WorldWideScience

Sample records for premier ehec assay

  1. Enterohemorrhagic E. coli (EHEC) pathogenesis

    OpenAIRE

    Vanessa eSperandio; Y eNguyen

    2012-01-01

    Enterohemorrhagic Escherichia coli (EHEC) serotype O157:H7 is a human pathogen responsible for outbreaks of bloody diarrhea and hemolytic uremic syndrome (HUS) worldwide. Conventional antimicrobials trigger an SOS response in EHEC that promotes the release of the potent Shiga toxin that is responsible for much of the morbidity and mortality associated with EHEC infection. Cattle are a natural reservoir of EHEC, and approximately 75% of EHEC outbreaks are linked to the consumption of contamina...

  2. A Primary Test of EHEC O104:H4and EHEC O157:H7in Certain Kinds of Food in Wuhan,China%A Primary Test of EHEC O 104: H4 and EHEC O 157: H7 in Certain Kinds of Food in Wuhan, China

    Institute of Scientific and Technical Information of China (English)

    Wei Hechen; Xiong Yan

    2012-01-01

    Objective:This paper provided preliminary description of food contamination derived from Enterohemorrhagic Escherichia coli (EHEC)O104:H4 and EHEC O157:H7 in Wuhan in June 2011.Methods:47 food samples,including vegetables and meat,were subjectively sampled from some restaurants.PCR assays were used to detect EHEC O104:H4 and EHEC O157:H7.Results:The PCR results showed that none of the samples were positive for either EHEC O104:H4 or EHEC O 157:H7.Conclusion:Although large outbreaks of gastroenteritis and the hemolytic uremic syndrome caused by EHEC O104:H4 had occurred in some European countries,China has had few outbreaks associated with EHEC O104:H4.This shows that food supply is relatively safe in China.Nevertheless,many ongoing problems of food safety in China are still not solved showing the necessity of further studies on food safety.

  3. Enterohemorrhagic E. coli (EHEC pathogenesis

    Directory of Open Access Journals (Sweden)

    Vanessa eSperandio

    2012-07-01

    Full Text Available Enterohemorrhagic Escherichia coli (EHEC serotype O157:H7 is a human pathogen responsible for outbreaks of bloody diarrhea and hemolytic uremic syndrome worldwide. Conventional antimicrobials trigger an SOS response in EHEC that promotes the release of the potent Shiga toxin that is responsible for much of the morbidity and mortality associated with EHEC infection. Cattle are a natural reservoir of EHEC, and approximately 75% of EHEC outbreaks are linked to the consumption of contaminated bovine-derived products. This review will discuss how EHEC causes disease in humans but is asymptomatic in adult ruminants. It will also analyze factors utilized by EHEC as it travels through the bovine gastrointestinal tract that allow for its survival through the acidic environment of the distal stomachs, and for its ultimate colonization in the recto-anal junction. Understanding the factors crucial for EHEC survival and colonization in cattle will aid in the development of alternative strategies to prevent EHEC shedding into the environment and consequent human infection.

  4. Premier Hospital Historical Data

    Data.gov (United States)

    U.S. Department of Health & Human Services — To provide a historical overview of the participating hospitals, before the first project report, Premier Healthcare Informatics has used data already available for...

  5. PREMIER MEETS THE PRESS

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    On March 14, Premier Wen Jiabao addressed the Chinese and foreign media at a press conference after the closing meeting of the Third Session of the 11th National People’s Congress. Edited highlights on a number of economic and social issues follow:

  6. PREMIER MEETS THE PRESS

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    @@ March 14.Premier Wen Jiabao addressed the Chinese and foreign media at a press conference after the closing meeting of the Third Session of the 1 lth National People's Congress.Edited highlights on a number of economic and social issues follow:

  7. Microbiology: EHEC downregulates virulence in response to intestinal fucose.

    Science.gov (United States)

    Keeney, Kristie M; Finlay, B Brett

    2013-02-04

    Recent work has revealed that enterohaemorrhagic Escherichia coli encodes a two-component system, termed FusKR, which responds to fucose and represses expression of virulence genes. Furthermore, a representative member of the microbiota appears to cleave fucose from host glycans, indicating that the microbiota and EHEC may act in concert to suppress virulence gene expression.

  8. Premier Wen on Domestic Policies

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    The Second Session of the 11th National People’s Congress held a press conference at the Great Hall of the People on March 13. Premier Wen Jiabao answered questions from the Chinese and foreign press. Below are highlights of his answers on domestic policies.

  9. Our Measures Are Effective, Premier

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    @@ In an exclusive interview with Xinhua News Agency in Beijing on December 27, 2009, Chinese Premier Wen Jiabao shared his visions of the current economic situation in China and the government's economic policy orientations for 2010. Here are the highlights of the interview.

  10. Sequence Variations in the Flagellar Antigen Genes fliCH25 and fliCH28 of Escherichia coli and Their Use in Identification and Characterization of Enterohemorrhagic E. coli (EHEC O145:H25 and O145:H28.

    Directory of Open Access Journals (Sweden)

    Lothar Beutin

    Full Text Available Enterohemorrhagic E. coli (EHEC serogroup O145 is regarded as one of the major EHEC serogroups involved in severe infections in humans. EHEC O145 encompasses motile and non-motile strains of serotypes O145:H25 and O145:H28. Sequencing the fliC-genes associated with the flagellar antigens H25 and H28 revealed the genetic diversity of the fliCH25 and fliCH28 gene sequences in E. coli. Based on allele discrimination of these fliC-genes real-time PCR tests were designed for identification of EHEC O145:H25 and O145:H28. The fliCH25 genes present in O145:H25 were found to be very similar to those present in E. coli serogroups O2, O100, O165, O172 and O177 pointing to their common evolution but were different from fliCH25 genes of a multiple number of other E. coli serotypes. In a similar way, EHEC O145:H28 harbor a characteristic fliCH28 allele which, apart from EHEC O145:H28, was only found in enteropathogenic (EPEC O28:H28 strains that shared some common traits with EHEC O145:H28. The real time PCR-assays targeting these fliCH25[O145] and fliCH28[O145] alleles allow better characterization of EHEC O145:H25 and EHEC O145:H28. Evaluation of these PCR assays in spiked ready-to eat salad samples resulted in specific detection of both types of EHEC O145 strains even when low spiking levels of 1-10 cfu/g were used. Furthermore these PCR assays allowed identification of non-motile E. coli strains which are serologically not typable for their H-antigens. The combined use of O-antigen genotyping (O145wzy and detection of the respective fliCH25[O145] and fliCH28[O145] allele types contributes to improve identification and molecular serotyping of E. coli O145 isolates.

