spleen (50%) and lung (50%) as early as 1 dpi. The specificity of the assay was confirmed by using DNA from Neospora caninum, Escherichia coli, Babesia bovis, Trypanosoma brucei, Cryptosporidium parvum, and Toxocara canis. Assay applicability was successfully tested in blood samples collected from slaughtered pigs. These results indicate that, based on SYBR(®) green I, the quantitative SAG1 assay may also be useful in the study of the pathogenicity, immunoprophylaxis, and treatment of T. gondii. Copyright © 2013 Elsevier B.V. All rights reserved.

Endpoint visual detection of three genetically modified rice events by loop-mediated isothermal amplification.

Science.gov (United States)

Chen, Xiaoyun; Wang, Xiaofu; Jin, Nuo; Zhou, Yu; Huang, Sainan; Miao, Qingmei; Zhu, Qing; Xu, Junfeng

2012-11-07

Genetically modified (GM) rice KMD1, TT51-1, and KF6 are three of the most well known transgenic Bt rice lines in China. A rapid and sensitive molecular assay for risk assessment of GM rice is needed. Polymerase chain reaction (PCR), currently the most common method for detecting genetically modified organisms, requires temperature cycling and relatively complex procedures. Here we developed a visual and rapid loop-mediated isothermal amplification (LAMP) method to amplify three GM rice event-specific junction sequences. Target DNA was amplified and visualized by two indicators (SYBR green or hydroxy naphthol blue [HNB]) within 60 min at an isothermal temperature of 63 °C. Different kinds of plants were selected to ensure the specificity of detection and the results of the non-target samples were negative, indicating that the primer sets for the three GM rice varieties had good levels of specificity. The sensitivity of LAMP, with detection limits at low concentration levels (0.01%−0.005% GM), was 10- to 100-fold greater than that of conventional PCR. Additionally, the LAMP assay coupled with an indicator (SYBR green or HNB) facilitated analysis. These findings revealed that the rapid detection method was suitable as a simple field-based test to determine the status of GM crops.

Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification

Directory of Open Access Journals (Sweden)

Qing Zhu

2012-11-01

Full Text Available Genetically modified (GM rice KMD1, TT51-1, and KF6 are three of the most well known transgenic Bt rice lines in China. A rapid and sensitive molecular assay for risk assessment of GM rice is needed. Polymerase chain reaction (PCR, currently the most common method for detecting genetically modified organisms, requires temperature cycling and relatively complex procedures. Here we developed a visual and rapid loop-mediated isothermal amplification (LAMP method to amplify three GM rice event-specific junction sequences. Target DNA was amplified and visualized by two indicators (SYBR green or hydroxy naphthol blue [HNB] within 60 min at an isothermal temperature of 63 °C. Different kinds of plants were selected to ensure the specificity of detection and the results of the non-target samples were negative, indicating that the primer sets for the three GM rice varieties had good levels of specificity. The sensitivity of LAMP, with detection limits at low concentration levels (0.01%–0.005% GM, was 10- to 100-fold greater than that of conventional PCR. Additionally, the LAMP assay coupled with an indicator (SYBR green or HNB facilitated analysis. These findings revealed that the rapid detection method was suitable as a simple field-based test to determine the status of GM crops.

Aptamer-fluorescent silica nanoparticles bioconjugates based dual-color flow cytometry for specific detection of Staphylococcus aureus.

Science.gov (United States)

He, Xiaoxiao; Li, Yuhong; He, Dinggen; Wang, Kemin; Shangguan, Jingfang; Shi, Hui

2014-07-01

This paper describes a sensitive and specific determination strategy for Staphylococcus aureus (S. aureus) detection using aptamer recognition and fluorescent silica nanoparticles (FSiNPs) label based dual-color flow cytometry assay (Aptamer/FSiNPs-DCFCM). In the protocol, an aptamer, having high affinity to S. aureus, was first covalently immobilized onto chloropropyl functionalized FSiNPs through a click chemistry approach to generate aptamer-nanoparticles bioconjugates (Aptamer/FSiNPs). Next, S. aureus was incubated with Aptamer/FSiNPs, and then stained with SYBR Green I (a special staining material for the duplex DNA). Upon target binding and nucleic acid staining with SYBR Green I, the S. aureus was determined using two-color flow cytometry. The method took advantage of the specificity of aptamer, signal amplification of FSiNPs label and decreased false positives of two-color flow cytometry assay. It was demonstrated that these Aptamer/FSiNPs could efficiently recognize and fluorescently label target S. aureus. Through multiparameter determination with flow cytometry, this assay allowed for detection of as low as 1.5 x 10(2) and 7.6 x 10(2) cells mL(-1) S. aureus in buffer and spiked milk, respectively, with higher sensitivity than the Aptamer/FITC based flow cytometry.

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

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.

Occurrence of four species of algae in the marine water of Hong Kong.

Science.gov (United States)

Chai, Yemao; Deng, Wen-Jing; Qin, Xing; Xu, Xiangrong

2017-11-30

Harmful algal blooms (HABs) have broken out frequently throughout the world in recent decades; they are caused by the rapid multiplication of algal cells in near-coastal waters polluted with nitrogen and phosphorus and greatly affect the quality of marine water and human health. Over the past several decades, climate change and increasing environmental degradation have provided favourable growth conditions for certain phytoplankton species. Therefore, it is essential to rapidly identify and enumerate harmful marine algae to control these species. In this study, quantitative PCR (qPCR) was used to detect four representative species of HABs that are widespread in the marine water of Hong Kong, namely, Alexandrium catenella, Pseudo-nitzschia spp., Karenia mikimotoi and Heterosigma akashiwo. We applied qPCR with the dye SYBR Green to detect Alexandrium spp. and Pseudo-nitzschia spp. and used TaqMan probe for the enumeration of Karenia mikimotoi and Heterosigma akashiwo. The total genomic DNA of these algae from Hong Kong marine water was extracted successfully using the CTAB method, and for each kind of alga, we constructed a ten-fold series of recombinant plasmid solutions containing certain gene fragments of 18S rDNA and ITS1-5.8S-ITS2 as standard samples. Ten-fold dilutions of the DNA of known numbers of the extracted algal cells were also used to create an additional standard curve. In this way, the relationship between the cell number and the related plasmid copy number was established. The qPCR assay displayed high sensitivity in monitoring marine water samples in which the low concentrations of harmful algae were not detected accurately by traditional methods. The results showed that the cell numbers of the four species were all in low abundance. For Alexandrium catenella, the cell abundances at 12 sites ranged from 3.8×10 2 to 4.3×10 3 cellsL -1 , while H. akashiwo, K. mikimotoi and Pseudo-nitzschia ranged from 1.1×10 2 to 1.3×10 3 , from 23 to 6.5×10 2

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.

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.

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.

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...

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.

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

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.

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.

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...

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...

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 ...

Real-time PCR-based genotyping from whole blood using Taq DNA polymerase and a buffer supplemented with 1,2-propanediol and trehalose

Czech Academy of Sciences Publication Activity Database

Utekal, Pavol; Kocanda, Lukáš; Matoušek, P.; Wagner, P.; Bugajev, Viktor; Dráber, Petr

2015-01-01

Roč. 416, January (2015), s. 178-182 ISSN 0022-1759 R&D Projects: GA ČR(CZ) GBP302/12/G101; GA MPO FR-TI3/067; GA ČR(CZ) GA14-09807S; GA ČR(CZ) GA14-00703S Institutional support: RVO:68378050 Keywords : 1,2-Propanediol * real-time PCR * SYBR Green I * Taq DNA polymerase * trehalose * Unseparated blood Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.858, year: 2015

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.

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...

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.

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...

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

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 ...

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

[Application of rapid PCR to authenticate medicinal snakes].

Science.gov (United States)

Chen, Kang; Jiang, Chao; Yuan, Yuan; Huang, Lu-Qi; Li, Man

2014-10-01

To obtained an accurate, rapid and efficient method for authenticate medicinal snakes listed in Chinese Pharmacopoeia (Zaocysd humnades, Bungarus multicinctus, Agkistrodon acutus), a rapid PCR method for authenticate snakes and its adulterants was established based on the classic molecular authentication methods. DNA was extracted by alkaline lysis and the specific primers were amplified by two-steps PCR amplification method. The denatured and annealing temperature and cycle numbers were optimized. When 100 x SYBR Green I was added in the PCR product, strong green fluorescence was visualized under 365 nm UV whereas adulterants without. The whole process can complete in 30-45 minutes. The established method provides the technical support for authentication of the snakes on field.

Green power programs in Canada : 2002 : Overview of Government green power policies, utility green power development programs, green power and certificate marketing initiatives, and their benefits

International Nuclear Information System (INIS)

Bramley, M.; Boustie, S.; Vadgama, J.; Wieler, C.; Pape-Salmon, A.; Holmes, R.

2003-11-01

Green power is generally 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. Green power 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 associated with these four categories in 2002 were discussed in this report. However, difficulties with quantification prevented the inclusion of some green power activities in the report, 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. Each category was presented in detail. The information included in the report was based on surveys sent to each program proponent. Follow-up communications and other publicly available information was also included. New programs operating in 2003 or currently under development were listed. refs., 8 tabs

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.

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...

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...

Tracking vaginal, anal and oral infection in a mouse papillomavirus infection model.

Science.gov (United States)

Hu, Jiafen; Budgeon, Lynn R; Cladel, Nancy M; Balogh, Karla; Myers, Roland; Cooper, Timothy K; Christensen, Neil D

2015-12-01

Noninvasive and practical techniques to longitudinally track viral infection are sought after in clinical practice. We report a proof-of-principle study to monitor the viral DNA copy number using a newly established mouse papillomavirus (MmuPV1) mucosal infection model. We hypothesized that viral presence could be identified and quantified by collecting lavage samples from cervicovaginal, anal and oral sites. Nude mice infected at these sites with infectious MmuPV1 were tracked for up to 23 weeks starting at 6 weeks post-infection. Viral DNA copy number was determined by SYBR Green Q-PCR analysis. In addition, we tracked viral DNA load through three complete oestrous cycles to pinpoint whether there was a correlation between the DNA load and the four stages of the oestrous cycle. Our results showed that high viral DNA copy number was reproducibly detected from both anal and cervicovaginal lavage samples. The infection and disease progression were further confirmed by histology, cytology, in situ hybridization, immunohistochemistry and transmission electron microscopy. Interestingly, the viral copy number fluctuated over the oestrous cycle, with the highest level at the oestrus stage, implying that multiple sampling might be necessary to provide a reliable diagnosis. Virus DNA was detected in oral lavage samples at a later time after infection. Lower viral DNA load was found in oral samples when compared with those in anal and vaginal tracts. To our knowledge, our study is the first in vivo study to sequentially monitor papillomavirus infection from mucosal anal, oral and vaginal tracts in a preclinical model.

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.

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...

Development of a Real-time PCR test for porcine group A rotavirus diagnosis

OpenAIRE

Marconi, Elizabeth C.M.; Bernardes, Nara T.C.G.; Beserra, Laila A.R.; Silva, Fernanda D.F.; Gregori, Fabio

2015-01-01

Group A Rotavirus (RVA) is one of the most common causes of diarrhea in humans and several animal species. A SYBR-Green Real-Time polymerase chain reaction (PCR) was developed to diagnose RVA from porcine fecal samples, targeting amplification of a 137-bp fragment of nonstructural protein 5 (NSP5) gene using mRNA of bovine NADH-desidrogenase-5 as exogenous internal control. Sixty-five samples were tested (25 tested positive for conventional PCR and genetic sequencing). The overall agreement (...

The Influence of Green Marketing on Green Satisfaction Mediated By Perceived Quality and Its Impact to Green Trust in Injection Motorcycle

Directory of Open Access Journals (Sweden)

Shelvy Kurniawan

2014-09-01

Full Text Available Currently, motorcycle manufacturers are increasingly motivated to replace their motorcycle into fuel injection products. The growing concern from the consumers to the environment and the regulations of emission standards, that is Euro 3, for motorcycle industry is being finalized in the Ministry of Environment in order to be implemented in Indonesia. Through this research, the writer will examine the effect of green marketing on perceived quality, green satisfaction, and green trust, the effect of perceived quality on green satisfaction, and the effect of green satisfaction on green trust. Those effects needs to be investigated in order to know how far the effects of green marketing and to ensure whether green marketing is well accepted or not by the market in motorcycle industry. Scope of this research is also limited to the user of fuel injection motorcycle in Jakarta for Honda and Yamaha who involved as decision maker when the motorcycle is purchased. Sampling technique used in this research is quota sampling and the analysis method is structural equation modeling (SEM. The findings of this research are: green marketing has a significant direct effect on perceived quality, perceived quality has a significant direct effect on green satisfaction, green satisfaction has a significant direct effect on green trust, green marketing has a significant direct and indirect effect on green satisfaction, and green marketing has a significant direct and indirect effect on green trust. All of those effects are found to be positive effects.

Purchasing green to become greener: Factors influence consumers’ green purchasing behavior

Directory of Open Access Journals (Sweden)

Hosein Vazifehdoust

2013-09-01

Full Text Available This study proposes an integrated model that combines the Theory of Reasoned Action (TRA and two categories of variables, personal and marketing, to investigate the attitudinal and behavioral decision factors to purchase green products. The model derived and tested via structural equation modeling on a sample of 374 consumers from the Guilan province in Iran. The results show that attitude is explained by consumers’ environmental concern, quality of green products, green advertising and green labeling. The results of the structural equation analysis indicate that attitude positively influences intention to purchase green products. Green purchasing intention also influences on green purchasing behavior. This paper also discusses the implications of the results for marketers and researchers.