  11. Sequence Variations in the Flagellar Antigen Genes fliCH25 and fliCH28 of Escherichia coli and Their Use in Identification and Characterization of Enterohemorrhagic E. coli (EHEC) O145:H25 and O145:H28

    Science.gov (United States)

    Beutin, Lothar; Delannoy, Sabine; Fach, Patrick

    2015-01-01

    Enterohemorrhagic E. coli (EHEC) serogroup O145 is regarded as one of the major EHEC serogroups involved in severe infections in humans. EHEC O145 encompasses motile and non-motile strains of serotypes O145:H25 and O145:H28. Sequencing the fliC-genes associated with the flagellar antigens H25 and H28 revealed the genetic diversity of the fliCH25 and fliCH28 gene sequences in E. coli. Based on allele discrimination of these fliC-genes real-time PCR tests were designed for identification of EHEC O145:H25 and O145:H28. The fliCH25 genes present in O145:H25 were found to be very similar to those present in E. coli serogroups O2, O100, O165, O172 and O177 pointing to their common evolution but were different from fliCH25 genes of a multiple number of other E. coli serotypes. In a similar way, EHEC O145:H28 harbor a characteristic fliCH28 allele which, apart from EHEC O145:H28, was only found in enteropathogenic (EPEC) O28:H28 strains that shared some common traits with EHEC O145:H28. The real time PCR-assays targeting these fliCH25[O145] and fliCH28[O145] alleles allow better characterization of EHEC O145:H25 and EHEC O145:H28. Evaluation of these PCR assays in spiked ready-to eat salad samples resulted in specific detection of both types of EHEC O145 strains even when low spiking levels of 1–10 cfu/g were used. Furthermore these PCR assays allowed identification of non-motile E. coli strains which are serologically not typable for their H-antigens. The combined use of O-antigen genotyping (O145wzy) and detection of the respective fliCH25[O145] and fliCH28[O145] allele types contributes to improve identification and molecular serotyping of E. coli O145 isolates. PMID:26000885

  12. Sequence Variations in the Flagellar Antigen Genes fliCH25 and fliCH28 of Escherichia coli and Their Use in Identification and Characterization of Enterohemorrhagic E. coli (EHEC) O145:H25 and O145:H28.

    Science.gov (United States)

    Beutin, Lothar; Delannoy, Sabine; Fach, Patrick

    2015-01-01

    Enterohemorrhagic E. coli (EHEC) serogroup O145 is regarded as one of the major EHEC serogroups involved in severe infections in humans. EHEC O145 encompasses motile and non-motile strains of serotypes O145:H25 and O145:H28. Sequencing the fliC-genes associated with the flagellar antigens H25 and H28 revealed the genetic diversity of the fliCH25 and fliCH28 gene sequences in E. coli. Based on allele discrimination of these fliC-genes real-time PCR tests were designed for identification of EHEC O145:H25 and O145:H28. The fliCH25 genes present in O145:H25 were found to be very similar to those present in E. coli serogroups O2, O100, O165, O172 and O177 pointing to their common evolution but were different from fliCH25 genes of a multiple number of other E. coli serotypes. In a similar way, EHEC O145:H28 harbor a characteristic fliCH28 allele which, apart from EHEC O145:H28, was only found in enteropathogenic (EPEC) O28:H28 strains that shared some common traits with EHEC O145:H28. The real time PCR-assays targeting these fliCH25[O145] and fliCH28[O145] alleles allow better characterization of EHEC O145:H25 and EHEC O145:H28. Evaluation of these PCR assays in spiked ready-to eat salad samples resulted in specific detection of both types of EHEC O145 strains even when low spiking levels of 1-10 cfu/g were used. Furthermore these PCR assays allowed identification of non-motile E. coli strains which are serologically not typable for their H-antigens. The combined use of O-antigen genotyping (O145wzy) and detection of the respective fliCH25[O145] and fliCH28[O145] allele types contributes to improve identification and molecular serotyping of E. coli O145 isolates.

  13. Multiplexed Single Intact Cell Droplet Digital PCR (MuSIC ddPCR) Method for Specific Detection of Enterohemorrhagic E. coli (EHEC) in Food Enrichment Cultures

    Science.gov (United States)

    McMahon, Tanis C.; Blais, Burton W.; Wong, Alex; Carrillo, Catherine D.

    2017-01-01

    Foodborne illness attributed to enterohemorrhagic E. coli (EHEC), a highly pathogenic subset of Shiga toxin-producing E. coli (STEC), is increasingly recognized as a significant public health issue. Current microbiological methods for identification of EHEC in foods often use PCR-based approaches to screen enrichment broth cultures for characteristic gene markers [i.e., Shiga toxin (stx) and intimin (eae)]. However, false positives arise when complex food matrices, such as beef, contain mixtures of eae-negative STEC and eae-positive E. coli, but no EHEC with both markers in a single cell. To reduce false-positive detection of EHEC in food enrichment samples, a Multiplexed, Single Intact Cell droplet digital PCR (MuSIC ddPCR) assay capable of detecting the co-occurrence of the stx and eae genes in a single bacterial cell was developed. This method requires: (1) dispersal of intact bacteria into droplets; (2) release of genomic DNA (gDNA) by heat lysis; and (3) amplification and detection of genetic targets (stx and eae) using standard TaqMan chemistries with ddPCR. Performance of the method was tested with panels of EHEC and non-target E. coli. By determining the linkage (i.e., the proportion of droplets in which stx and eae targets were both amplified), samples containing EHEC (typically greater than 20% linkage) could be distinguished from samples containing mixtures of eae-negative STEC and eae-positive E. coli (0–2% linkage). The use of intact cells was necessary as this linkage was not observed with gDNA extracts. EHEC could be accurately identified in enrichment broth cultures containing excess amounts of background E. coli and in enrichment cultures derived from ground beef/pork and leafy-green produce samples. To our knowledge, this is the first report of dual-target detection in single bacterial cells using ddPCR. The application of MuSIC ddPCR to enrichment-culture screening would reduce false-positives, thereby improving the cost, speed, and accuracy of

  14. Precision Cleaning - Path to Premier

    Science.gov (United States)

    Mackler, Scott E.

    2008-01-01

    ITT Space Systems Division s new Precision Cleaning facility provides critical cleaning and packaging of aerospace flight hardware and optical payloads to meet customer performance requirements. The Precision Cleaning Path to Premier Project was a 2007 capital project and is a key element in the approved Premier Resource Management - Integrated Supply Chain Footprint Optimization Project. Formerly precision cleaning was located offsite in a leased building. A new facility equipped with modern precision cleaning equipment including advanced process analytical technology and improved capabilities was designed and built after outsourcing solutions were investigated and found lacking in ability to meet quality specifications and schedule needs. SSD cleans parts that can range in size from a single threaded fastener all the way up to large composite structures. Materials that can be processed include optics, composites, metals and various high performance coatings. We are required to provide verification to our customers that we have met their particulate and molecular cleanliness requirements and we have that analytical capability in this new facility. The new facility footprint is approximately half the size of the former leased operation and provides double the amount of throughput. Process improvements and new cleaning equipment are projected to increase 1st pass yield from 78% to 98% avoiding $300K+/yr in rework costs. Cost avoidance of $350K/yr will result from elimination of rent, IT services, transportation, and decreased utility costs. Savings due to reduced staff expected to net $4-500K/yr.

  15. Two novel EHEC/EAEC hybrid strains isolated from human infections.

    Directory of Open Access Journals (Sweden)

    Rita Prager

    Full Text Available The so far highest number of life-threatening hemolytic uremic syndrome was associated with a food-borne outbreak in 2011 in Germany which was caused by an enterohemorrhagic Escherichia coli (EHEC of the rare serotype O104:H4. Most importantly, the outbreak strain harbored genes characteristic of both EHEC and enteroaggregative E. coli (EAEC. Such strains have been described seldom but due to the combination of virulence genes show a high pathogenicity potential. To evaluate the importance of EHEC/EAEC hybrid strains in human disease, we analyzed the EHEC strain collection of the German National Reference Centre for Salmonella and other Bacterial Enteric Pathogens (NRC. After exclusion of O104:H4 EHEC/EAEC strains, out of about 2400 EHEC strains sent to NRC between 2008 and 2012, two strains exhibited both EHEC and EAEC marker genes, specifically were stx2 and aatA positive. Like the 2011 outbreak strain, one of the novel EHEC/EAEC harbored the Shiga toxin gene type stx2a. The strain was isolated from a patient with bloody diarrhea in 2010, was serotyped as O59:H-, belonged to MLST ST1136, and exhibited genes for type IV aggregative adherence fimbriae (AAF. The second strain was isolated from a patient with diarrhea in 2012, harbored stx2b, was typed as Orough:H-, and belonged to MLST ST26. Although the strain conferred the aggregative adherence phenotype, no known AAF genes corresponding to fimbrial types I to V were detected. In summary, EHEC/EAEC hybrid strains are currently rarely isolated from human disease cases in Germany and two novel EHEC/EAEC of rare serovars/MLST sequence types were characterized.

  16. Comparing balance and instep angle of premier and non- premier leg in female soccer athletics

    Directory of Open Access Journals (Sweden)

    Mahsa Mohammadzadeh

    2016-03-01

    Full Text Available This study is aimed to compare balanceand instep angle in premier and non- premier leg. The number of 30 female futsal athletics of Tehran Province (in the age of years old, in the weight of kg participated in this study. To determine premier and non- premier leg of subjects, they besides questioning requested to shoot 5 times and the leg with less error recorded as premier leg. Balance Error Scoring System (BESS test was used for evaluating static balance. 4 markers were used for evaluating instep angle according to Clark Model. For evaluating dynamic balance of subjects single-leg landing test on 3- axes force board with 1000 hertz frequency was used and the time to stability was computed in line with anteroposterior and internal- external based on sequential estimation technique. Data analyzing was done in two sections, first, instep angle was compared in different status and static and dynamic balance in premier and non- premier leg by paired t- test and then the relationship between instep angle was examined in different status with static balance (total score of balance in the test (BESS with instep angle in standing on two leg and single leg for premier leg and dynamic balance in 3 anteroposterior (AP, mediolateral (ML and vertical (V sides by using from correlation coefficient test (for normal data of Pearson correlation coefficient and Spearman abnormal data. There is significant difference between dynamic balance in premier and dynamic balance in non- premier leg of futsal athletics and the premier leg has further balance than non- premier leg. There is negative and significant relationship between dynamic balance and instep angle in all statusexcept premier leg of single- leg in vertical side. Futsal athletics in premier leg have further balance than non- premier leg.

  17. Sample processing for DNA chip array-based analysis of enterohemorrhagic Escherichia coli (EHEC

    Directory of Open Access Journals (Sweden)

    Enfors Sven-Olof

    2008-10-01

    Full Text Available Abstract Background Exploitation of DNA-based analyses of microbial pathogens, and especially simultaneous typing of several virulence-related genes in bacteria is becoming an important objective of public health these days. Results A procedure for sample processing for a confirmative analysis of enterohemorrhagic Escherichia coli (EHEC on a single colony with DNA chip array was developed and is reported here. The protocol includes application of fragmented genomic DNA from ultrasonicated colonies. The sample processing comprises first 2.5 min of ultrasonic treatment, DNA extraction (2×, and afterwards additional 5 min ultrasonication. Thus, the total sample preparation time for a confirmative analysis of EHEC is nearly 10 min. Additionally, bioinformatic revisions were performed in order to design PCR primers and array probes specific to most conservative regions of the EHEC-associated genes. Six strains with distinct pathogenic properties were selected for this study. At last, the EHEC chip array for a parallel and simultaneous detection of genes etpC-stx1-stx2-eae was designed and examined. This should permit to sense all currently accessible variants of the selected sequences in EHEC types and subtypes. Conclusion In order to implement the DNA chip array-based analysis for direct EHEC detection the sample processing was established in course of this work. However, this sample preparation mode may also be applied to other types of EHEC DNA-based sensing systems.

  18. Mastering Adobe Premiere Pro CS6

    CERN Document Server

    Ekert, Paul

    2013-01-01

    Designed to be practical and engaging, Mastering Adobe Premiere Pro CS6 is a project-based book to help you truly augment your skills and become a film editing hotshot.If you're just starting out or even migrating from existing video editing software, then this book is for you. With rapid progression through practical examples constructed to be both engaging and useful, Mastering Adobe Premiere Pro CS6 is ideal for learning the sometimes complex workflows of this powerful application.

  19. Evaluation of the SHIGA TOXIN QUIK CHEK and ImmunoCard STAT! EHEC as screening tools for the detection of Shiga toxin in fecal specimens.

    Science.gov (United States)

    Staples, Megan; Fang, Ning-Xia; Graham, Rikki Ma; Smith, Helen V; Jennison, Amy V

    2017-02-01

    In this study we evaluated the performance of the SHIGA TOXIN QUIK CHEK (Techlab®, Blacksburg, VA) and the ImmunoCard STAT! Enterohaemorrhagic E. coli (EHEC) (Meridian BioScience, Cincinnati, OH, USA) assays as methods for qualitatively detecting the presence of Shiga toxin in human fecal specimens. A multiplex PCR for the detection of stx1 and stx2 was used as the standard for comparison. The SHIGA TOXIN QUIK CHEK detected all known Shiga toxin subtypes with the exception of Stx2f, while the ImmunoCard STAT! EHEC was unable to identify four of the seven Stx2 subtypes, including Stx2b and Stx2d. When compared to multiplex PCR based on Shiga toxin gene presence alone both assays demonstrated 100% specificity, and gave sensitivity values of 50.0% and 41.2% respectively. Correlation between each assay and the multiplex PCR was calculated by the use of kappa, with both assays exhibiting a moderate level of agreement.

  20. Our Measures Are Effective,Premier

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    In an exclusive interview with Xinhua News Agency in Beijing on December 27,2009,Chinese Premier Wen Jiabao shared his visions of the current economic situation in China and the government’s economic policy orientations for 2010.Here are the highlights of the interview.

  1. Scottish Premier League Reading Stars Evaluation Report

    Science.gov (United States)

    National Literacy Trust, 2009

    2009-01-01

    Scottish Premier League (SPL) Reading Stars uses the motivational power of football to attract families who need support with literacy into a positive and friendly learning environment. It ran for the first time between March and August 2009 and attracted 225 children and 190 adults to take part in a series of inspirational learning sessions in 23…

  2. Cool Wool Iaunched at Premiere Vision

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    Visitors to Premiere Vision were greeted by the inspired The Woolmark Company image, rapidly becoming iconic, of a welcoming flock of stylish Merino sheep in sunglasses. It raised many a smile and set the scene for a feel- good exhibition, putting Cool Wool firmly at the doorway to the new summer season 2013.

  3. Pakistanis Eager to Welcome Premier Li Keqiang

    Institute of Scientific and Technical Information of China (English)

    Syed; Ali; Nawaz; Gilani

    2013-01-01

    <正>"We the Pakistani Nation,are very much eager to welcome Chinese Premier Li Keqiang on his first visit to Pakistan after taking over the Premiership of the People’s Republic of China whom we are enjoying deep rooted and exemplary friendship for

  4. Self-association of sodium deoxycholate with EHEC cellulose cooperatively induced by sodium dodecanoate.

    Science.gov (United States)

    Modolon, Samuel de M; Felippe, Arlindo C; Fizon, Tiago E; da Silva, Luciano; da Silva Paula, Marcos Marques; Dal-Bó, Alexandre G

    2014-10-13

    Some aspects of ethyl (hydroxyethyl) cellulose (EHEC) aqueous behavior in the presence of ionic surfactants are described in the literature; however, most of the studies reported address moderately concentrated solutions. Few studies have been carried out in the dilute regime using mixtures of biosurfactants. The main purpose of this work is to investigate the interaction of EHEC in the dilute regime and to verify the mixture of two surfactants: sodium deoxycholate (NaDC) and sodium dodecanoate (SDoD). The parameters of the surfactant to polymer association processes such as the critical aggregation concentration (cac) and saturation of the polymer by surfactants (psp) were determined from the plots of surface tension and specific conductivity versus surfactant concentration in basic conditions. The cmc of NaDC-SDoD mixtures showed non-ideal behavior. However, EHEC added to mixtures of SDoD and NaDC acts as a stabilizer for the mixed aggregate during the association process.