Is 'green finance' actually green? 'Make The Planet Great Again' or 'Green-washing', we must choose. Report on green bonds and climate bonds

International Nuclear Information System (INIS)

Combes, Maxime; Plihon, Dominique; Zippert, Jean-Sebastien; Chaussalet, Alexis; Planche, Jeanne; Poulain, Melanie

2017-12-01

As Paris dreams of becoming the capital of green finance, the author proposes a discussion of the emerging market of green bonds, and formulates a set of recommendations for this new financial instrument not to be polluted by green-washing operations. He first describes what a green bond is, and then comments what the green bond market represents, discusses development perspectives for this market, comments the Paris dream of becoming the world capital of a green and sustainable finance. He explains why this green bond market appears to be so interesting, and what guarantees that a green bond will finance green projects. He discusses the role of rating agencies, whether the emitter of a green bond must be green, and the impact of green bonds on climate, on the environment and on populations. He discusses the possible evolution towards a constraining regulation, and examines whether this system can be an operational financing source for energy transition. Recommendations concern the market regulation by public authorities, the creation of a new rating agency for green finance, how to make the world bond market climate-compatible, and the creation of other financing channels for actors who have no access to the bond market

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...

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...

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.

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...

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.

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.

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.

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.

Green Roofs: Standardization and Quality Control of Processes in Green Construction

Directory of Open Access Journals (Sweden)

Korol Elena

2017-01-01

Full Text Available The article considers the problems of standardization and quality control of processes in the construction, improvement of integrated safety of buildings and the implementation of innovative green building technologies, the use of national standards as well as international rating systems for green buildings evaluation. This is one of the priority directions in development of the modern construction. The aim of this study is the analysis of the green roof systems and international standards, which were carried out in the green building industry. The authors have studied traditional and innovative solutions of rational using natural resources and energy, the green roof system with integration of supported solar and wind energy collecting and converting devices and of irrigation system. Some studies provide evidence for the benefits of the modular green roof system in urban green space with microclimate differences. This article presents a new research which advances our knowledge of the economic and environmental services provided by the green roof system. Research reported here also considers the analysis of the Russian and international legislation of the quality control of processes in green construction.

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)

The synergy in green persuasion: Green celebrity endorsers in green advertising: A study of brand-endorser congruence effects in green advertising

OpenAIRE

Blasche, J.; Ketelaar, P.E.

2015-01-01

This study examines celebrity endorser-brand congruence effects in green advertising on the ads' effectiveness. In an experimental survey, Dutch participants (197) saw ads with a congruent or incongruent celebrity endorser. Extending the match-up hypothesis to a novel match-up factor, greenness, the results show that pro-environmental celebrity endorsers yield more favourable attitudes towards the ad, the brand, and purchase intentions compared to non-green celebrity endorsers. Practitioners ...

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...

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...

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.

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.

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...

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.

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.

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.

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)

Analytical Methods for Malachite Green : Completion Report : Malachite Green Analysis in Water.

Energy Technology Data Exchange (ETDEWEB)

Allen, John L.; Gofus, Jane E.; Meinertz, Jeffery R.

1991-06-01

Malachite green is a known teratogen and therefore its use is limited to nonfood fish under an Investigational New Animal Drug permit (INAD), number 2573. Although a charcoal adsorption column was developed to remove malachite green from hatchery water, INAD compliance requires that the malachite green residue concentrations in any effluent from hatcheries using the chemical be quantified. Therefore, we developed a method for the analysis of malachite green residues in water. Enrichment of the residues of malachite green in water on a diol column followed by High Performance Liquid Chromatographic (HPLC) analysis gives a minimum sensitivity of less than 10 ppb for the chemical. When combined with post-column oxidation using a lead oxide post-column reactor, the procedure can be used for the simultaneous analysis of malachite green in its leuco form, a decomposition product of the dye, as well as its chromatic form. Recovery of the leuco form is pH dependent and water samples should be adjusted to pH 6 to optimize recovery of this form. Water samples spiked with malachite green were concentrated on a diol column followed by elution with 0.05 M p-toluene sulfonic acid in methanol. The methanol elutes were analyzed by HPLC. Pond water samples spiked with malachite green and leuco malachite green yielded average recoveries of 95.4% for malachite green and 57.3% for leuco malachite green. Tap water samples spiked with the carbinol form of malachite green gave average recoveries of 98.6%. The method is very sensitive and is capable of detecting malachite green residues in water at less than 10 ppb. Fish culturists, who cannot find an effective replacement for malachite green, can utilize the method to ensure that their effluents comply with INAD regulations. 13 refs., 2 figs., 7 tabs.

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...

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.

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 ...

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.

Detection of mycoplasmas in goat milk by flow cytometry.

Science.gov (United States)

Assunção, Patricia; Davey, Hazel M; Rosales, Ruben S; Antunes, Nuno T; de la Fe, Christian; Ramirez, Ana S; de Galarreta, Carlos M Ruiz; Poveda, Jose B

2007-12-01

The detection of mycoplasma in milk can be performed by either culture techniques or polymerase chain reaction (PCR) based methods. Although PCR can reduce the average diagnostic time to 5 h in comparison with the several days for the isolation of the agent, there is still a need to develop methods, which could give earlier results. For this purpose, we tested the ability of flow cytometry (FC) to detect mycoplasmas in milk samples. Milk samples inoculated with four different mycoplasmas, Mycoplasma agalactiae, Mycoplasma putrefaciens, Mycoplasma capricolum subsp. Capricolum, or Mycoplasma mycoides subsp. mycoides large-colony type, known to cause contagious agalactia in goats, were stained with the DNA stain SYBR Green I and analyzed by FC. Three goat milk samples, from which mycoplasmas have been isolated in broth medium were also analyzed. All mycoplasmas were easily distinguished from debris of milk samples, but it was not possible to distinguish between the different mycoplasma species. In our conditions, the detection limit of the technique was of the order of 10(3)-10(4) cells ml(-1). Furthermore, mycoplasmas were also distinguished from Staphylococcus aureus. FC together with SYBR Green I was able to distinguish between mycoplasma cells and debris present in milk samples and gave results in 20-30 min. This is an important first step in developing a robust, routine flow cytometric method for the detection of mycoplasmas in milk samples. (c) 2007 International Society for Analytical Cytology

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.

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

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...

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)

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

Association between periodontal condition and subgingival microbiota in women during pregnancy: a longitudinal study.

Science.gov (United States)

Borgo, Priscila Viola; Rodrigues, Viviane Aparecida Arenas; Feitosa, Alfredo Carlos Rodrigues; Xavier, Karla Correa Barcelos; Avila-Campos, Mario Julio

2014-01-01

In this study, the gingival conditions and the quantitative detection for Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia in pregnant women were determined. Quantitative determinations of periodontal bacteria by using a SyBr green system in women during pregnancy were performed. Women at the 2nd and 3rd trimesters of pregnancy and non-pregnant women were included in this study. A. actinomycetemcomitans was observed in high numbers in women at the 2nd and 3rd trimesters of pregnancy with a significant difference (pperiodontal disease.

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.

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.

Green Roofs: A Part of Green Infrastructure Strategy for Urban Areas

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 provides insight for the end user as to the benefits for green roof technology. It provides links to currently available EPA rep...

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

The synergy in green persuasion: Green celebrity endorsers in green advertising: A study of brand-endorser congruence effects in green advertising

NARCIS (Netherlands)

Blasche, J.; Ketelaar, P.E.

2015-01-01

This study examines celebrity endorser-brand congruence effects in green advertising on the ads' effectiveness. In an experimental survey, Dutch participants (197) saw ads with a congruent or incongruent celebrity endorser. Extending the match-up hypothesis to a novel match-up factor, greenness, the

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.

Greening Steel Work: Varieties of Capitalism and the "Greening" of Skills

Science.gov (United States)

Evans, Claire; Stroud, Dean

2016-01-01

An important driver of change in work, employment and skills is European Union policy aims of sustainable economic growth and the cultivation of a green economy. Part of the latter--which is supported by increasing environmental regulation--focuses on the development of a "green skills agenda," which involves the "greening" of…

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…

The green dilemma: Reflections of a Generation Y consumer cohort on green purchase behaviour

Directory of Open Access Journals (Sweden)

A. Muposhi

2015-12-01

Full Text Available Green consumerism has garnered much scholarly interest in recent years. However, research on the influence of the Social Dilemma Theory (SDT on green purchase behaviour has been scarce. Using data generated from sixteen in-depth-interviews, the present study identified perceived efficacy, perceived cost, in-group and self-identity, trust and peer influence as the main antecedents of SDT that influence green purchase behaviour. The findings of the study imply that to promote and institutionalise green purchase behaviour, marketers need to enhance perceived efficacy, trust in green products, reduce perceived cost, align green products with the consumers’ sought image and utilise peer networks when structuring green marketing messages.

The Prevalence of Trypanosoma cruzi, the Causal Agent of Chagas Disease, in Texas Rodent Populations.

Science.gov (United States)

Aleman, Adriana; Guerra, Trina; Maikis, Troy J; Milholland, Matthew T; Castro-Arellano, Ivan; Forstner, Michael R J; Hahn, Dittmar

2017-03-01

Rodent species were assessed as potential hosts of Trypanosoma cruzi, the etiologic agent of Chagas disease, from five sites throughout Texas in sylvan and disturbed habitats. A total of 592 rodents were captured, resulting in a wide taxonomic representation of 11 genera and 15 species. Heart samples of 543 individuals were successfully analyzed by SybrGreen-based quantitative PCR (qPCR) targeting a 166 bp fragment of satellite DNA of T. cruzi. Eight rodents representing six species from six genera and two families were infected with T. cruzi. This is the first report of T. cruzi in the pygmy mouse (Baiomys taylori) and the white-footed mouse (Peromyscus leucopus) for the USA. All infected rodents were from the southernmost site (Las Palomas Wildlife Management Area). No differences in pathogen prevalence existed between disturbed habitats (5 of 131 tested; 3.8%) and sylvan habitats (3 of 40 tested; 7.5%). Most positives (n = 6, 16% prevalence) were detected in late winter with single positives in both spring (3% prevalence) and fall (1% prevalence). Additionally, 30 Triatoma insects were collected opportunistically from sites in central Texas. Fifty percent of these insects, i.e., 13 T. gerstaeckeri (68%), and two T. lecticularia (100%) were positive for T. cruzi. Comparative sequence analyses of 18S rRNA of samples provided identical results with respect to detection of the presence or absence of T. cruzi and assigned T. cruzi from rodents collected in late winter to lineage TcI. T. cruzi from Triatoma sp. and rodents from subsequent collections in spring and fall were different, however, and could not be assigned to other lineages with certainty.

Is green economy achievable through championing green growth? A local government experience from Zambia

Directory of Open Access Journals (Sweden)

Phiri Rodgers

2016-03-01

Full Text Available The need to enhance environmental sustainability, sustainable development and growth that takes into account the well-being of the people and nature because of the increased production and consumption of goods and services is the major driver to the introduction of green economy in Zambia and countries in southern Africa. This article examines the extent to which local government in Zambia has embraced green growth and green economy and critically analyses the concept of green economy and green growth. This study is based on a review of planning and policy documents, a household questionnaire survey and interviews with various institutions, planners and rural development organisations. A number of policies implemented at the local government level were analysed and reflected upon irrespective of whether they contain the components of green growth and green economy and the extent to which they contribute to attaining green economy. The article argues that the need for economic diversification is important as far as green economy is concerned. The article recommends the need to invest in research and development in order to find more carbon-free economic activities. The conclusion is that local government is key to achieving green growth and green economy, because it is involved at all levels, from policy formulation to implementation.

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/

Relationship between green marketing strategies and green marketing credibility among Generation Y

OpenAIRE

Garcia Sandoval, Michelle Haeberli; Manon Padilla, Alejandro

2016-01-01

Since terms like “sustainability” and “consumer consciousness” were introduced, green products began being integrated into consumers’ lifestyles. But due to the greenwashing practices that took place during the 90’s consumers refrain to buy green products because they do not trust the advertising released by marketers. The purpose of this thesis is to explore the relationship between green advertising credibility (dependent variable) and price sensitiveness, and the four proposed green market...

Green Roofs and Green Walls for Biodiversity Conservation: A Contribution to Urban Connectivity?

Directory of Open Access Journals (Sweden)

Flavie Mayrand

2018-03-01

Full Text Available Green roofs and walls have recently emerged as conservation tools, and they offer promising additional opportunities to enhance biodiversity in cities. However, their ecological conditions remain poorly considered when planning wildlife corridors. To discuss the role of vegetated buildings in landscape connectivity, we reviewed the ecological and technical specificities of green walls and green roofs in light of the key factors concerning urban wildlife (patch size, quality, abundance, and isolation. Green roofs and walls show limited patch sizes, distinct habitat quality at the building scale, and limited redundancy of patch quality within the landscape. We also highlight that the abundance of roof and wall patches is often low. Future research is needed to establish if walls can be vertical corridors for wildlife, thereby reducing the isolation of green roofs. We argue that creating 3D ecological connectivity within the city requires substantial modifications of the design and maintenance of existing green building systems. We suggest that research is needed to integrate the biotic and abiotic characteristics of green buildings to make them more closely resemble those of open green spaces.

Modeling the NPE with finite sources and empirical Green`s functions

Energy Technology Data Exchange (ETDEWEB)

Hutchings, L.; Kasameyer, P.; Goldstein, P. [Lawrence Livermore National Lab., CA (United States)] [and others

1994-12-31

In order to better understand the source characteristics of both nuclear and chemical explosions for purposes of discrimination, we have modeled the NPE chemical explosion as a finite source and with empirical Green`s functions. Seismograms are synthesized at four sties to test the validity of source models. We use a smaller chemical explosion detonated in the vicinity of the working point to obtain empirical Green`s functions. Empirical Green`s functions contain all the linear information of the geology along the propagation path and recording site, which are identical for chemical or nuclear explosions, and therefore reduce the variability in modeling the source of the larger event. We further constrain the solution to have the overall source duration obtained from point-source deconvolution results. In modeling the source, we consider both an elastic source on a spherical surface and an inelastic expanding spherical volume source. We found that the spherical volume solution provides better fits to observed seismograms. The potential to identify secondary sources was examined, but the resolution is too poor to be definitive.

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

Green Campus initiative and its impacts on quality of life of stakeholders in Green and Non-Green Campus universities.