  5. Premier Wen hails sci-tech cooperation with CERN

    CERN Multimedia

    2004-01-01

    Premier Wen Jiabao met CERN's director general Dr Robert Aymar and physicist and Nobel laureate Dr Samuel Chao Chung Ting. Premier Wen emphasied the importance for China to collaborate on fundamental science (0.5 page)

  6. Survival of pathogenic enterohemorrhagic Escherichia coli (EHEC) and control with calcium oxide in frozen meat products.

    Science.gov (United States)

    Ro, Eun Young; Ko, Young Mi; Yoon, Ki Sun

    2015-08-01

    This study investigated both the level of microbial contamination and the presence of enterohemorrhagic Escherichia coli (EHEC) in frozen meat products, followed by the evaluation of its survival over 180 days under frozen temperature. We also examined the effect of calcium oxide on the populations of EHEC, E. coli O157:H7 and EPEC under both 10 °C and -18 °C storage conditions. Afterward, the morphological changes occurring in EHEC cells in response to freezer storage temperature and calcium oxide (CaO) treatments were examined using transmission electron microscopy. Among the frozen meat products tested, the highest contamination levels of total aerobic counts, coliforms and E. coli were observed in pork cutlets. Examination showed that 20% of the frozen meat products contained virulence genes, including verotoxin (VT) 1 and 2. Over 180 days of frozen storage and after 3 freeze-thaw cycles, the population of EHEC did not change regardless of the type of products or initial inoculated concentration, indicating the strong survival ability of EHEC. Subsequent testing revealed that the growth of three pathogenic E. coli strains was completely inhibited in meat patties prepared with 1% CaO, stored at 10 °C. However, the addition of 2% CaO was necessary to control the survival of EHEC, E. coli O157:H7 and EPEC in meat patties stored at -18 °C. CaO reduced the population of E. coli O157:H7 more effectively than the other EHEC and EPEC strains at both 10 °C and -18 °C. Transmission electron microscopy analysis revealed that exposed EHEC cells were resistant to the freezer storage temperature, although some cells incurred injury and death after several freeze-thaw cycles. Most of the cells exposed to CaO were found to have died or lost their cellular integrity and membranes, indicating that CaO has the potential to be used as a powerful antimicrobial agent for manufacturing frozen meat products. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Le Premier Amendement : un mythe

    Directory of Open Access Journals (Sweden)

    Claude‑Jean Bertrand

    2006-03-01

    Full Text Available Ce papier, plein de verve, d’humeur et d’humour, pose, comme sait si bien le faire l’auteur, des questions profondes sous une apparence paradoxale. Il a été présenté lors d’un colloque sur le Premier amendement organisé à l’Université Lumière-Lyon 2 les 17 et 18 janvier 2003. Certaines communications ont été publiées dans le volume XXIV, n°1 (2003, « Le premier amendement : un modèle américain des libertés » (sous la direction de Vincent Michelot de la Revue Tocqueville.

  8. MicroEnterprise Americas: Premiere Issue, 2001

    OpenAIRE

    Inter-American Development Bank (IDB)

    2001-01-01

    This premiere issue of MicroEnterprise Americas concentrates on the microfinance industry, a thriving segment of the Latin American financial sector that has rapidly expanded in the past five years. This issue explores looks at how market leaders have developed technologies, attracted investments, and developed tools for mitigating risk in the difficult financial climate of the past two years. MicroEnterprise Americas is published by the Inter-American Forum on Microenterprise, an annual even...

  9. MicroEnterprise Americas: Premiere Issue, 2001

    OpenAIRE

    Inter-American Development Bank (IDB)

    2001-01-01

    This premiere issue of MicroEnterprise Americas concentrates on the microfinance industry, a thriving segment of the Latin American financial sector that has rapidly expanded in the past five years. This issue explores looks at how market leaders have developed technologies, attracted investments, and developed tools for mitigating risk in the difficult financial climate of the past two years. MicroEnterprise Americas is published by the Inter-American Forum on Microenterprise, an annual even...

  10. [Pollution of EHEC O157:H7 in six types of food in Henan Province].

    Science.gov (United States)

    Zhang, Ding; Liao, Xingguang; Zhang, Xiuli; Li, Li

    2003-09-01

    In order to investigate the distribution of contamination of EHEC O157:H7 in 6 types of food in Henan Province and characterize the effect of seasonal factors on distribution of O157:H7 contamination so as to prevent and control food contamination, Henan Province was divided into five sampling areas according to the natural geographical features. Samples were taken randomly in summer and winter sales links. After selective culture of increasing reproduction of O157:H7, it was screened by immuno-gold reagent, and pathogenic bacterium were isolated after accumulation of immunomagnetic. Then, it was identified by the bioMérieux VITEK32 AMS system, GNI+ and sero-reaction. Results showed that 28 EHEC O157:H7 were isolated in 1463 samples among which the positive rate of raw meat and fresh vegetable was 3.3% and 3.2% respectively (the highest of all), and EHEC O157:H7 was not found in yogurt. The positive rate in summer (2.5%) was obviously higher than that in winter (1.1%). It could be concluded that the contamination of EHEC O157:H7 in food was serious. Its positive rate was positively correlated with the epidemic of infective diarrhea. Therefore, attention should be given by relevant departments concerned and spervision and testing should be strengthened in livestock and farmyard bird slaughtering link, vegetable production link and sales link for better prevention of the breaking out and epidemic of food born diseases.

  11. Construction and prokaryotic expression of the fusion protein Stx2B-IntiminC300 of EHEC O157:H7 and its immunoprophylactic potential

    Institute of Scientific and Technical Information of China (English)

    YONG YI; WEI JUN ZHANG; JIANG GU; PING LUO; XU HU MAO; WEN DE TONG; YING MA; MING ZEN; YONG HONG ZHU; QUAN MING ZOU; XIA AI; JIAN PING CHENG

    2006-01-01

    To construct and express the fusion protein Stx2B-IntiminC300 of EHEC O157: H7, and to further investigate its immunoprophylactic potential, the gene of Stx2B ( stx2b ) from EHEC O157: H7chromosome was cloned into pMD18-T vector. Thereafter, the amplified gene was cloned into prokaryotic expression plasmid pET-28a ( + )-eaeC300, which was constructed previously. The recombinant pasmid pET-28a( + )-stx2b-eaeC300 was transformed into E. coli BL21 (DE3). After inducement, the protein Stx2B-IntiminC300 was successfully expressed and analyzed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting and N-terminal amino acid residual sequencing. To evaluate its immunoprophylactic potential, it was primarily purified by ion-exchange chromatography and injected into 30 BALB/c mice with Al(OH)3 in the subscapular region. Ten days after the last booster vaccination, 20 mice were attacked with EHEC O157:H7 lysate and the protective efficacy was observed. In the present study, the gene of Stx2B-IntiminC300 was successfully cloned into pET-28a (+) vector. The results of SDS-PAGE and Western blotting assay showed that the fusion protein was successfully expressed in the inclusion body form, accounting for 25% of total expression products, and its molecular weight was about 43 kDa. The result of the N-terminal amino acid residual sequencing showed that it was identical to that of the molecular designed. The purity was about 75% after primary purification. Animal tests revealed that the fusion protein Stx2B-IntiminC300 has elicited high titer of protective antibody relatively. These results demonstrate that the fusion protein Stx2B-IntiminC300 is successfully expressed in prokaryotic expression system and shows certain immunoprophylactic potential.