Science.gov (United States)

Tiyarattanachai, Ronnachai; Hollmann, Nicholas M

2016-01-01

In 2010, Universitas Indonesia (UI) developed the UI GreenMetric World University Ranking for universities to share information about their sustainability practices. This ranking system was well aligned with the basis of Sustainability for Higher Education. The scoring system can also be used as a guideline for universities to achieve sustainability in their campuses. Since its first launch, more universities around the world have increasingly participated in the ranking system including many universities in Thailand. This study compared perception of stakeholders in Green Campus and Non-Green Campus universities in Thailand regarding stakeholders' satisfaction on sustainability practices and perceived quality of life at their campuses. The results showed that stakeholders at the studied Green Campus University were more satisfied and had significantly better perceived quality of life compared to stakeholders from the studied Non-Green Campus university. The results suggested that universities should adopt the criteria set in the UI GreenMetric World University Ranking to achieve better sustainability in their campuses and improve quality of life of their stakeholders.

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…

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.

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...

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...

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...

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...

GREEN MARKETING –GO GREEN FOR THE SUSTAINABLE DEVELOPMENT OF THE PUBLIC

OpenAIRE

J. Kavitha

2016-01-01

Environment plays an important role in our lives. The Humans are only responsible for the environment. The initiatives should be taken from every individual then the day is not so far when global warming could be controlled. In the phrase “GREEN MARKETING” green signifies eco-friendly innovation. The objective of this study is to examine the growth of green marketing sector & its future. The concept of green marketing is originated primarily in the developed markets and rapidly gaining scope ...

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.

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

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.

Green space definition affects associations of green space with overweight and physical activity.

Science.gov (United States)

Klompmaker, Jochem O; Hoek, Gerard; Bloemsma, Lizan D; Gehring, Ulrike; Strak, Maciej; Wijga, Alet H; van den Brink, Carolien; Brunekreef, Bert; Lebret, Erik; Janssen, Nicole A H

2018-01-01

In epidemiological studies, exposure to green space is inconsistently associated with being overweight and physical activity, possibly because studies differ widely in their definition of green space exposure, inclusion of important confounders, study population and data analysis. We evaluated whether the association of green space with being overweight and physical activity depended upon definition of greenspace. We conducted a cross-sectional study using data from a Dutch national health survey of 387,195 adults. Distance to the nearest park entrance and surrounding green space, based on the Normalized Difference Vegetation Index (NDVI) or a detailed Dutch land-use database (TOP10NL), was calculated for each residential address. We used logistic regression analyses to study the association of green space exposure with being overweight and being moderately or vigorously physically active outdoors at least 150min/week (self-reported). To study the shape of the association, we specified natural splines and quintiles. The distance to the nearest park entrance was not associated with being overweight or outdoor physical activity. Associations of surrounding green space with being overweight or outdoor physical activity were highly non-linear. For NDVI surrounding greenness, we observed significantly decreased odds of being overweight [300m buffer, odds ratio (OR) = 0.88; 95% CI: 0.86, 0.91] and increased odds for outdoor physical activity [300m buffer, OR = 1.14; 95% CI: 1.10, 1.17] in the highest quintile compared to the lowest quintile. For TOP10NL surrounding green space, associations were mostly non-significant. Associations were generally stronger for subjects living in less urban areas and for the smaller buffers. Associations of green space with being overweight and outdoor physical activity differed considerably between different green space definitions. Associations were strongest for NDVI surrounding greenness. Copyright © 2017 The Authors. Published by

City of Austin: Green habitat learning project. A green builder model home project

Energy Technology Data Exchange (ETDEWEB)

NONE

1995-12-01

The purpose of the Year 14 UCETF project was to design and construct a residential structure that could serve as a demonstration facility, training site, and testing and monitoring laboratory for issues related to the implementation of sustainable building practices and materials. The Model Home Project builds on the previous and existing efforts, partially funded by the UCETF, of the City of Austin Green Builder Program to incorporate sustainable building practices into mainstream building activities. The Green Builder Program uses the term {open_quotes}green{close_quotes} as a synonym for sustainability. In the research and analysis that was completed for our earlier reports in Years 12 and 13, we characterized specific elements that we associate with sustainability and, thus, green building. In general, we refer to a modified life cycle assessment to ascertain if {open_quotes}green{close_quotes} building options reflect similar positive cyclical patterns found in nature (i.e. recyclability, recycled content, renewable resources, etc.). We additionally consider economic, human health and synergistic ecological impacts associated with our building choices and characterize the best choices as {open_quotes}green.{close_quotes} Our ultimate goal is to identify and use those {open_quotes}green{close_quotes} materials and processes that provide well for us now and do not compromise similar benefits for future generations. The original partnership developed for this project shifted during the year from a project stressing advanced (many prototypical) {open_quotes}green{close_quotes} building materials and techniques in a research and demonstration context, to off-the-shelf but underutilized {open_quotes}green{close_quotes} materials in the practical social context of using {open_quotes}green{close_quotes} technologies for low income housing. That project, discussed in this report, is called the Green Habitat Learning Project.

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 ...

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.

A market for green certificates may cause less green electricity to be produced

International Nuclear Information System (INIS)

Haugneland, Petter

2004-01-01

The Norwegian government wants to establish in 2006 a market for trading with green certificates which will be issued to producers of new renewable electricity. These certificates will be sold to the consumers, which will be instructed to by a certain amount of green electricity. In 2005 a market will be established for trading with emission quotas of greenhouse gases; in this market, power producers and other industry that emits greenhouse gases must buy emission permits. Some experts, however, say that a market for trading with green certificates may at worst give less production of green electricity, counter to the intention. But a quota system may indirectly increase the production of green electricity, and at the same time one avoids many of the inconveniences involved in a green certificate market

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.

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.

External Audit Green Deal Approach. Final report; Externe Audit Green Deal Aanpak. Eindrapport

Energy Technology Data Exchange (ETDEWEB)

Van Mil, B.P.A.; Gooskens, B.J.F.; Van Schelven, R.M.; Stutje, A.

2013-10-15

External audit of the effectiveness of the Green Deals, based on the question how the Green Deal contributes to green growth. The central idea of this new tool is that the Dutch government facilitates initiatives of businesses, societal organisations, local governments and citizens by removing bottlenecks as much as possible [Dutch] Externe audit over (de effectiviteit van) de Green Deal aanpak, op basis van de volgende onderzoeksvraag: 'Hoe draagt de Green Deal aanpak bij aan het bevorderen van groene groei?' De centrale gedachte van dit nieuwe instrument is dat de overheid initiatieven van bedrijven, maatschappelijke organisaties, decentrale overheden en burgers faciliteert door het wegnemen van knelpunten.

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...

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...

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.

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...

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.

The Ozobranchus leech is a candidate mechanical vector for the fibropapilloma-associated turtle herpesvirus found latently infecting skin tumors on Hawaiian green turtles (Chelonia mydas)

International Nuclear Information System (INIS)

Greenblatt, Rebecca J.; Work, Thierry M.; Balazs, George H.; Sutton, Claudia A.; Casey, Rufina N.; Casey, James W.

2004-01-01

Fibropapillomatosis (FP) of marine turtles is a neoplastic disease of ecological concern. A fibropapilloma-associated turtle herpesvirus (FPTHV) is consistently present, usually at loads exceeding one virus copy per tumor cell. DNA from an array of parasites of green turtles (Chelonia mydas) was examined with quantitative PCR (qPCR) to determine whether any carried viral loads are sufficient to implicate them as vectors for FPTHV. Marine leeches (Ozobranchus spp.) were found to carry high viral DNA loads; some samples approached 10 million copies per leech. Isopycnic sucrose density gradient/qPCR analysis confirmed that some of these copies were associated with particles of the density of enveloped viruses. The data implicate the marine leech Ozobranchus as a mechanical vector for FPTHV. Quantitative RT-PCR analysis of FPTHV gene expression indicated that most of the FPTHV copies in a fibropapilloma have restricted DNA polymerase expression, suggestive of latent infection

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)

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

An ion quencher operated lamp for multiplexed fluorescent bioassays.

Science.gov (United States)

Qing, Taiping; Sun, Huanhuan; He, Xiaoxiao; Huang, Xiaoqin; He, Dinggeng; Bu, Hongchang; Qiao, Zhenzhen; Wang, Kemin

2018-02-01

A novel and adjustable lamp based on competitive interaction among dsDNA-SYBR Green I (SGI), ion quencher, and analyte was designed for bioanalysis. The "filament" and switch of the lamp could be customized by employing different dsDNA and ion quencher. The poly(AT/TA) dsDNA was successfully screened as the most effective filament of the lamp. Two common ions, Hg 2+ and Fe 3+ , were selected as the model switch, and the corresponding ligand molecules cysteine (Cys) and pyrophosphate ions (PPi) were selected as the targets. When the fluorescence-quenched dsDNA/SGI-ion complex was introduced into a target-containing system, ions could be bound by competitive molecules and separate from the complex, thereby lighting the lamp. However, no light was observed if the biomolecule could not snatch the metal ions from the complex. Under the optimal conditions, sensitive and selective detection of Cys and PPi was achieved by the lamp, with practical applications in fetal bovine serum and human urine. This ion quencher regulated lamp for fluorescent bioassays is simple in design, fast in operation, and is more convenient than other methods. Significantly, as many molecules could form stable complexes with metal ions selectively, this ion quencher operated lamp has potential for the detection of a wide spectrum of analytes. Graphical abstract A novel and adjustable lamp on the basis of competitive interaction among dsDNA-SYBR Green I, ions quencher and analyte was designed for bioanalysis. The filament and switch of lamp could be customized by employing different dsDNA and ions quencher.

Rapid detection of Echinococcus species by a high-resolution melting (HRM) approach.

Science.gov (United States)

Santos, Guilherme Brzoskowski; Espínola, Sergio Martín; Ferreira, Henrique Bunselmeyer; Margis, Rogerio; Zaha, Arnaldo

2013-11-14

High-resolution melting (HRM) provides a low-cost, fast and sensitive scanning method that allows the detection of DNA sequence variations in a single step, which makes it appropriate for application in parasite identification and genotyping. The aim of this work was to implement an HRM-PCR assay targeting part of the mitochondrial cox1 gene to achieve an accurate and fast method for Echinococcus spp. differentiation. For melting analysis, a total of 107 samples from seven species were used in this study. The species analyzed included Echinococcus granulosus (n = 41) and Echinococcus ortleppi (n = 50) from bovine, Echinococcus vogeli (n = 2) from paca, Echinococcus oligarthra (n = 3) from agouti, Echinococcus multilocularis (n = 6) from monkey and Echinococcus canadensis (n = 2) and Taenia hydatigena (n = 3) from pig. DNA extraction was performed, and a 444-bp fragment of the cox1 gene was amplified. Two approaches were used, one based on HRM analysis, and a second using SYBR Green Tm-based. In the HRM analysis, a specific profile for each species was observed. Although some species exhibited almost the same melting temperature (Tm) value, the HRM profiles could be clearly discriminated. The SYBR Green Tm-based analysis showed differences between E. granulosus and E. ortleppi and between E. vogeli and E. oligarthra. In this work, we report the implementation of HRM analysis to differentiate species of the genus Echinococcus using part of the mitochondrial gene cox1. This method may be also potentially applied to identify other species belonging to the Taeniidae family.

Simultaneous fitting of real-time PCR data with efficiency of amplification modeled as Gaussian function of target fluorescence

Directory of Open Access Journals (Sweden)

Lazar Andreas

2008-02-01

Full Text Available Abstract Background In real-time PCR, it is necessary to consider the efficiency of amplification (EA of amplicons in order to determine initial target levels properly. EAs can be deduced from standard curves, but these involve extra effort and cost and may yield invalid EAs. Alternatively, EA can be extracted from individual fluorescence curves. Unfortunately, this is not reliable enough. Results Here we introduce simultaneous non-linear fitting to determine – without standard curves – an optimal common EA for all samples of a group. In order to adjust EA as a function of target fluorescence, and still to describe fluorescence as a function of cycle number, we use an iterative algorithm that increases fluorescence cycle by cycle and thus simulates the PCR process. A Gauss peak function is used to model the decrease of EA with increasing amplicon accumulation. Our approach was validated experimentally with hydrolysis probe or SYBR green detection with dilution series of 5 different targets. It performed distinctly better in terms of accuracy than standard curve, DART-PCR, and LinRegPCR approaches. Based on reliable EAs, it was possible to detect that for some amplicons, extraordinary fluorescence (EA > 2.00 was generated with locked nucleic acid hydrolysis probes, but not with SYBR green. Conclusion In comparison to previously reported approaches that are based on the separate analysis of each curve and on modelling EA as a function of cycle number, our approach yields more accurate and precise estimates of relative initial target levels.

Green Campus Study by using 10 UNEP’s Green University Toolkit Criteria in IPB Dramaga Campus

Science.gov (United States)

Sisriany, Saraswati; Sitti Fatimah, Indung

2017-10-01

Campus landscape is an important part of campus life, because it is regarded as a physical manifestation of the value of a college. Green campus is a concept to build sustainable living practices that are environmentally friendly in educational institutions around the world, including in IPB Dramaga Campus. The main objective of this study is to identified and analyze IPB Dramaga Campus sustainability used green campus criteria from UNEP (United Nations Environment Programme). The methods stages are data collection, analysis and assessment, and recommendation as the synthesis. All the data analyzed with gap analysis, then it assess with Likert Scale scoring. The results showed that green level of IPB Dramaga Campus is classified as Moderate, with total score 32. The result from each criterias are, Energy, Carbon and Climate Change is Moderate; Water is Not Good; Waste is Moderate; Biodiversity and Ecosystem Services is Very Good; Planning Design & Development is Good; Procurement is Moderate; Green Office is Very Not Good; Green Lab is Moderate; Green IT is Good; and Transport is Good. The Green Level of IPB Dramaga Campus will reach Very Good if these recommendation of strategies applied. The strategies are Green Office, Green Campus Audit, Green Champion, Green Financial Strategies, Water Treatment, Green Lab dan Off Campus Transportation.

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...

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.)

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.

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.

Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction.