  12. The QseC adrenergic signaling cascade in Enterohemorrhagic E. coli (EHEC.

    Directory of Open Access Journals (Sweden)

    David T Hughes

    2009-08-01

    Full Text Available The ability to respond to stress is at the core of an organism's survival. The hormones epinephrine and norepinephrine play a central role in stress responses in mammals, which require the synchronized interaction of the whole neuroendocrine system. Mammalian adrenergic receptors are G-coupled protein receptors (GPCRs; bacteria, however, sense these hormones through histidine sensor kinases (HKs. HKs autophosphorylate in response to signals and transfer this phosphate to response regulators (RRs. Two bacterial adrenergic receptors have been identified in EHEC, QseC and QseE, with QseE being downstream of QseC in this signaling cascade. Here we mapped the QseC signaling cascade in the deadly pathogen enterohemorrhagic E. coli (EHEC, which exploits this signaling system to promote disease. Through QseC, EHEC activates expression of metabolic, virulence and stress response genes, synchronizing the cell response to these stress hormones. Coordination of these responses is achieved by QseC phosphorylating three of the thirty-two EHEC RRs. The QseB RR, which is QseC's cognate RR, activates the flagella regulon which controls bacteria motility and chemotaxis. The QseF RR, which is also phosphorylated by the QseE adrenergic sensor, coordinates expression of virulence genes involved in formation of lesions in the intestinal epithelia by EHEC, and the bacterial SOS stress response. The third RR, KdpE, controls potassium uptake, osmolarity, and also the formation of lesions in the intestine. Adrenergic regulation of bacterial gene expression shares several parallels with mammalian adrenergic signaling having profound effects in the whole organism. Understanding adrenergic regulation of a bacterial cell is a powerful approach for studying the underlying mechanisms of stress and cellular survival.

  13. On the trail of EHEC/EAEC--unraveling the gene regulatory networks of human pathogenic Escherichia coli bacteria.

    Science.gov (United States)

    Pauling, Josch; Röttger, Richard; Neuner, Andreas; Salgado, Heladia; Collado-Vides, Julio; Kalaghatgi, Prabhav; Azevedo, Vasco; Tauch, Andreas; Pühler, Alfred; Baumbach, Jan

    2012-07-01

    Pathogenic Escherichia coli, such as Enterohemorrhagic E. coli (EHEC) and Enteroaggregative E. coli (EAEC), are globally widespread bacteria. Some may cause the hemolytic uremic syndrome (HUS). Varying strains cause epidemics all over the world. Recently, we observed an epidemic outbreak of a multi-resistant EHEC strain in Western Europe, mainly in Germany. The Robert Koch Institute reports >4300 infections and >50 deaths (July, 2011). Farmers lost several million EUR since the origin of infection was unclear. Here, we contribute to the currently ongoing research with a computer-aided study of EHEC transcriptional regulatory interactions, a network of genetic switches that control, for instance, pathogenicity, survival and reproduction of bacterial cells. Our strategy is to utilize knowledge of gene regulatory networks from the evolutionary relative E. coli K-12, a harmless strain mainly used for wet lab studies. In order to provide high-potential candidates for human pathogenic E. coli bacteria, such as EHEC, we developed the integrated online database and an analysis platform EhecRegNet. We utilize 3489 known regulations from E. coli K-12 for predictions of yet unknown gene regulatory interactions in 16 human pathogens. For these strains we predict 40,913 regulatory interactions. EhecRegNet is based on the identification of evolutionarily conserved regulatory sites within the DNA of the harmless E. coli K-12 and the pathogens. Identifying and characterizing EHEC's genetic control mechanism network on a large scale will allow for a better understanding of its survival and infection strategies. This will support the development of urgently needed new treatments. EhecRegNet is online via http://www.ehecregnet.de.

  14. Sample processing for DNA chip array-based analysis of enterohemorrhagic Escherichia coli (EHEC)

    OpenAIRE

    Enfors Sven-Olof; Wegrzyn Grzegorz; Basselet Pascal; Gabig-Ciminska Magdalena

    2008-01-01

    Abstract Background Exploitation of DNA-based analyses of microbial pathogens, and especially simultaneous typing of several virulence-related genes in bacteria is becoming an important objective of public health these days. Results A procedure for sample processing for a confirmative analysis of enterohemorrhagic Escherichia coli (EHEC) on a single colony with DNA chip array was developed and is reported here. The protocol includes application of fragmented genomic DNA from ultrasonicated co...

  15. Development of Digitex premier digital angiographic systems

    Energy Technology Data Exchange (ETDEWEB)

    Miura, Yoshiaki; Miura, Yusuke; Goto, Keiichi; Imanishi, Tetsuo; Miyamoto, Wataru [Shimadzu Corp., Medical Systems Division, Kyoto (Japan)

    2003-06-01

    The technique of interventional radiology has come to be widely utilized in the field of angiography. This has brought forth a strong demand that digital angiographic systems provide high efficiency in patient examinations and high level of interventional support. This report refers to our newly developed Digitex Premier Series digital angiographic systems, designed to meet the above demands. The new systems utilize a high-speed, wide-range C-arm system, a high-resolution image intensifier, a fluid-lubricant X-ray tube, and a digital image processing system, in order to ensure high patient examination efficiency. Their IVR (interventional radiology)-Master bed-side image controller further enhances the efficiency of patient examinations, and also, their CAT (comfortable angio terminal) and FMC (file management console) improve the patient examination throughput and diagnostic workflow of the systems. (author)

  16. [Identification of resistance and susceptibility to cefotaxime in EHEC O121 strains isolated from an outbreak at two nurseries].

    Science.gov (United States)

    Kikuchi, Koji; Ueno, Hiroyuki; Tomari, Kentaro; Kobori, Sumie; Kaetsu, Akihiko; Miyazaki, Motonobu

    2014-07-01

    A Shiga toxin 2 producing enterohemorrhagic Escherichia coli (EHEC) O121: H19 was isolated from a 2-year-old child who attending a nursery. An EHEC O121 outbreak in two nurseries (A, B), involving a total of 17 infected persons including 12 children, was revealed through contact investigation. The symptoms of all infected persons were almost all mild, and no one developed the hemolytic uremic syndrome. The combination use of desoxycholate-hydrogen sulfide-lactose (DHL) and CHROMagar STEC as selective isolation media was employed for efficient fecal examination. Nursery A and nursery B were combined as one group after the outbreak in nursery A was confirmed. As a result, EHEC O121 infected persons were also detected in children from nursery B. The 17 strains of EHEC O121 obtained from the total population showed almost the same pulsed-gel electrophoresis (PFGE) pattern, suggesting that these strains were very closely related. However, 13 of these 17 strains obtained from nursery A were susceptible to cefotaxime, whereas the remaining 4 strains obtained from nursery B showed cefotaxime resistance. A cefotaxime resistant Escherichia coli (E. coli) O86 strain was isolated in the stool specimen from a child who had been infected with the cefotaxime resistant EHEC O121. Both the cefotaxime resistant EHEC O121 and E. coli O86 had the same drug resistant gene (bla(CTX-M-1) group). The child was the index case of these 4 later cases and had received no antibiotics therapy prior to the laboratory examination. These findings suggested the possibility that an EHEC O121 strain had acquired a drug resistant gene from E. coli O86 in the digestive tract of the child.

  17. Fitness of Enterohemorrhagic Escherichia coli (EHEC)/Enteroaggregative E. coli O104:H4 in Comparison to That of EHEC O157: Survival Studies in Food and In Vitro.