Science.gov (United States)

Petersen, M I; Alvarez, I; Trono, K G; Jaworski, J P

2018-04-11

The detection of bovine leukemia virus (BLV) proviral DNA is an important tool to address whether an animal is infected with BLV. Compared with serological assays, real-time PCR accounts for greater sensitivity and can serve as a confirmatory test for the clarification of inconclusive or discordant serological test results. However, the high cost related to real-time PCR assays has limited their systematic inclusion in BLV surveillance and eradication programs. The aim of the present study was to validate a low-cost quantitative real-time PCR. Interestingly, by using SYBR Green detection dye, we were able to reduce the cost of a single reaction by a factor of 5 compared with most common assays based on the use of fluorogenic probes (i.e., TaqMan technology). This approach allowed a highly sensitive and specific detection and quantification of BLV proviral DNA from purified peripheral blood leukocytes and a milk matrix. Due to its simplicity and low cost, our in-house BLV SYBR quantitative real-time PCR might be used either as a screening or as a confirmatory test in BLV control programs. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

Are green cities healthy and equitable? Unpacking the relationship between health, green space and gentrification.

Science.gov (United States)

Cole, Helen V S; Garcia Lamarca, Melisa; Connolly, James J T; Anguelovski, Isabelle

2017-11-01

While access and exposure to green spaces has been shown to be beneficial for the health of urban residents, interventions focused on augmenting such access may also catalyse gentrification processes, also known as green gentrification. Drawing from the fields of public health, urban planning and environmental justice, we argue that public health and epidemiology researchers should rely on a more dynamic model of community that accounts for the potential unintended social consequences of upstream health interventions. In our example of green gentrification, the health benefits of greening can only be fully understood relative to the social and political environments in which inequities persist. We point to two key questions regarding the health benefits of newly added green space: Who benefits in the short and long term from greening interventions in lower income or minority neighbourhoods undergoing processes of revitalisation? And, can green cities be both healthy and just? We propose the Green Gentrification and Health Equity model which provides a framework for understanding and testing whether gentrification associated with green space may modify the effect of exposure to green space on health. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Green Energy for Green Economy: The Case Study of Kurdistan Region of Iraq (KRI

Directory of Open Access Journals (Sweden)

Ahmet Sekreter

2017-12-01

Full Text Available Green economy is an overarching purpose for sustainable growth and friendly environment. Renewable energy focuses on clean energy and sustainable development targets a continuous growth. Green economy includes both of them. Kurdistan Region of Iraq (KRI struggles serious problems in terms of economics after showing a remarkable economic growth until mid-2014. The increasing gap between demand and supply is seen another serious problem for KRI. Green energy is one of the essential stage towards to the green economy and it is one of the vital issue to succeed on the way of green economy. Solar energy is one of the fastest growing renewable energy source around the world and KRI has a great potential for solar energy. This study aims to stimulate KRI to invest green energy and encourage it to establish green economy to make its economy robust for the shocks and enable to show a sustainable development.

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.

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

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

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

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 ...

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...

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

Transcriptional profiles of chicken embryo cell cultures following infection with infectious bursal disease virus

DEFF Research Database (Denmark)

Li, Yiping; Handberg, K.J.; Juul-Madsen, H.R.

2007-01-01

Infectious bursal disease virus (IBDV) is the causative agent of infectious bursal disease in chickens and causes a significant economic loss for the poultry industry. Little is understood about the mechanism involved in the host responses to IBDV infection. For better understanding the IBDV......-host interaction, we measured steady-state levels of transcripts from 28 cellular genes of chicken embryo (CE) cell cultures infected with IBDV vaccine stain Bursine-2 during a 7-day infection course by use of the quantitative real-time RT-PCR SYBR green method. Of the genes tested, 21 genes (IRF-1, IFN 1...

Progesterone receptor isoform analysis by quantitative real-time polymerase chain reaction in formalin-fixed, paraffin-embedded canine mammary dysplasias and tumors

DEFF Research Database (Denmark)

Guil-Luna, S.; Stenvang, Jan; Brünner, Nils

2014-01-01

and its isoforms in formalin-fixed, paraffin-embedded tissue samples from canine mammary lesions (4 dysplasias, 10 benign tumors, and 46 carcinomas) using 1-step SYBR Green quantitative real-time polymerase chain reaction (RT-qPCR). Progesterone receptor was expressed in 75% of dysplasias, all benign...... in the expression of isoform A versus B. Analysis of progesterone receptor mRNA isoforms by RT-qPCR was successful in routinely formalin-fixed, paraffin-embedded tissue samples and enabled the distribution of isoforms A and B to be identified for the first time in dysplasias, benign tumors, and malignant tumors...

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.

Decay Of Bacterial Pathogens, Fecal Indicators, And Real-Time Quantitative PCR Genetic Markers In Manure-Amended Soils

Science.gov (United States)

This study examined persistence and decay of bacterial pathogens, fecal indicator bacteria (FIB), and emerging real-time quantitative PCR (qPCR) genetic markers for rapid detection of fecal pollution in manure-amended agricultural soils. Known concentrations of transformed green...

Decay Of Bacterial Pathogen, Fecal Indicators, And Real-Time Quantitative PCR Genetic Markers In Manure Amended Soils

Science.gov (United States)

This study examined persistence and decay of bacterial pathogens, fecal indicator bacteria, and emerging real-time quantitative PCR (qPCR) genetic markers for rapid detection of fecal pollution in manre-amended agricultural soils. Known concentrations of transformed green fluore...

Green Economy Performance and Green Productivity Growth in China’s Cities: Measures and Policy Implication

Directory of Open Access Journals (Sweden)

Jianglong Li

2016-09-01

Full Text Available Resource depletion and environmental degradation have become serious challenges for China’s sustainable development. This paper constructs indicators to assess China’s green economy performance and green productivity growth, in which economic expansion, resource conservation and environmental protection need to be incorporated simultaneously. For this purpose, we combine non-radial directional distance function and meta-frontier Malmquist productivity to develop the indicators. The methodology also allows for the decomposition of driving forces of China’s green economy. Moreover, the dataset employed in this paper allows for the evaluation of 275 cities in China during the period 2003–2012. The main findings are as follows. First, most of China’s cities did not perform efficiently in terms of the green economy, with an average score of only 0.233. Second, the growth rate of green productivity is slower than real GDP, and the green productivity growth in China is only moderate. Third, innovation is the main driving force of China’s green productivity growth, but the central region lags behind when it comes to green innovation. Fourth, artificial local protectionism and transport limitations impede the progress of cities that perform ineffectively in the green economy. Based on our empirical findings, we provide policy implications and suggestions for enhancing China’s green economy performance and productivity growth.

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.

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

c-DNA of HIV-1 detection on spot of Buffy-Coat of leukocytes (DBCS

Directory of Open Access Journals (Sweden)

Marco Rossi de Gasperis

2010-03-01

Full Text Available Introduction:The elective way for the diagnosis of HIV-1-infection in the window period and in children under the age of 16-18 months is to search virus integrated in leukocytes. Aim of the study was to assess the sensitivity and specificity of extraction from Buffy-Dried Coat Spot (DBCS in leukocyte to detect c-DNA with nested-PCR in HIV-1-infected individuals compared to Dried Blood Spot (DBS both extracted by automated instrument EZ1 (QIAGEN, Hilden, Germany. Both DBCS and both DBS were compared with those tests from whole blood by conventional DNA-extraction Methods: Five ml of whole blood from 50 HIV-infected individuals were collected. 40 μl of each sample were spotted on “FTA ELUTE Micro Card” (Whatman, Inc., Clifton, NJ, 200 μl were extracted according to the manual procedure (QIAGEN “QIAamp DNA minikit and the remaining sample was incubated at 37 °C for 120 minutes. Plasma was centrifuged at 1000 rcf/1g for 10 minutes at room temperature. Forty μl of the obtained buffy-coat was spotted. Both DBCS and both DBS were dried at room temperature for 24 hours.Two of 5 punch from each spot were extracted with TISSUE DNA kit (Biorobot EZ1 DSP “Qiagen” and eluted in 50 μl of buffer.The recovery of genomic DNA was measured amplifying the ß-globin gene by Real-Time “SybrGreen I”.The DNA was amplified for the “pol” gene of HIV-1 by nested PCR and revealed in “SYBR-green I”. Eight HIV-antibody-negative samples were used as internal control. Results:The experimental protocol adopted for the DBCS showed high sensitivity and specificity.The extracted DNA from DBS and DBCS was characterized by excellent quality and without any inhibitory agents. The amount of proviral DNA extracted from DBCS is similar to that obtained by conventional extraction, while the DBS test was significantly less sensitive. Conclusion:These preliminary data suggest that the amount of c-DNA obtained with DBS technique is often not enough for the

The structure of mAG, a monomeric mutant of the green fluorescent protein Azami-Green, reveals the structural basis of its stable green emission

International Nuclear Information System (INIS)

Ebisawa, Tatsuki; Yamamura, Akihiro; Kameda, Yasuhiro; Hayakawa, Kou; Nagata, Koji; Tanokura, Masaru

2010-01-01

The crystal structure of a monomeric mutant of Azami-Green (mAG) from G. fascicularis was determined at 2.2 Å resolution. Monomeric Azami-Green (mAG) from the stony coral Galaxea fascicularis is the first known monomeric green-emitting fluorescent protein that is not a variant of Aequorea victoria green fluorescent protein (avGFP). These two green fluorescent proteins are only 27% identical in their amino-acid sequences. mAG is more similar in its amino-acid sequence to four fluorescent proteins: Dendra2 (a green-to-red irreversibly photoconverting fluorescent protein), Dronpa (a bright-and-dark reversibly photoswitchable fluorescent protein), KikG (a tetrameric green-emitting fluorescent protein) and Kaede (another green-to-red irreversibly photoconverting fluorescent protein). To reveal the structural basis of stable green emission by mAG, the 2.2 Å crystal structure of mAG has been determined and compared with the crystal structures of avGFP, Dronpa, Dendra2, Kaede and KikG. The structural comparison revealed that the chromophore formed by Gln62-Tyr63-Gly64 (QYG) and the fixing of the conformation of the imidazole ring of His193 by hydrogen bonds and van der Waals contacts involving His193, Arg66 and Thr69 are likely to be required for the stable green emission of mAG. The crystal structure of mAG will contribute to the design and development of new monomeric fluorescent proteins with faster maturation, brighter fluorescence, improved photostability, new colours and other preferable properties as alternatives to avGFP and its variants

To Green or Not to Green? Evaluation of Green Stormwater Infrastructure in Kansas City Middle Blue River Project

Science.gov (United States)

The City of Kansas City, Mo., Water Services Department is implementing a pilot project to measure and evaluate the performance of green infrastructure. Information obtained through this pilot project will be used to guide the design of green solutions throughout Kansas City und...

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…

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...

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...

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...

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.

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

76 FR 35894 - Office of Federal High-Performance Green Buildings; Establishment of the Green Building Advisory...

Science.gov (United States)

2011-06-20

... Federal High-Performance Green Buildings; Establishment of the Green Building Advisory Committee AGENCY... announces the establishment of the Green Building Advisory Committee (the Committee), pursuant to Section... strategic plans, products and activities of the Office of Federal High-Performance Green Buildings and...

76 FR 65511 - Office of Governmentwide Policy; Office of Federal High-Performance Green Buildings; the Green...

Science.gov (United States)

2011-10-21

... Governmentwide Policy; Office of Federal High- Performance Green Buildings; the Green Building Advisory Committee... meeting of the Green Building Advisory Committee Meeting (the Committee). The meeting is open to the..., Office of Federal High-Performance Green Buildings, Office of Governmentwide Policy, General Services...

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...

Adult Learning Meets the Green Economy: Lessons from a Green Jobs Education Project

Science.gov (United States)

Wagner, Cecelia

2013-01-01

The new "green economy" affects adult education and workforce development as adult workers seek skills and knowledge that will help them find success in work and life. Recent years have brought about increased interest in and discussion of training for green jobs. Since the introduction of the Green Jobs Act in 2007, questions about how exactly to…

Planning the Green Walkable City: Conceptualizing Values and Conflicts for Urban Green Space Strategies in Stockholm

Directory of Open Access Journals (Sweden)

Hélène Littke

2015-08-01

Full Text Available Urban green spaces are essential elements of cities, contributing to the quality of life in numerous ways. However, densification strategies create a complex relationship between urban development and the quality, as well as the quantity, of urban green space. This paper examines the Green Walkable City Programme in Stockholm, a document developed to supplement the comprehensive plan as a strategic backbone for green urban planning. Based on interviews and content analysis, this paper identifies and discusses concerns raised in the development of the planning programme, and addresses the importance of urban green space for citizens’ well-being. The new comprehensive plan has introduced a shift in the attitude towards the urban green space in Stockholm. The need for urban growth is used to justify development of green fields, and a focus on the quality, rather than the quantity, of urban green space is promoted. Despite this progress, the public requests definitions for this quality approach and fears that nature within the city will be “parkified”. Therefore, this paper offers a critical reflection on the role of the Green Walkable City Programme, its situation within the context of Swedish green urban planning, and various areas of concern that have been highlighted.

Differential tissue expression of enhanced green fluorescent protein in ‘Green mice’

OpenAIRE

Ma, De-Fu; Tezuka, Hideo; Kondo, Tetsuo; Sudo, Katsuko; Niu, Dong-Feng; Nakazawa, Tadao; Kawasaki, Tomonori; Yamane, Tetsu; Nakamura, Nobuki; Katoh, Ryohei

2010-01-01

In order to clarify tissue expression of enhanced green fluorescent protein (EGFP) in ‘green mice’ from a transgenic line having an EGFP cDNA under the control of a chicken beta-actin promoter and cytomegalovirus enhancer, we studied the expression of EGFP in various organs and tissues from these ‘green mice’ by immunohistochemistry with anti- EGFP antibody in conjunction with direct observation for EGFP fluorescence using confocal laser scanning microscopy. On i...

Quick Green Scan: A Methodology for Improving Green Performance in Terms of Manufacturing Processes

Directory of Open Access Journals (Sweden)

Aldona Kluczek

2017-01-01

Full Text Available The heating sector has begun implementing technologies and practices to tackle the environmental and social–economic problems caused by their production process. The purpose of this paper is to develop a methodology, “the Quick-Green-Scan”, that caters for the need of quick assessment decision-makers to improve green manufacturing performance in companies that produce heating devices. The study uses a structured approach that integrates Life Cycle Assessment-based indicators, framework and linguistic scales (fuzzy numbers to evaluate the extent of greening of the enterprise. The evaluation criteria and indicators are closely related to the current state of technology, which can be improved. The proposed methodology has been created to answer the question whether a company acts on the opportunity to be green and whether these actions are contributing towards greening, maintaining the status quo or moving away from a green outcome. Results show that applying the proposed improvements in processes helps move the facility towards being a green enterprise. Moreover, the methodology, being particularly quick and simple, is a practical tool for benchmarking, not only in the heating industry, but also proves useful in providing comparisons for facility performance in other manufacturing sectors.

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.

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...

Building green supply chains in eco-industrial parks towards a green economy: Barriers and strategies.

Science.gov (United States)

Li, Jacqueline; Pan, Shu-Yuan; Kim, Hyunook; Linn, Jean H; Chiang, Pen-Chi

2015-10-01

As suggested by UNEP, the key to sustainable development is to create a "green economy" which should encapsulate all three sectors: the industry, the people, and the government. Therefore, there is an urgent need to develop and implement the green technologies into the existing facilities, especially in the developing countries. In this study, the role of green supply chains in eco-industrial parks (EIPs) towards a green economy was investigated. The strategies and effective evaluation procedures of the green economy were proposed by assessing the barriers from the perspective of institution, regulation, technology, and finance. In addition, three case studies from iron and steel-making, paper mill and pulping, and petrochemical industries were presented and illustrated for building the green supply chains. For example, in the case of Lin-Hai Industrial Park, a total of 15 efficient green supply chains using waste-to-resources technologies were established by 2012, resulting in an economic benefit of USD 100 million per year. It suggests that the green supply chains should be established to achieve both economic growth and environmental protection. With these successful experiences, building a green supply chain within industrial park should be extensively promoted to make traditional industries around the world being environmentally bearable, economic viable, and social equitable. Copyright © 2015 Elsevier Ltd. All rights reserved.

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

Effects of Technological Innovation in Relationship between Green Supply Chain Management Practices and Green Performance

OpenAIRE

Umar, Mohammed Sangiru; Danjuma, Ibrahim; Hammawa, Dahiru Dauda; Habibu, Sherif Ahmed

2016-01-01

Although scholars have conceptualised on green supply chain management practices and green performance, evidence to validate the conceptualisation was lacking, albeit in the context of small and medium enterprises. In addition, effect of technological innovation on supply chain practices and green performance was largely unexplored by researchers. Therefore this study validates and provides empirical evidence on the relationship between green supply chain management practices, technological i...

Multi-scale monitoring of a remarkable green roof: the Green Wave of Champs-sur-Marne

Science.gov (United States)

Stanic, Filip; Versini, Pierre-Antoine; Schertzer, Daniel; Delage, Pierre; Tchiguirinskaia, Ioulia; Cui, Yu-Jun; Baudoin, Genevieve

2017-04-01

The installation of green infrastructures on existing or new roofs has become very popular in recent years (more than 2 km2 of green roofs is implemented each year in France) for many reasons. Among all of the green roofs' advantages, those related to storm water management are often pushed forward, since it has been pointed out that urban runoff peak can be significantly reduced and delayed thanks to the green roofs' retention and detention capabilities. Microclimate can also be affected by decreasing the temperature in the surrounding green area. However, dynamic physical processes involved in green roofs are highly non linear and variable. In order to accurately assess their performances, detailed monitoring experiments are required, both in situ and in the lab, so as to better understand the thermo-hydric behaviour of green roofs and to capture the related spatio-temporal variability at different scales. Based on these considerations, the 1 ha area wavy-form green roof of a section of the Bienvenüe building, called the Green Wave, is currently being monitored in Champs-sur-Marne (France), in front of Ecole des Ponts ParisTech. Initiated in the "Blue Green Dream" European project, detailed measurements systems have been implemented for studying all components of the water balance. Among others, a wireless network of water content and temperature sensors has been especially installed for characterizing spatial and temporal variability of infiltration, retention and evapotranspiration processes. In parallel, some laboratory tests have been conducted to better characterize the hydro-mechanical properties of the substrate. Moreover, at the Green Wave scale, some discharge measurements are carried out in the storm-water pipes that are collecting drained water, to determine runoff flow. This talk will present the current monitoring campaigns and analyze the data collected in the Universal Multifractal framework. This work represents the initial stage for developing a

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

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...

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.

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

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

Green Skills for Green Economy: Case of the Environmental Education Role in Kazakhstan's Economy

Science.gov (United States)

Dlimbetova, Gaini; Zhylbaev, Zhanbol; Syrymbetova, Lyailya; ?liyeva, Aiman

2016-01-01

The research on situation with developing "green skills" in conditions of transition to "green economy" is analysed in this article. Kazakhstan like many other states has been going through transition to "green economy" since 2013. Economic reforms have made an impact on the system of environmental education. The…

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.

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...

A review of drug delivery systems based on nanotechnology and green chemistry: green nanomedicine.

Science.gov (United States)

Jahangirian, Hossein; Lemraski, Ensieh Ghasemian; Webster, Thomas J; Rafiee-Moghaddam, Roshanak; Abdollahi, Yadollah

2017-01-01

This review discusses the impact of green and environmentally safe chemistry on the field of nanotechnology-driven drug delivery in a new field termed "green nanomedicine". Studies have shown that among many examples of green nanotechnology-driven drug delivery systems, those receiving the greatest amount of attention include nanometal particles, polymers, and biological materials. Furthermore, green nanodrug delivery systems based on environmentally safe chemical reactions or using natural biomaterials (such as plant extracts and microorganisms) are now producing innovative materials revolutionizing the field. In this review, the use of green chemistry design, synthesis, and application principles and eco-friendly synthesis techniques with low side effects are discussed. The review ends with a description of key future efforts that must ensue for this field to continue to grow.

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...

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.

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.

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

EnviroAtlas - Green Bay, WI - Estimated Percent Green Space Along Walkable Roads

Data.gov (United States)

U.S. Environmental Protection Agency — This EnviroAtlas dataset estimates green space along walkable roads. Green space within 25 meters of the road centerline is included and the percentage is based on...

An Empirical Study on Green Innovation Efficiency in the Green Institutional Environment

Directory of Open Access Journals (Sweden)

Yang Gao

2018-03-01

Full Text Available Previous studies have found that reverse technology spillover effects can promote industrial technology modernization in developing countries. However, it is still unknown whether reverse technology spillover effects can improve green innovation efficiency in developing countries. In particular, institutional uncertainties characteristic of transition economies have a significant impact on industrial modernization. Therefore, researching the impact of the institutional environment on the relationship between reverse technology spillover effects and green innovation efficiency is of great significance. In this paper, we use data from G20 countries as well as China’s foreign direct investment (FDI data to measure the effects of reverse technology spillovers and adopt the threshold effect model to explore the relationship between reverse technology spillover effects and green innovation efficiency as well as the influence of the institutional environment on this relationship, based on China’s provincial panel data from 2003 to 2015. The empirical results show that the reverse technology spillover effects can effectively improve green innovation efficiency. There is a threshold for the influence of the institutional environment on the relationship between reverse technology spillover effects and green innovation efficiency. When the institutional development level surpasses the threshold value, an acceleration effect is generated. In addition, we find that the legal system is the key bottleneck in terms of improving green innovation efficiency. How to improve and perfect the path of institutional construction in China and how to enable institutions to gain threshold speed-up effects have become the major problems the Chinese government faces in institutional construction. The research results of this paper offer a reference to developing countries in regard to improving their institutions and enhancing their green innovation efficiency.

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.

Green gold. 15 tax proposals for a green and innovative economy

International Nuclear Information System (INIS)

Van Engelen, D.; Wit, R.; Blaauw, K.; Winckers, J.

2010-06-01

This publication contains 15 proposals for green taxes in the Dutch economy. The benefit of these 15 proposals is over 11 billion euros per year and leads to a reduction of CO2 emissions of at least 12.5 megatons per year. Greening taxes involves a budget neutral shift from taxing labor and profits to taxing environmental pollution and the depletion of natural resources. The proposals reward businesses and citizens which invest in the development and application of innovative green solutions. This leads to an improvement of climate, environment and nature as well as the competitiveness of the Dutch economy. [nl

A Real Time PCR Platform for the Simultaneous Quantification of Total and Extrachromosomal HIV DNA Forms in Blood of HIV-1 Infected Patients

Science.gov (United States)

Canovari, Benedetta; Scotti, Maddalena; Acetoso, Marcello; Valentini, Massimo; Petrelli, Enzo; Magnani, Mauro

2014-01-01

Background The quantitative measurement of various HIV-1 DNA forms including total, unintegrated and integrated provirus play an increasingly important role in HIV-1 infection monitoring and treatment-related research. We report the development and validation of a SYBR Green real time PCR (TotUFsys platform) for the simultaneous quantification of total and extrachromosomal HIV-1 DNA forms in patients. This innovative technique makes it possible to obtain both measurements in a single PCR run starting from frozen blood employing the same primers and standard curve. Moreover, due to identical amplification efficiency, it allows indirect estimation of integrated level. To specifically detect 2-LTR a qPCR method was also developed. Methodology/Findings Primers used for total HIV-1 DNA quantification spanning a highly conserved region were selected and found to detect all HIV-1 clades of group M and the unintegrated forms of the same. A total of 195 samples from HIV-1 patients in a wide range of clinical conditions were analyzed with a 100% success rate, even in patients with suppressed plasma viremia, regardless of CD4+ or therapy. No significant correlation was observed between the two current prognostic markers, CD4+ and plasma viremia, while a moderate or high inverse correlation was found between CD4+ and total HIV DNA, with strong values for unintegrated HIV DNA. Conclusions/Significance Taken together, the results support the use of HIV DNA as another tool, in addition to traditional assays, which can be used to estimate the state of viral infection, the risk of disease progression and to monitor the effects of ART. The TotUFsys platform allowed us to obtain a final result, expressed as the total and unintegrated HIV DNA copy number per microgram of DNA or 104 CD4+, for 12 patients within two working days. PMID:25364909

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

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...

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.

Evaluation of green roof as green technology for urban stormwater quantity and quality controls

International Nuclear Information System (INIS)

Kok, K H; Sidek, L M; Basri, H; Muda, Z C; Beddu, S; Abidin, M R Z

2013-01-01

Promoting green design, construction, reconstruction and operation of buildings has never been more critical than now due to the ever increasing greenhouse gas emissions and rapid urbanizations that are fuelling climate change more quickly. Driven by environmental needs, Green Building Index (GBI) was founded in Malaysia to drive initiative to lead the property industry towards becoming more environment-friendly. Green roof system is one of the assessment criteria of this rating system which is under category of sustainable site planning and management. An extensive green roof was constructed in Humid Tropics Center (HTC) Kuala Lumpur as one of the components for Stormwater Management Ecohydrology (SME) in order to obtain scientific data of the system. This paper evaluates the performance of extensive green roof at Humid Tropics Center with respect to urban heat island mitigation and stormwater quantity and quality controls. Findings indicate that there was a reduction of around 1.5°C for indoor temperature of the building after installation of green roof. Simulations showed that the peak discharge was reduced up to 24% relative to impervious brown roof. The results show an increment of pH and high concentration of phosphate for the runoff generated from the green roof and the runoff water quality ranged between class I and II under INWQS.

Evaluation of green roof as green technology for urban stormwater quantity and quality controls

Science.gov (United States)

Kok, K. H.; Sidek, L. M.; Abidin, M. R. Z.; Basri, H.; Muda, Z. C.; Beddu, S.

2013-06-01

Promoting green design, construction, reconstruction and operation of buildings has never been more critical than now due to the ever increasing greenhouse gas emissions and rapid urbanizations that are fuelling climate change more quickly. Driven by environmental needs, Green Building Index (GBI) was founded in Malaysia to drive initiative to lead the property industry towards becoming more environment-friendly. Green roof system is one of the assessment criteria of this rating system which is under category of sustainable site planning and management. An extensive green roof was constructed in Humid Tropics Center (HTC) Kuala Lumpur as one of the components for Stormwater Management Ecohydrology (SME) in order to obtain scientific data of the system. This paper evaluates the performance of extensive green roof at Humid Tropics Center with respect to urban heat island mitigation and stormwater quantity and quality controls. Findings indicate that there was a reduction of around 1.5°C for indoor temperature of the building after installation of green roof. Simulations showed that the peak discharge was reduced up to 24% relative to impervious brown roof. The results show an increment of pH and high concentration of phosphate for the runoff generated from the green roof and the runoff water quality ranged between class I and II under INWQS.

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...

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.

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.

Discussion Tourism Industry on Energy of Green Tourism and Green Hotel

Directory of Open Access Journals (Sweden)

Wang Zeyung

2016-01-01

Full Text Available Tourism industry is closely linked with the natural environment but with a highly indivisibility of symbiotic relationship. Green tourism and green tourism hotel are not only the spindle stage of development industry. The environmental protection is also an environmental conservation and sustainable development of substantive liability demonstration. The study is also belong to the substance RDF itself, so we can call “clean energy”. The raw materials came from agricultural waste through proper blending ratio and control technology, after PP14 adhesive extruded through the fluidized bed pyrolysis cracking process to burn stability. The recovery can also be used as fuel volatile process of drying and gasification. However, in the actual economic cost of the test running the hotel industry can reduce the cost per MJ USD $ 0.0082, more economical than coal expenses 23.17% of the fuel. Therefore, green hotel through biomass fuels RDF as clean fuels can further reduce carbon emissions to reach the green hotel of expectations.

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.

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…

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

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

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...

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.