    Science.gov (United States)

    Böhnlein, Christina; Kabisch, Jan; Meske, Diana; Franz, Charles M A P; Pichner, Rohtraud

    2016-11-01

    In 2011, one of the world's largest outbreaks of hemolytic-uremic syndrome (HUS) occurred, caused by a rare Escherichia coli serotype, O104:H4, that shared the virulence profiles of Shiga toxin-producing E. coli (STEC)/enterohemorrhagic E. coli (EHEC) and enteroaggregative E. coli (EAEC). The persistence and fitness factors of the highly virulent EHEC/EAEC O104:H4 strain, grown either in food or in vitro, were compared with those of E. coli O157 outbreak-associated strains. The log reduction rates of the different EHEC strains during the maturation of fermented sausages were not significantly different. Both the O157:NM and O104:H4 serotypes could be shown by qualitative enrichment to be present after 60 days of sausage storage. Moreover, the EHEC/EAEC O104:H4 strain appeared to be more viable than E. coli O157:H7 under conditions of decreased pH and in the presence of sodium nitrite. Analysis of specific EHEC strains in experiments with an EHEC inoculation cocktail showed a dominance of EHEC/EAEC O104:H4, which could be isolated from fermented sausages for 60 days. Inhibitory activities of EHEC/EAEC O104:H4 toward several E. coli strains, including serotype O157 strains, could be determined. Our study suggests that EHEC/EAEC O104:H4 is well adapted to the multiple adverse conditions occurring in fermented raw sausages. Therefore, it is strongly recommended that STEC strain cocktails composed of several serotypes, instead of E. coli O157:H7 alone, be used in food risk assessments. The enhanced persistence of EHEC/EAEC O104:H4 as a result of its robustness, as well as the production of bacteriocins, may account for its extraordinary virulence potential. In 2011, a severe outbreak caused by an EHEC/EAEC serovar O104:H4 strain led to many HUS sequelae. In this study, the persistence of the O104:H4 strain was compared with those of other outbreak-relevant STEC strains under conditions of fermented raw sausage production. Both O157:NM and O104:H4 strains could survive

  18. [ESBL-producing E. coli and EHEC in dogs and cats in the Tyrol as possible source of human infection].

    Science.gov (United States)

    Franiek, Natalie; Orth, Dorothea; Grif, Katharina; Ewers, Christa; Wieler, Lothar H; Thalhammer, Johann G; Würzner, Reinhard

    2012-01-01

    In contrast to infections with enterohaemorrhagic E. coli (EHEC), which are thought to be classical zoonosis, the zoonotic potential of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae is still widely unknown. The aim of our study was to determine the frequency of EHEC and ESBL-producing Enterobacteriaceae in domestic animals (dogs and cats) in the Tyrol. Among 228 fecal samples of dogs (n = 92) and cats (n = 136) three samples (1.3%) were positive in the EHEC-ELISA. In two of the three cases isolation of the organism was not possible, the third sample of a two-year-old crossbreed bitch yielded EHEC O103:H2. In twelve of 228 (5.3%) fecal samples 13 ESBL-producing Enterobacteriaceae (in ten cats and two dogs) were found.These animals mainly derived from homes for animals (ten animals, 83%). 75% of the isolates belonged to the CTX-M-1-group, 8% to the CTX-M-2-group and 17% to the CTX-M-9-group. One isolate was positive for CTX-M-1 and CTX-M-9. Typing of the 13 ESBL-producing isolates by multilocus sequence typing (MLST) showed ten different sequence types, which points out the importance of the horizontal transfer of mainly plasmid-coded ESBL genes. Transmission of EHEC and ESBL-producing Enterobacteriaceae from domestic animals to humans is possible, corroborated by the fact that the EHEC serotype found in one dog and the sequence types detected by MLST in several dogs and cats were previously reported to occur in severe human infection.

  19. Barriers to Trace-back in a Salad-associated EHEC Outbreak, Sweden, June 2013.

    Science.gov (United States)

    Edelstein, Michael; Sundborger, Camilla; Hergens, Maria-Pia; Ivarsson, Sofie; Dryselius, Rikard; Insulander, Mona; Jernberg, Cecilia; Hutin, Yvan; Wallensten, Anders

    2014-06-06

    In June-July 2013, six counties notified the Swedish Institute for Communicable Disease Control of enterohaemorrhagic E.coli (EHEC) infections among attendees at a hotel in Dalarna, Sweden. An outbreak control team investigated to identify the source and implement control measures. We included individuals who attended the hotel between June 19th-25th in a cohort. We asked them about animal contact, swimming, and consumption of food items during this time using a questionnaire. A confirmed case was an EHEC O157:H7 outbreak strain positive individual who developed abdominal pain or diarrhoea between June 20th-July 2nd. We described the outbreak in time, place and person, calculated risk ratios (RR) and 95% confidence intervals (CI). We investigated the kitchen, tested and traced back implicated food items. 172 individuals responded. We identified 19 confirmed cases (Median age: 17 years, 64% female) with symptom onset between June 22nd-27th. Eating green salad on June 20th was associated with illness (RR:3.7;CI:1.3-11). The kitchen mixed green salads without records and destroyed leftovers immediately. Hence we could not conduct trace-back or obtain microbiological confirmation. Green salad contaminated before entering the kitchen was the likely outbreak source. We recommended early collaboration with food agencies and better restaurant records to facilitate future investigations.

  20. Metaphor Analysis of Chinese Premier Wen’s Cambridge Speech

    Institute of Scientific and Technical Information of China (English)

    LUO Luo

    2014-01-01

    Metaphor is more than an ostensible decoration of language. It is an integral part of human thought of ideologized world. This article analyzes the metaphor use of Chinese Premier Wen Jiabao’s speech at Cambridge in February 2009, in an at-tempt to display how the preferred metaphors serve the purpose of this speech and reflect Premier Wen ’s construction of Chi-na’s situation.

  1. Molecular characterization of enterohemorrhagic Escherichia coli hemolysin gene (EHEC-hlyA)-harboring isolates from cattle reveals a diverse origin and hybrid diarrheagenic strains.

    Science.gov (United States)

    Askari Badouei, Mahdi; Morabito, Stefano; Najafifar, Arash; Mazandarani, Emad

    2016-04-01

    In the present study we investigated the occurrence of Escherichia coli strains harboring the gene encoding enterohemorrhagic E. coli hemolysin (EHEC-HlyA) in cattle and the association of this gene with various diarrheagenic E. coli (DEC) pathotypes. First, the bovine E. coli isolates were screened for EHEC-hlyA gene by PCR, and then they were characterized for the phylogenetic groups and the presence of the major virulence genes of different DEC pathotypes. In total, 25 virulence gene profiles were observed in 54 EHEC-hlyA+ isolates that reflect a considerable heterogeneity. The EHEC-hlyA+ strains were mostly associated with EHEC (72%), while only 7.4% were enteropathogenic E. coli (EPEC). We also showed the presence of estA gene of enterotoxigenic E. coli (ETEC) in 6 isolates (11.1%). Interestingly, two of the estA+ strains showed hybrid pathotypes with one carrying eae/estA (EPEC/ETEC), and the other one stx2/astA/estA (EHEC/ETEC). None of the isolates were related to enteroaggregative E. coli (EAggEC), enteroinvasive E. coli (EIEC), and necrotoxigenic E. coli (NTEC). The EHEC-plasmid encoded genes occurred in seven different combinations with EHEC-hlyA/saa/subA/espP being the most prevalent (46.3%). All stx-/eae+ strains carried O island 57 (OI-57) molecular marker(s) that may indicate these to be the progenitors of EHEC or strains losing stx. The most prevalent phylogroup was B1 (61.1%), but the most heterogeneous strains including the hybrid strains belonged to A phylogroup. Overall, our results indicate that cattle EHEC-hlyA encoding E. coli isolates consist of diverse diarrheagenic strains with the possible existence of hybrid pathotypes. Future studies are required to clarify the evolutionary aspects and clinical significance of these strains in humans and domestic animals.