Game Analysis and Countermeasures Discussion on Green Marketing

Institute of Scientific and Technical Information of China (English)

2011-01-01

On the basis of making certain assumption on the game situation of carrying out green marketing, this paper conducts game analysis on the green marketing choice among enterprises, the green marketing choice between enterprises and consumers, and the green marketing choice of consumers. Then this paper expounds the necessity of implementing green marketing as follows: the green marketing is the inevitable requirements of sustainable development of economy; the green marketing is the inevitable choice of green consumption mode; the green marketing is the inevitable results of legalization of environmental problems. The problems faced by the implementation of green marketing are analyzed as follows: first, the concept of green marketing has not yet been established; second, the sociality of green demand has not yet taken shape; third, production characteristic of green products has not yet formed. The countermeasures of implementing green marketing as follows: pay attention to the propaganda and education of modern marketing concept; regulate the competition in the market of green products; strengthen transparency of green market information; reinforce the legislation work of food safety.

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.

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...

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.

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.

Effect of Low Temperature and Wheat Winter-Hardiness on Survival of Puccinia striiformis f. sp. tritici under Controlled Conditions.

Directory of Open Access Journals (Sweden)

Lijie Ma

Full Text Available Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst, is one of the most important diseases of wheat worldwide. Understanding the survival of Pst during the overwintering period is critical for predicting Pst epidemics in the spring. Real-time quantitative PCR (qPCR methods quantifying Pst DNA and RNA (cDNA were developed and compared for the ability to quantify viable Pst in leaf tissues. Both qPCR of DNA and RNA can provide reliable measurement of viable Pst in plant tissues prior to the late sporulation stage for which qPCR of DNA gave a much higher estimate of fungal biomass than qPCR of RNA. The percentage of Pst biomass that was viable in detached and attached leaves under low temperatures decreased over time. Pst survived longer on attached leaves than on detached leaves. The survival of Pst in cultivars with strong winter-hardiness at 0°C and -5°C was greater than those with weak winter-hardiness. However, such differences in Pst survival among cultivars were negligible at -10, -15 and -20°C. Results indicated that Pst mycelia inside green leaves can also be killed by low temperatures rather than through death of green leaves under low temperatures. The relationship of Pst survival in attached leaves with temperature and winter-hardiness was well described by logistic models. Further field evaluation is necessary to assess whether inclusion of other factors such as moisture and snow cover could improve the model performance in predicting Pst overwintering potential, and hence the epidemic in spring.

No greens in the forest? Note on the limited consumption of greens in the Amazon

Directory of Open Access Journals (Sweden)

Esther Katz

2012-12-01

Full Text Available The consumption of greens is reported as being very minor among Amazonian Indians. The authors of this article present a new review of this subject, based on fieldwork with Amerindians and other populations in different parts of the Brazilian Amazon and French Guiana. Written sources on Brazilian, Peruvian, Columbian and Venezuelan Amazon were also reviewed. The consumption of cultivated, semi-cultivated and wild species of greens was taken into account here, as the data specific to wild greens is very scarce. It is confirmed that greens are not commonly eaten among native Amazonians and that some ethnic groups do not consume them at all. The consumed species are usually young shoots of weeds or cassava leaves. Common in the Belém region are some specific aromatic plants, which have been diffused to other parts of the Amazon, together with introduced plants such as kale and coriander. Migrants from Northeastern Brazil settled in the Amazon consume some cultivated greens, especially aromatic plants. Maroons are the ones who use more greens in their diet. Native Amazonian people, who supplement agriculture with game and fish, follow a hunter-gatherer pattern, preferring wild fruit and tubers to greens.

Differential tissue expression of enhanced green fluorescent protein in 'green mice'.

Science.gov (United States)

Ma, De-Fu; Tezuka, Hideo; Kondo, Tetsuo; Sudo, Katsuko; Niu, Dong-Feng; Nakazawa, Tadao; Kawasaki, Tomonori; Yamane, Tetsu; Nakamura, Nobuki; Katoh, Ryohei

2010-06-01

In order to clarify tissue expression of enhanced green fluorescent protein (EGFP) in 'green mice' from a transgenic line having an EGFP cDNA under the control of a chicken beta-actin promoter and cytomegalovirus enhancer, we studied the expression of EGFP in various organs and tissues from these 'green mice' by immunohistochemistry with anti- EGFP antibody in conjunction with direct observation for EGFP fluorescence using confocal laser scanning microscopy. On immunohistochemical examination and on direct observation by confocal laser scanning microscopy, the level of EGFP expression varied among organs and tissues. EGFP expression was diffusely and strongly observed in the skin, pituitary, thyroid gland, parathyroid gland, heart, gall bladder, pancreas, adrenals and urinary bladder. There was only sporadic and weak expression of EGFP in the epithelium of the trachea, bronchus of the lung, stratified squamous epithelium and gastric glands of the stomach, hepatic bile ducts of the liver, glomeruli and renal tubules of the kidney and endo-metrial glands of the uterus. Furthermore, EGFP was only demonstrated within the goblet and paneth cells in the colon and small intestine, the tall columnar cells in the ductus epididymis, and the leydig cells in the testis. In conclusion, our results show that EGFP is differentially expressed in organs and tissues of 'green mice', which indicates that 'green mice' may prove useful for research involving transplantation and tissue clonality.

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…

Flow cytometric fingerprinting for microbial strain discrimination and physiological characterization.

Science.gov (United States)

Buysschaert, Benjamin; Kerckhof, Frederiek-Maarten; Vandamme, Peter; De Baets, Bernard; Boon, Nico

2018-02-01

The analysis of microbial populations is fundamental, not only for developing a deeper understanding of microbial communities but also for their engineering in biotechnological applications. Many methods have been developed to study their characteristics and over the last few decades, molecular analysis tools, such as DNA sequencing, have been used with considerable success to identify the composition of microbial populations. Recently, flow cytometric fingerprinting is emerging as a promising and powerful method to analyze bacterial populations. So far, these methods have primarily been used to observe shifts in the composition of microbial communities of natural samples. In this article, we apply a flow cytometric fingerprinting method to discriminate among 29 Lactobacillus strains. Our results indicate that it is possible to discriminate among 27 Lactobacillus strains by staining with SYBR green I and that the discriminatory power can be increased by combined SYBR green I and propidium iodide staining. Furthermore, we illustrate the impact of physiological changes on the fingerprinting method by demonstrating how flow cytometric fingerprinting is able to discriminate the different growth phases of a microbial culture. The sensitivity of the method is assessed by its ability to detect changes in the relative abundance of a mix of polystyrene beads down to 1.2%. When a mix of bacteria was used, the sensitivity was as between 1.2% and 5%. The presented data demonstrate that flow cytometric fingerprinting is a sensitive and reproducible technique with the potential to be applied as a method for the dereplication of bacterial isolates. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

Part-per-trillion level detection of estradiol by competitive fluorescence immunoassay using DNA/dye conjugate as antibody multiple labels.

Science.gov (United States)

Zhu, Shengchao; Zhang, Qin; Guo, Liang-Hong

2008-08-22

Fluorescent organic dyes are currently the standard signal-generating labels used in microarray quantification. However, new labeling strategies are needed to meet the demand for high sensitivity in the detection of low-abundance proteins and small molecules. In this report, a long-chain DNA/dye conjugate was used to attach multiple fluorescence labels on antibodies to improve signal intensity and immunoassay sensitivity. Compared with the 30 base-pair (bp) oligonucleotide used in our previous work [Q. Zhang, L.-H. Guo, Bioconjugate Chem. 18 (2007) 1668-1672], conjugation of a 219 bp DNA in solution with a fluorescent DNA binder SYBR Green I resulted in more than sixfold increase in signal intensity, consistent with the increase in bp number. In a direct immunoassay for the detection of goat anti-mouse IgG in a mouse IgG-coated 96-well plate, the long DNA conjugate label also produced higher fluorescence than the short one, accompanied by about 15-fold improvement in the detection limit. To demonstrate its advantage in real applications, the DNA/dye conjugate was employed in the competitive immunoassay of 17beta-estradiol, a clinically and environmentally important analyte. The biotin-terminated DNA was attached to biotinylated anti-estradiol antibody through the biotin/streptavidin/biotin bridge after the immuno-reaction was completed, followed by conjugation with SYBR Green I. The limit of detection for 17beta-estradiol is 1.9 pg mL(-1), which is 200-fold lower than the assay using fluorescein-labeled antibodies. The new multiple labeling strategy uses readily available reagents, and is also compatible with current biochip platform. It has great potential in the sensitive detection of protein and antibody microarrays.

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...

Opportunities for green growth; Vihreaen kasvun mahdollisuudet

Energy Technology Data Exchange (ETDEWEB)

Antikainen, R.; Mickwitz, P.; Seppaelae, J. [and others

2013-03-15

The report seeks an answer to the question as to the kind of policy decisions (steps) by which preconditions for green growth may be created in Finland. The proposed steps are based on a review of earlier research and studies relating to Finland's key consumption and production systems (food, housing, transport and energy) and to certain path finding countries in terms of the green economy (the Netherlands, Germany, Sweden, Brazil). In addition, the report examines various models by which systemic change towards a green economy may be supported. The report also highlights successful examples of green business activity and measures to promote green growth. Green Growth is defined as low-carbon, resource-efficient economic growth based on safeguarding the functional capacity of ecosystems while promoting wellbeing and social justice. Green growth is considered to have significant worldwide potential, which is currently evident particularly in the rapid growth of cleantech demand. Successful future actors will be more material- and energy-efficient than their competitors, and they will be able to provide services and products flexibly for a low-carbon society. There are opportunities for green growth in all sectors of society. Green growth may consist of an entirely new kind of business activity and create new companies, but there are also opportunities in our traditional energy- and resource-intensive industries. Companies have a key role in growth, but realising green growth also requires changes in consumption. Central, regional and local government will act as facilitators in creating the preconditions for green growth. The report presents a number of policy measures and processes by which Finland can support green growth. Proposals for steps towards green growth include: (A.) Creating preconditions for green growth through a joint vision and political commitment. (B.) Stimulating companies' green growth potential and boosting green demand. (C

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

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.

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.

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!

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.

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

Two shades of Green? The electorates of GreenLeft and the Party for the Animals

NARCIS (Netherlands)

Otjes, Simon; Krouwel, Andre

2015-01-01

The Netherlands has two electorally significant parties that might be considered to be part of the Green' family: GreenLeft and the Party for the Animals. These two parties appeal to different niches of the Green electorate, identified on the basis of issue dimensions, demographics, and their trust

Evaluation of carrier-mediated siRNA delivery

DEFF Research Database (Denmark)

Colombo, Stefano; Nielsen, Hanne Mørck; Foged, Camilla

2013-01-01

RNA delivery. An in vitro cell culture model system expressing enhanced green fluorescent protein (EGFP) was used to develop the assay, which was based on the intracellular quantification of a full-length double-stranded Dicer substrate siRNA by stem-loop RT qPCR. The result is a well-documented protocol......RNA delivered by use of carriers remains an analytical challenge. The purpose of the present study was to optimize and validate an analytical protocol based on stem-loop reverse transcription quantitative polymerase chain reaction (RT qPCR) to quantitatively monitor the carrier-mediated intracellular si...

An eye for the green top : an independent voice for green roofs in the UK

Energy Technology Data Exchange (ETDEWEB)

Frith, M.; Gedge, D. [Livingroofs.org, England (United Kingdom)

2005-07-01

Livingroofs.org is a non-profit organization that was established to provide a resource for promoting green roofs in the United Kingdom (UK). Current policies in the UK related to the planning and development of green roofs are a constraint to their uptake. The emerging emphasis on sustainable development is bringing about a revision of planning policies that may make green roofs more desirable. However, green roof design standards are still being developed by a largely unmonitored industry furthering a product with which most people are unfamiliar. This paper reviewed a number of issues that need to be addressed to assist in the wider adoption of green roofs, including increased awareness; an identification of the real benefits of green roofs; guidance and research; planning policies; fiscal incentives; industry standards and codes of practice. Details of the current policy frameworks for construction, urban design and climate change were also outlined. Two specific projects were reviewed to provide an insight into the way in which Livingroofs.org intends to promote green roof technology: a roof owned by the Birmingham city council and a collaboration with a Swiss partner to design a roof for the Komodo Dragon House at the London Zoo. It was concluded that there is now a real likelihood that the widespread adoption of green roofs will occur in the UK within the next 5 years, both in terms of new developments and the vast potential for retro-fitting on existing buildings. 47 refs., 2 tabs.

Safety assessment of green tea based beverages and dried green tea extracts as nutritional supplements.

Science.gov (United States)

Dekant, Wolfgang; Fujii, Kenkichi; Shibata, Eiichiro; Morita, Osamu; Shimotoyodome, Akira

2017-08-05

The safety of green tea infusions and green tea extract (GTE)-based products is reviewed regarding catechins. Epigallocatechin 3-gallate (EGCG), the major catechin present in green tea, is suspected of being responsible for liver toxicity reported in humans consuming food supplements. Intake of EGCG with green tea infusions and GTE-based beverages is up to about 450mg EGCG/person/day in Europe and higher in Asia. Consumption of green tea is not associated with liver damage in humans, and green tea infusion and GTE-based beverages are considered safe in the range of historical uses. In animal studies, EGCG's potency for liver effects is highly dependent on conditions of administration. Use of NOAELs from bolus administration to derive a tolerable upper intake level applying the margin of safety concept results in acceptable EGCG-doses lower than those from one cup of green tea. NOAELs from toxicity studies applying EGCG with diet/split of the daily dose are a better point of departure for risk characterization. In clinical intervention studies, liver effects were not observed after intakes below 600mg EGCG/person/day. Thus, a tolerable upper intake level of 300mg EGCG/person/day is proposed for food supplements; this gives a twofold safety margin to clinical studies that did not report liver effects and a margin of safety of 100 to the NOAELs in animal studies with dietary administration of green tea catechins. Copyright © 2017 Elsevier B.V. All rights reserved.

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)

Buying green! A handbook on green energy procurement. 2nd edition

Energy Technology Data Exchange (ETDEWEB)

NONE

2011-11-10

Over 2 trillion Euros is spent on public contracts on a yearly basis in the EU, translating to 19 percent of its GDP. ICLEI has been working with the European Commission to produce guidance on green public procurement (GPP) which will help public authorities to reduce their environmental impact, while complying with the procurement rules. The result is the new edition of the Buying Green! handbook. The handbook identifies strategies for GPP, and explains in detail how these can be implemented at each stage of the procurement process. It is intended for public sector procurers but is also a useful reference for policy makers and businesses who are either implementing their own green purchasing or responding to tenders. In addition to explaining how GPP can be implemented under the EU procurement rules, the handbook includes examples of green contracts from across the EU-27. The resource has been fully updated from the previous 2004 edition and includes expanded sections on life-cycle costing (LCC) and the construction, timber, electricity and food and drink sectors.