  2. Premiere toob lavale jalgpallimeeskonna, inimkatsed ja punase tooli / Kairi Prints

    Index Scriptorium Estoniae

    Prints, Kairi, 1977-

    2012-01-01

    Premiere 2012 osalevad neli Eesti tantsukunstnikku: Svetlana Grigorjeva tantsulavastusega "sõp rus est", Kaisa Selde, Kristina-Maria Heinsalu ja Christin Lunts tantsulavastusega "fie", esmakordselt võtab osa välismaalane - sakslanna Mareike Franz tantsulavastusega "Duett". Kõik esietenduvad 9. veebruaril Kanuti gildi saalis

  3. Premiere toob lavale jalgpallimeeskonna, inimkatsed ja punase tooli / Kairi Prints

    Index Scriptorium Estoniae

    Prints, Kairi, 1977-

    2012-01-01

    Premiere 2012 osalevad neli Eesti tantsukunstnikku: Svetlana Grigorjeva tantsulavastusega "sõp rus est", Kaisa Selde, Kristina-Maria Heinsalu ja Christin Lunts tantsulavastusega "fie", esmakordselt võtab osa välismaalane - sakslanna Mareike Franz tantsulavastusega "Duett". Kõik esietenduvad 9. veebruaril Kanuti gildi saalis

  4. Open Access Publishing in Indian Premier Research Institutions

    Science.gov (United States)

    Bhat, Mohammad Hanief

    2009-01-01

    Introduction: Publishing research findings in open access journals is a means of enhancing visibility and consequently increasing the impact of publications. This study provides an overview of open access publishing in premier research institutes of India. Method: The publication output of each institution from 2003 to 2007 was ascertained through…

  5. Origin Detection During Food-borne Disease Outbreaks - A Case Study of the 2011 EHEC/HUS Outbreak in Germany.

    Science.gov (United States)

    Manitz, Juliane; Kneib, Thomas; Schlather, Martin; Helbing, Dirk; Brockmann, Dirk

    2014-04-01

    The key challenge during food-borne disease outbreaks, e.g. the 2011 EHEC/HUS outbreak in Germany, is the design of efficient mitigation strategies based on a timely identification of the outbreak's spatial origin. Standard public health procedures typically use case-control studies and tracings along food shipping chains. These methods are time-consuming and suffer from biased data collected slowly in patient interviews. Here we apply a recently developed, network-theoretical method to identify the spatial origin of food-borne disease outbreaks. Thereby, the network captures the transportation routes of contaminated foods. The technique only requires spatial information on case reports regularly collected by public health institutions and a model for the underlying food distribution network. The approach is based on the idea of replacing the conventional geographic distance with an effective distance that is derived from the topological structure of the underlying food distribution network. We show that this approach can efficiently identify most probable epicenters of food-borne disease outbreaks. We assess and discuss the method in the context of the 2011 EHEC epidemic. Based on plausible assumptions on the structure of the national food distribution network, the approach can correctly localize the origin of the 2011 German EHEC/HUS outbreak.

  6. [The EHEC O104:H4 outbreak in Germany 2011 - lessons learned?!].

    Science.gov (United States)

    Rissland, J; Kielstein, J T; Stark, K; Wichmann-Schauer, H; Stümpel, F; Pulz, M

    2013-04-01

    The EHEC O104:H4 outbreak 2011 in Germany provided numerous insights into the recognition and control of such epidemic situations. Food-borne outbreaks and their related dynamics may lead to a critical burden of disease and an eventual capacity overload of the medical care system. Possible difficulties in the microbiological diagnostics of new or significantly altered infectious agents may result in a delayed detection of the outbreak as well as the launching of interventional measures. Besides an early notification of the local public health office by the affected institutions, in which a complete electronic procedure and additional sentinel or surveillance instruments (e. g., in emergency departments of hospitals) may be of great help, an interdisciplinary cooperation of the local public health and food safety agencies is the key to an effective outbreak control. Corresponding organizations on the state and federal level should support the investigation process by microbiological diagnostics and advanced epidemiological analysis as well as examination of the food chains. Finally, successful crisis communication relies on "speaking with one voice" (not necessarily one person). Immediate, transparent, appropriate and honest information of the general public concerning the reasons, consequences and (counter-) measures of a crisis are the best means to keep the trust of the population and to counteract the otherwise inevitable speculations.

  7. Premiere of Film Nine-Mile Fragrance Held in Kenya

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    <正>The United Nations Environment Program (UNEP) and the United Nations Human Settlement Program (UN-HABITAT) in Kenya staged the premiere of the film Nine-Mile Fragrance depicting the Wenchuan earthquake relief efforts. The film, showing the unique culture of the Qiang ethnic group and the great spirit of love of the Chinese people in their earthquake relief work, left a deep impression on the African audience.

  8. Reevaluation of the Premier Clostridium difficile toxin A and B immunoassay with comparison to glutamate dehydrogenase common antigen testing evaluating Bartels cytotoxin and Prodesse ProGastro Cd polymerase chain reaction as confirmatory procedures.

    Science.gov (United States)

    Doing, Kirk M; Hintz, Marilyn S; Keefe, Calvin; Horne, Sarah; LeVasseur, Shelby; Kulikowski, Martha L

    2010-02-01

    Enzyme immunoassays are currently the most common tests used in the clinical laboratory for the detection of Clostridium difficile toxins; however, significant problems with their performance have recently been described. We prospectively reevaluated the Meridian Premier C. difficile toxin A/B assay with direct comparison to a 2-step algorithm that screened for C. difficile common antigen and compared cytotoxin and real-time polymerase chain reaction (PCR) as confirmatory procedures. The Premier assay lacked sufficient sensitivity, missing 25% of true-positive samples. PCR was the most sensitive method and the only procedure that allowed same day testing and reporting.

  9. Vice Premier Li Keqiang Meets Chinese and Russian Friendly Personages

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    <正>The scene: 5 pm on April 26 at the President Hotel in Moscow, and Vice Premier Li Keqiang, having just arrived in Russia for an official visit, cordially meets Chinese and Russian friendly personages from various sectors of society including people-to-people diplomatic, academic, cultural, scientific and educational, and business circles. Among them are experts and scholars in their seventies and eighties, important professionals of their respective workplaces in their prime, as well as students in the bloom of their youth.

  10. Lessons premier hospitals learned about implementing electronic health records.

    Science.gov (United States)

    DeVore, Susan D; Figlioli, Keith

    2010-04-01

    Implementing health information technology (IT) is a major strategic objective for providers. To pinpoint considerations that tie to success, the Premier health care alliance surveyed hospitals to develop an electronic health record best-practices library. Compiled from diverse health care organizations, the library outlines considerations to support "meaningful use" in the areas of computerized physician order entry, medication management, clinical documentation, reporting of measures, privacy, information exchange, management of populations' health, and personal health records. Best practices also uncovered strategies for securing executive leadership, culture change, communication, and support for clinicians. This paper summarizes lessons from the library, providing recommendations to speed up health IT implementation.