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

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.

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.

Analysis on the Relationship between Green Accounting and Green Design for Enterprises

Directory of Open Access Journals (Sweden)

Jui-Che Tu

2015-05-01

Full Text Available Green design is advocated and developed in response to the increasingly deteriorating global environment, but its implementation is only based on the morality of the entrepreneurs, without economic incentive and legal restraint. As a result, green design has not been widely adopted. In recent years, the European countries, the U.S., Japan, the UN and Taiwan have successively promoted environmental accounting guidelines and required enterprises to disclose environmental improvement information, so as to improve the environment through production that will unavoidably impact product manufacturing. How product design should respond to this trend is a concern of this study. This study adopted the KJ (Kawakita Jiro method and the meta-research method to analyze the influence factors. Then, it was discussed whether green design is feasible. The results showed that the requirements of green accounting include: expanding corporate social responsibility, production cannot be exempted from environmental protection, the manufacturing of clean products can generate pollution, the external production cost should be internalized, the redesign to improve the product production process and packaging, reducing resource waste and implementing the (Reduce, Recycle, Reuse 3R policy, lifecycle assessment for all assessments and developing environmentally-friendly products, which can be solved with green design.

GreenTalks at Boston Green Academy: Student Reflections on Performance Assessment

Science.gov (United States)

Kuriacose, Christina

2017-01-01

In spring 2017, for the third year running, 10th graders at Boston Green Academy (BGA) presented GreenTalks, a showcase of research on food justice issues. The day Christina Kuriacose visited the school, students were presenting the PowerPoints they had put together. All of them included a map plotting out the proximity of their neighborhood or…

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.

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...

Green tea phytocompounds as anticancer: A review

Directory of Open Access Journals (Sweden)

Najeeb Ullah

2016-04-01

Full Text Available Green tea is universally considered significant and its benefits have been experimentally explored by researchers and scientists. Anticancer potential of green tea has been completely recognized now. Green tea contains anti-cancerous constituents and nutrients that have powerful remedial effects. By using electronic data base (1998–2015, different compounds in green tea possessing anticancer activity including epigallocatechin-3-gallate, paclitaxel and docetaxel combinations, ascorbic acid, catechins, lysine, synergistic arginine, green tea extract, proline, and green tea polyphenols has been reported. Green tea extracts exhibited remedial potential against cancer of lung, colon, liver, stomach, leukemic cells, prostate, breast, human cervical cells, head, and neck. For centuries, green tea has been utilized as medicine for therapeutic purposes. It originated in China and extensively used in Asian countries for blood pressure depression and as anticancer medicine. Green tea has therapeutic potential against many diseases such as lowering of blood pressure, Parkinson’s disease, weight loss, esophageal disease, skin-care, cholesterol, Alzheimer’s disease and diabetes.

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.)

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.

Green Hotel Management and Green Star Practice: A Case Study of Best Western President Istanbul Hotel

OpenAIRE

ATAY, Lütfi; DİLEK, S. Emre; YILDIRIM, H. Mehmet

2014-01-01

Defined as environmentally-sensitive marketing, green marketing is applied as a green hotel by accommodation establishments, and as a result of this application, hotels are entitled to be awarded a green star certificate. While those international hotel establishments which have become a brand are carrying out important activities with respect to the green hotel practice, it might be stated that hotels in Turkey are at the beginning level concerning sensitivity to the environment. In the stud...

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...

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.

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.

Urban Green Network Design: Defining green network from an urban planning perspective

Directory of Open Access Journals (Sweden)

Andrea Tulisi

2017-08-01

Full Text Available From the theoretical context of Smart City various studies have emerged that adopt an analytical approach and description of urban phenomena based on the principles of “network design”; this line of research uses the network systems theory to define the principles that regulate the relationships among the various elements of urban sub-systems in order to optimize their functionality. From the same theoretical basis, urban greenspaces have also been studied as networks, by means of the creation of models capable of measuring the performance of the system in its entirety, posing the basis of a new multy-disciplinary research field called green network. This paper presents the results of research aimed at clarifying the meaning of green network from an urban planning perspective through a lexical analysis applied to a textual corpus of more than 300 abstracts of research papers that have dealt with this topic over the last twenty years. The results show that the concept of green network appears still fuzzy and unclear, due to the different meaning given to the term “green” and to an incorrect use of the term “network”, often referred to as a generic set of natural areas present in a city, without any reference to the network system theory or to the basic rules linking these elements together. For this reason, the paper proposes a unique definition of green network from an urban planning perspective that takes into account the contribution of other research areas to effective green infrastructure planning. This is the concept of “urban green network design” defined as “an urban planning practice, supported by decision support tools able to model green infrastructure as network, composed by natural and semi-natural areas, whose connections are modelled according to specific variables, in order to deliver an equal distribution of public services for enhancing the quality of life as well as a wide range of ecosystem services”.

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...

Determination of adulteration of malachite green in green pea and some prepared foodstuffs by micellar liquid chromatography.

Science.gov (United States)

Ashok, Vipin; Agrawal, Nitasha; Durgbanshi, Abhilasha; Esteve-Romero, Josep; Bose, Devasish

2014-01-01

A simple, fast, and robust micellar LC method was developed for the separation and identification of the nonpermitted color malachite green in green pea and some ready-to-eat foodstuffs. Malachite green (4-[(4-dimethylaminophenyl) phenyl-methyl]-N,N-dimethylaniline) is a hazardous dye that is used to treat fungal and protozoan infections in fish and is a common adulterant (coloring agent) in green pea and other green vegetables because of its green color. In the present work, malachite green was determined in various foodstuffs using a direct injection technique on an RP C18 column with isocratic elution. The optimum mobile phase consisted of 0.15 M sodium dodecyl sulfate (SDS), 6% pentanol buffered at pH 5. Detection was carried out at 620 nm. Malachite green was eluted in 9.2 min without any interference caused by endogenous compounds. Linearities (r > 0.9999), intraday and interday precision (RSD less than 1.00%) in micellar media, and robustness were studied for method validation. LOD and LOQ were 0.10 and 0.25 ppm, respectively. The simplicity of the developed method makes it useful for routine analysis in the area of food QC.

Optical Determination of Lead Chrome Green in Green Tea by Fourier Transform Infrared (FT-IR Transmission Spectroscopy.

Directory of Open Access Journals (Sweden)

Xiaoli Li

Full Text Available The potential of Fourier transform infrared (FT-IR transmission spectroscopy for determination of lead chrome green in green tea was investigated based on chemometric methods. Firstly, the qualitative analysis of lead chrome green in tea was performed based on partial least squares discriminant analysis (PLS-DA, and the correct rate of classification was 100%. And then, a hybrid method of interval partial least squares (iPLS regression and successive projections algorithm (SPA was proposed to select characteristic wavenumbers for the quantitative analysis of lead chrome green in green tea, and 19 wavenumbers were obtained finally. Among these wavenumbers, 1384 (C = C, 1456, 1438, 1419(C = N, and 1506 (CNH cm-1 were the characteristic wavenumbers of lead chrome green. Then, these 19 wavenumbers were used to build determination models. The best model was achieved by least squares support vector machine (LS-SVMalgorithm with high coefficient of determination and low root-mean square error of prediction set (R2p = 0.864 and RMSEP = 0.291. All these results indicated the feasibility of IR spectra for detecting lead chrome green in green tea.

Green space definition affects associations of green space with overweight and physical activity

NARCIS (Netherlands)

Klompmaker, Jochem O.; Hoek, Gerard; Bloemsma, Lizan D.; Gehring, Ulrike; Strak, Maciej; Wijga, Alet H.; van den Brink, Carolien; Brunekreef, Bert; Lebret, Erik; Janssen, Nicole A.H.

Introduction In epidemiological studies, exposure to green space is inconsistently associated with being overweight and physical activity, possibly because studies differ widely in their definition of green space exposure, inclusion of important confounders, study population and data

A review of drug delivery systems based on nanotechnology and green chemistry: green nanomedicine

Directory of Open Access Journals (Sweden)

Jahangirian H

2017-04-01

Full Text Available Hossein Jahangirian,1 Ensieh Ghasemian Lemraski,2 Thomas J Webster,1 Roshanak Rafiee-Moghaddam,3 Yadollah Abdollahi4 1Department of Chemical Engineering, Northeastern University, Boston, MA, USA; 2Department of Chemistry, Faculty of Science, Ilam University, Ilam, Iran; 3School of Chemical Sciences and Food Technology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Selangor, 4Department of Electrical Engineering, Faculty of Engineering, University of Malaysia, Kuala Lumpur, Malaysia Abstract: This review discusses the impact of green and environmentally safe chemistry on the field of nanotechnology-driven drug delivery in a new field termed “green nanomedicine”. Studies have shown that among many examples of green nanotechnology-driven drug delivery systems, those receiving the greatest amount of attention include nanometal particles, polymers, and biological materials. Furthermore, green nanodrug delivery systems based on environmentally safe chemical reactions or using natural biomaterials (such as plant extracts and microorganisms are now producing innovative materials revolutionizing the field. In this review, the use of green chemistry design, synthesis, and application principles and eco-friendly synthesis techniques with low side effects are discussed. The review ends with a description of key future efforts that must ensue for this field to continue to grow. Keywords: green chemistry, cancer, drug delivery, nanoparticle

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.

GREEN GALAXIES IN THE COSMOS FIELD

International Nuclear Information System (INIS)

Pan, Zhizheng; Kong, Xu; Fan, Lulu

2013-01-01

We present research on the morphologies, spectra, and environments of ≈2350 'green valley' galaxies at 0.2 + color is used to define 'green valley'; it removes dusty star-forming galaxies from galaxies that are truly transitioning between the blue cloud and the red sequence. Morphological parameters of green galaxies are intermediate between those of blue and red galaxy populations, both on the Gini-asymmetry and the Gini-M 20 planes. Approximately 60%-70% of green disk galaxies have intermediate or big bulges, and only 5%-10% are pure disk systems, based on morphological classification using the Zurich Estimator of Structural Types. The obtained average spectra of green galaxies are intermediate between blue and red ones in terms of [O II], Hα, and Hβ emission lines. Stellar population synthesis on the average spectra shows that green galaxies are on average older than blue galaxies but younger than red galaxies. Green galaxies and blue galaxies have similar projected galaxy density (Σ 10 ) distributions at z > 0.7. At z * 10.0 M ☉ green galaxies located in a dense environment are found to be significantly larger than those of blue galaxies. The morphological and spectral properties of green galaxies are consistent with the transitioning population between the blue cloud and the red sequence. The possible mechanisms for quenching star formation activities in green galaxies are discussed. The importance of active galactic nucleus feedback cannot be well constrained in our study. Finally, our findings suggest that environmental conditions, most likely starvation and harassment, significantly affect the transformation of M * 10.0 M ☉ blue galaxies into red galaxies, especially at z < 0.5

Green paper with green electricity? Greening strategies of Nordic pulp and paper industry

International Nuclear Information System (INIS)

Luukkanen, Jyrki

2003-01-01

The article studies the opinions of paper producers in Finland and Norway and Finnish power producers about the eco-labelling of electricity and its possible effects on pulp and paper industry. The point of departure in the study is how large industrial consumers mediate concerns of environmental issues to the producers. Based on interviews of environmental, energy/power and marketing sector representatives of the companies the article analyses different views related to the criteria of green labelling, green electricity and papermaking, energy sources as image sources, environmental image of papermaking and the threats and opportunities the companies face in the changing international context. The analysis of the interviews is contextualised in the endogenous market based regulation framework of electricity market regulation

The Influence of Green Viral Communications on Green Purchase Intentions: The Mediating Role of Consumers’ Susceptibility to Interpersonal Influences

Directory of Open Access Journals (Sweden)

Sheng-Hsiung Chang

2015-04-01

Full Text Available This paper aims to incorporate the diffusion of innovation theory and conformity theory to explain consumers’ green purchase intentions. To this end, a conceptual model has been proposed and subjected to empirical verification with the use of a survey method. Using a sample of Taiwanese consumers who had the actual purchase experience of green detergents, this study employed structural equation modeling to verify the hypothesis proposed. The empirical results suggested that green viral communication was positively related to normative interpersonal influence, informational interpersonal influence and green purchase intention. Informational interpersonal influence also had a positive impact on green purchase intention. However, the relationship between consumer’s normative interpersonal influence and green purchase intention was not supported. Thus, this study concludes that green marketers must strengthen their green viral communications skills to enhance consumers’ purchase intentions. In addition, this study also contributes to the literature by stating that consumers’ susceptibility to informational interpersonal relationships is an important mediator in the green viral communication and green purchase intentions relationship. This study discusses implications of the findings and research limitations at the end of the paper.

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.

Pengaruh Green Practice Terhadap Green Consumer Behavior Di the Kemangi Restaurant, Hotel Santika Pandegiling Surabaya

OpenAIRE

Budiantoro, Anastasia Vianney; Irawan, Andrew; Kristanti, Monika; Aprilia, Adriana

2015-01-01

Penelitian ini dilakukan untuk mengetahui pengaruh green practice terhadap green consumer behavior di The Kemangi Restaurant. Teknik analisa yang digunakan dalam penelitian ini adalah kuantitatif dengan analisa regresi linier berganda. Penulis menggunakan 100 sampel untuk diteliti dengan melakukan survei kepada konsumen The Kemangi Restaurant. Hasil dari penelitian membuktikan bahwa ketiga variabel bebas berpengaruh positif. Namun, hanya variabel green donation yang memiliki pengaruh positif ...

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)

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)

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 ...

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

Into the Green Economy – Evolutionary Perspectives on Green Economic Change

DEFF Research Database (Denmark)

Andersen, Maj Munch

The recent ‘greening’ of the economy represents possible one of the most profound examples of economic change. While the environment used to be considered a burden to business this perspec-tive has changed making ‘eco-innovation’ increasingly recognized as a driver of economic devel-opment. Evolu......The recent ‘greening’ of the economy represents possible one of the most profound examples of economic change. While the environment used to be considered a burden to business this perspec-tive has changed making ‘eco-innovation’ increasingly recognized as a driver of economic devel...... of the greening of industry and the economy is of interest because of the focus on the fundamental social and economic difficulties of changing direction in technology. Defining the greening of the economy as a techno-economic paradigm change the paper suggests expanding on Perez’s framework (Perez, 1983, 2000...... problem solving, and simultaneously, the emergence of new green selection criteria on the market. These lead to a series of interrelated eco-innovations, which gain still more force as the green market matures. In the search for the origins of paradigmatic changes, the paper suggests to focus...

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.

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...

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

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

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…

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

Borrowing green : economic and environmental effects of green fiscal policy in the Netherlands

NARCIS (Netherlands)

Scholtens, B.

2001-01-01

This paper analyzes the economic and environmental impact of a policy instrument that is related to the tax deductibility of interest returns and dividend yields from specified 'green' projects. We investigate this so-called 'Green Project Facility' (Regeling Groenprojecten) in the Netherlands

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.

PENGARUH PENERAPAN GREEN ACCOUNTING TERHADAP KINERJA PERUSAHAAN

Directory of Open Access Journals (Sweden)

Hanifa Zulhaimi

2015-04-01

Full Text Available The purpose of this research is to analyze the implementation of green accounting and to find an impact of application of green accounting toward earning and stock price growth in Indonesian Industri. Industri activities oftentimes give some bad impact to environment surroundings such as natural devastation and the changes of culture, social and economic. Green accounting is a realization of corporate social responsibility to relieve the impact. The implementation of green accounting can give good image for the company however preliminary research found not many companies are implementing green accounting. This research will use quantitave approach and different test or  paired T-test will use for statistical testing, in order to test the research assumptions. Variable of this research are Green accounting, Earning per Shares and Stock Price Growth. This research is expected will contribute for the development of green accounting theory and enhancement of the implementation of green accounting especially in Indonesian Industri on Asian Economic community era.

Challenges of green chemistry in Ukraine

Directory of Open Access Journals (Sweden)

Shevtsova Ganna Ziyvna

2017-06-01

Full Text Available The article deals with study of Ukrainian chemical enterprises’ ecologisation issues and elaboration of the economic problems to realize principles of green chemistry. Theoretical aspects of green chemistry as a modern interdisciplinary conception, which reveals peculiarities to implement sustainable development paradigm in the chemical industry, are studied. Based on the analysis of essence and effectiveness to introduce international initiatives on sustainable development at the chemical industry enterprises, it is concluded that the implemented measures are only first steps on the way to realize key principles of green chemistry.It is proved that in order to promote conceptual ideas of the green chemistry further, it is reasonable to consider economic and marketing aspects of the ecological innovations: to provide economic effectiveness of green chemical products and technologies, to form ecological culture of consumption, to motivate green demand and to prevent market asymmetry of information.

GREEN GALAXIES IN THE COSMOS FIELD

Energy Technology Data Exchange (ETDEWEB)

Pan, Zhizheng; Kong, Xu; Fan, Lulu, E-mail: panzz@mail.ustc.edu.cn, E-mail: xkong@ustc.edu.cn [Center of Astrophysics, University of Science and Technology of China, Hefei 230026 (China)

2013-10-10

We present research on the morphologies, spectra, and environments of ≈2350 'green valley' galaxies at 0.2 < z < 1.0 in the COSMOS field. The bimodality of dust-corrected NUV–r {sup +} color is used to define 'green valley'; it removes dusty star-forming galaxies from galaxies that are truly transitioning between the blue cloud and the red sequence. Morphological parameters of green galaxies are intermediate between those of blue and red galaxy populations, both on the Gini-asymmetry and the Gini-M{sub 20} planes. Approximately 60%-70% of green disk galaxies have intermediate or big bulges, and only 5%-10% are pure disk systems, based on morphological classification using the Zurich Estimator of Structural Types. The obtained average spectra of green galaxies are intermediate between blue and red ones in terms of [O II], Hα, and Hβ emission lines. Stellar population synthesis on the average spectra shows that green galaxies are on average older than blue galaxies but younger than red galaxies. Green galaxies and blue galaxies have similar projected galaxy density (Σ{sub 10}) distributions at z > 0.7. At z < 0.7, the fractions of M{sub *} < 10{sup 10.0} M{sub ☉} green galaxies located in a dense environment are found to be significantly larger than those of blue galaxies. The morphological and spectral properties of green galaxies are consistent with the transitioning population between the blue cloud and the red sequence. The possible mechanisms for quenching star formation activities in green galaxies are discussed. The importance of active galactic nucleus feedback cannot be well constrained in our study. Finally, our findings suggest that environmental conditions, most likely starvation and harassment, significantly affect the transformation of M{sub *} < 10{sup 10.0} M{sub ☉} blue galaxies into red galaxies, especially at z < 0.5.

Green roof and storm water management policies: monitoring experiments on the ENPC Blue Green Wave

Science.gov (United States)

Versini, Pierre-Antoine; Gires, Auguste; Fitton, George; Tchiguirinskaia, Ioulia; Schertzer, Daniel

2015-04-01

Currently widespread in new urban projects, green roofs have shown a positive impact on urban runoff at the building/parcel scale. Nevertheless, there is no specific policy promoting their implementation neither in Europe nor in France. Moreover they are not taken into account (and usually considered as an impervious area) in the sizing of a retention basin for instance. An interesting example is located in the heart of the Paris-East Cluster for Science and Technology (Champs-sur-Marne, France). Since 2013 a large (1 ha) wavy-form vegetated roof (called bleu green wave) is implemented. Green roof area and impervious areas are connected to a large retention basin, which has been oversized. The blue green wave represents a pioneering site where an initially amenity (decorative) design project has been transformed into a research oriented one. Several measurement campaigns have been conducted to investigate and better understand the hydrological behaviour of such a structure. Rainfall, humidity, wind velocity, water content and temperature have been particularly studied. The data collected are used for several purposes: (i) characterize the spatio-temporal variability of the green roof response, (ii) calibrate and validate a specific model simulating its hydrological behavior. Based on monitoring and modeling results, green roof performances will be quantified. It will be possible to estimate how they can reduce stormwater runoff and how these performances can vary in space and in time depending on green roof configuration, rainfall event characteristics and antecedent conditions. These quantified impacts will be related to regulation rules established by stormwater managers in order to connect the parcel to the sewer network. In the particular case of the building of a retention basin, the integration of green roof in the sizing of the basin will be studied. This work is funded by the European Blue Green Dream project (http://bgd.org.uk/, funded by Climate

Organizational Green IT Adoption: Concept and Evidence

Directory of Open Access Journals (Sweden)

Qi Deng

2015-12-01

Full Text Available Green IT has emerged as an important research topic in information systems and in other areas, such as business sustainability management. Some progress has been made in our understandings of green IT in a wide area of research topics, ranging from the green IT definition to the motivation for adopting green IT by organizations. This paper provides a holistic review and explanation of why organizations adopt green IT. Based on an extensive review of extant studies and a broad theoretical foundation, the paper presents a theoretical framework on organizational green IT adoption (OGITA. For researchers, the study provides a comprehensive review of previous green IT adoption studies and a roadmap for future research. For practitioners, the study provides managers and policy makers a systematic analytical framework in guiding their business decisions.

The Influence of Legitimacy on a Proactive Green Orientation and Green Performance: A Study Based on Transitional Economy Scenarios in China

Directory of Open Access Journals (Sweden)

Baoshan Ge

2016-12-01

Full Text Available With environmental pollution, climate change and resource scarcity being serious global issues, green entrepreneurship is increasingly seen as an approach to simultaneously address economic performance, environmental impact and social responsibility. As green entrepreneurship needs to consider both venture performance and social responsibility, it will be subject to legitimacy constraints at the system level. Whether these legitimacy constraints are favorable to green enterprise is not yet clear from current research. Especially for transition economies, the problem of whether proactive green enterprises facing legitimacy constraints under institutional uncertainty can achieve green performance requires further study. Thus, a theoretical model to determine the relationship between green proactiveness orientation (GPO, green performance, legitimacy, and transitional economics was proposed. Based on the data from 235 new Chinese green firms, the empirical results suggest that green startups launch with a green proactiveness orientation, which enables them to acquire a green performance advantage over their competitors. Improvements in green performance is also shown to be driven by the pressure from institutional legitimacy. Better green performance can be easily achieved if green startups have a higher level of legitimacy. However, against the background of transitional economies, the increase in institutional uncertainty will damage the promotion of political legitimacy and make the enterprises that are subject to political legitimacy constraints lose their green performance. Currently, political legitimacy is no longer an impetus. However, the increase in institutional uncertainty will strengthen the promotion of commercial legitimacy and cause green-oriented startups to pursue more commercial interests. Thus, to a certain extent, it will lead to market uncertainty. The conclusion of this study not only provides guidance for startups in

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.

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....

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...

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

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:

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.

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.

7 CFR 51.1011 - Good green color.

Science.gov (United States)

2010-01-01

... 7 Agriculture 2 2010-01-01 2010-01-01 false Good green color. 51.1011 Section 51.1011 Agriculture... Standards for Persian (Tahiti) Limes Definitions § 51.1011 Good green color. Good green color means that the skin of the lime is of a good green color characteristic of the Persian variety. ...

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…

Analisis Green Product dan Green Marketing Strategy terhadap Keputusan Pembelian Produk The Body Shop di Manado Town Square

OpenAIRE

Soegoto, Agus Supandi; Lapian, Joyce; Ahmad, Fahlis

2016-01-01

: Pemasaran hijau (green marketing) akhir-akhir ini sudah merupakan trend di berbagai negara. Produk hijau (green product) semakin diminati karena pergeseran pola hidup dari produk yang instan ke produk yang menggunakan bahan alami dan baik untuk lingkungan. Untuk memenangkan persaingan Perusahaan harus mempunyai strategi yang tepat untuk memasarkan produknya. Penelitian ini bertujuan untuk mengetahui apakah Green Product dan Green Marketing Strategy secara simultan dan parsial apakah berpen...

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.

Green businesses in a clean energy economy: Analyzing drivers of green business growth in U.S. states

International Nuclear Information System (INIS)

Yi, Hongtao

2014-01-01

In a clean energy economy, green businesses play a central role by utilizing renewable energy technologies and employing green labor forces to provide clean energy services and goods. This paper aims at analyzing factors driving the growth and survival of green businesses in the U.S. states, with hypotheses proposed on the impacts from clean energy policies and tax incentives, labor market conditions, and economic and political environments. A fixed effect regression analysis is applied with a panel data set of 48 continental states from 1998 to 2007 in the United States. The statistical analysis with a longitudinal data set reveals that the adoption of renewable energy policies, the permission of renewable energy credits imports, the stringency of minimum wage legislations, and presence of clean energy business associations are the major driving forces of the green business development in the U.S. states. - Highlights: • This paper studies the growth of green businesses in the U.S. states. • The adoption of RPS (Renewable Portfolio Standard) is positively associated with number of green businesses. • Clean energy NGOs are positively associated with green business growth

Improving the urban green system and green network through the rehabilitation of railway rust areas

Directory of Open Access Journals (Sweden)

Hutter Dóra

2014-11-01

Full Text Available The Industrial Revolution had a negative impact on both the city and the environment. By the second half of the 19th century, the urban erosion of industrial cities cried for direct intervention and curing. The methods developed either along an urban or an anti-urban philosophy: they resulted in the new models of green belt systems aimed at solving all the main urban problems with restructuring the urban fabric, controlling the urban spread into the rural landscape, the lack of green areas and open spaces for recreation and social life, and the lack of green spaces for ventilation. Nowadays, the major cities and capitals around the globe are competing for titles such as healthier, more liveable or even greener city. Given the unfortunate attributes of the urban structure in the historical cities, the development of new transportation sites or green areas is an extremely difficult issue. On the other hand, in the big cities, the brownfield sites are considered as reserve areas for sustainable urban development. Reusing the brownfields and rust areas is already a land saving urban development approach and in case of a complex and ecological urban rehabilitation it can underlie the development of an efficient urban green system and green network.

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.

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)

Using Green Star Metrics to Optimize the Greenness of Literature Protocols for Syntheses

Science.gov (United States)

Duarte, Rita C. C.; Ribeiro, M. Gabriela T. C.; Machado, Adélio A. S. C.

2015-01-01

A procedure to improve the greenness of a synthesis, without performing laboratory work, using alternative protocols available in the literature is presented. The greenness evaluation involves the separate assessment of the different steps described in the available protocols--reaction, isolation, and purification--as well as the global process,…

Green competitiveness research on Chinese automotive enterprises

Directory of Open Access Journals (Sweden)

Yuanhui Li

2014-05-01

Full Text Available Purpose: More and more executives of automobileindustry in China start to recognize the concept of green competitiveness recently. However, relatively less research attention has been devoted to the consideration of measurement. This paper aims to find empirical approach to quantify green competitiveness for automotive enterprises. The connotation of green competitiveness is explored and one suite of evaluation index system has been proposed with four dimensions including environmental, resource, capability and knowledge.Design/methodology/approach: By introducing the factor analysis method, green competitiveness has been measured through an empirical analysis of 24 automotive enterprises within China.Findings: The results indicate that those elements, such as enterprise resource possession and utilization; environment, responsibility and knowledge; profitability; management efficiency, have significant effect on the green competitiveness for automotive enterprises. The further analysis also unveils the advantages and disadvantages of green competitiveness for each company and the direction for improvement.Research limitations/implications: Guide regulators and managers of automobile industry to take some measures to enhance their green competitive advantage.Practical implications: Provide practical methods to measure green competitiveness for automotive enterprises.Originality/value: This paper proposes an evaluation index system of green competitiveness for automotive enterprises. The suggestions of our research will be beneficial to enterprise executives and industry regulators.

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.