  11. Revisiting the STEC testing approach: using espK and espV to make enterohemorrhagic Escherichia coli (EHEC detection more reliable in beef

    Directory of Open Access Journals (Sweden)

    Sabine eDelannoy

    2016-01-01

    Full Text Available Current methods for screening Enterohemorrhagic Escherichia coli (EHEC O157 and non-O157 in beef enrichments typically rely on the molecular detection of stx, eae, and serogroup-specific wzx or wzy gene fragments. As these genetic markers can also be found in some non-EHEC strains, a number of ‘false positive’ results are obtained. Here, we explore the suitability of five novel molecular markers, espK, espV, ureD, Z2098, and CRISPRO26:H11 as candidates for a more accurate screening of EHEC strains of greater clinical significance in industrialized countries. Of the 1,739 beef enrichments tested, 180 were positive for both stx and eae genes. Ninety (50% of these tested negative for espK, espV, ureD, and Z2098, but twelve out of these negative samples were positive for the CRISPRO26:H11 gene marker specific for a newly emerging virulent EHEC O26:H11 French clone. We show that screening for stx, eae, espK, and espV, in association with the CRISPRO26:H11 marker is a better approach to narrow down the EHEC screening step in beef enrichments. The number of potentially positive samples was reduced by 48.88% by means of this alternative strategy compared to the European and American reference methods, thus substantially improving the discriminatory power of EHEC screening systems. This approach is in line with the EFSA (European Food Safety Authority opinion on pathogenic STEC published in 2013.

  12. EHEC O104:H4 in Germany 2011: Large outbreak of bloody diarrhea and haemolytic uraemic syndrome by shiga toxin-producing E. coli via contaminated food

    OpenAIRE

    2012-01-01

    In the summer of 2011 Germany experienced one of the largest outbreaks of a food-borne infection caused by enterohaemorrhagic Escherichia coli (EHEC) with the serotype O104:H4. A large number of cases with bloody diarrhea and haemolytic uraemic syndrome (HUS) occurred. Never before was such a high rate of HUS cases observed in an outbreak caused by a food-borne pathogen. The events in Germany caused by EHEC O104:H4 in the summer of 2011 show dramatically how rapidly an infectious agent ...

  13. Tears of a Chinese Premier Hussein Ismail Hussein

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    THE day Premier Wen took office he stated.""Leaders should be closer to the masses."" His visit to comfort and talk with everyday workers in Tongchuan City, Shaanxi Province on January 1, 2005, where a few weeks previously a gas blast in the Chenjiashan Coalmine had killed 166 workers, was by no means his first. But it was the first time he publicly shed tears. In one household that had lost its breadwinner, Wen Jiabao embraced the victim's son and shared his grief. He later had a simple lunch of steamed bread and tea in a tunnel 1,300 meters belowg round as he chatted with workers at another mine in the city.

  14. L’abri premier, de Vitruve à Nils-Udo

    Directory of Open Access Journals (Sweden)

    Nathalie Huvenne

    2011-05-01

    Full Text Available Cette étude se propose de mettre en écho les écrits de Vitruve, livre second, chapitre 1 du De architectura et les créations plastiques de l’artiste contemporain Nils-Udo.Ce vis-à-vis des réflexions de Vitruve avec celles de l’artiste nous permettra de tisser un lien entre ces deux hommes et de voir dans leurs travaux respectifs des points de similitude.Le texte extrait du De architectura II, 1, sous-titré par M. Nisard, De la manière de vivre des premiers hommes, et quels ont été les commenc...

  15. Premier Li Keqiang and Indian Prime Minister Modi Attend Regional Forum

    Institute of Scientific and Technical Information of China (English)

    Zhang; Min

    2015-01-01

    The First Forum of Leaders of the Regions of China and India,cosponsored by the CPAFFC and the China International Friendship Cities Association(CIFCA),was held at the Great Hall of the People in Beijing on May 15.Chinese Premier Li Keqiang and his Indian counterpart Narendra Modi gave addresses.Premier Li expressed his congrat-

  16. Effective Factors on Reducing the Number of Spectators in Iran Football Premier League

    Directory of Open Access Journals (Sweden)

    Amir Reza Khadem Azghadi

    2016-07-01

    Full Text Available Because of reducing the number of spectators of football premier league, this study is seeking for identifying factors put the most impact on this decline. The statistical population consisted of all spectators in Iran football premier league in 2015-16, out of which 395 spectators were randomly selected as the research samples. The data were collected via a researcher-made questionnaire. The first part of the questionnaire included demographic information and the second part, at 6 aspects, includes 35 questions analyzing the reasons for reducing the number of spectators in Iran's football Premier League. For analyzing data, it was used from first and second order confirmatory factor analysis based on structural equations through using SPSS 20 and LISREL 8.8 software. The results of first order confirmatory factor analysis showed that the measurement model of factors affecting on reducing the number of spectators of football premier league is an appropriate model and model parameters are significant. All factors are approved as effective variables on reducing the number of spectators of football premier league. The results also showed that the second order measurement model of effective factors on reducing the number of spectators of football premier league are also appropriate, and economic, facilitative, administrative, technical, cultural-social, and personal-family respectively put the most effects on reducing the number of spectators of football premier league. It is suggested for the sport marketers to analyze identified factors in this research and develop applicable strategies and guidelines for them.

  17. NE TIGER Premieres New Hua Fu At Bird’s Nest

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    On the evening of September 25, China’s leading luxury fashion brand NE TIGER presented its premiere fashion show of Hua Fu(Chinese national dress) at a concert of superstars from China, Japan and South Korea,

  18. Vaccination with DNA encoding truncated enterohemorrhagic Escherichia coli (EHEC factor for adherence-1 gene (efa-1’ confers protective immunity to mice infected with E. coli O157:H7

    Directory of Open Access Journals (Sweden)

    Roberto eRiquelme-Neira

    2016-01-01

    Full Text Available Enterohemorrhagic Escherichia coli (EHEC O157:H7 is the predominant causative agent of hemorrhagic colitis in humans and is the cause of haemolytic uraemic syndrome and other illnesses. Cattle have been implicated as the main reservoir of this organism. Here, we evaluated the immunogenicity and protective efficacy of a DNA vaccine encoding conserved sequences of truncated EHEC factor for adherence-1 (efa-1’ in a mouse model. Intranasal administration of plasmid DNA carrying the efa-1’ gene (pVAXefa-1’ into C57BL/6 mice elicited both humoral and cellular immune responses. In animals immunized with pVAXefa-1`, EHEC-secreted protein-specific IgM and IgG antibodies were detected in sera at day 45. Anti-EHEC-secreted protein sIgA was also detected in nasal and bronchoalveolar lavages. In addition, antigen-specific T-cell-proliferation, IL-10 and IFN-γ were observed upon re-stimulation with either heat-killed bacteria or EHEC-secreted proteins. Vaccinated animals were also protected against challenge with E. coli O157:H7 strain EDL933. These results suggest that DNA vaccine encoding efa-1´ have therapeutic potential in interventions against EHEC infections. This approach could lead to a new strategy in the production of vaccines that prevent infections in cattle.

  19. Vaccination with DNA Encoding Truncated Enterohemorrhagic Escherichia coli (EHEC) Factor for Adherence-1 Gene (efa-1') Confers Protective Immunity to Mice Infected with E. coli O157:H7.

    Science.gov (United States)

    Riquelme-Neira, Roberto; Rivera, Alejandra; Sáez, Darwin; Fernández, Pablo; Osorio, Gonzalo; del Canto, Felipe; Salazar, Juan C; Vidal, Roberto M; Oñate, Angel

    2015-01-01

    Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is the predominant causative agent of hemorrhagic colitis in humans and is the cause of haemolytic uraemic syndrome and other illnesses. Cattle have been implicated as the main reservoir of this organism. Here, we evaluated the immunogenicity and protective efficacy of a DNA vaccine encoding conserved sequences of truncated EHEC factor for adherence-1 (efa-1') in a mouse model. Intranasal administration of plasmid DNA carrying the efa-1' gene (pVAXefa-1') into C57BL/6 mice elicited both humoral and cellular immune responses. In animals immunized with pVAXefa-1', EHEC-secreted protein-specific IgM and IgG antibodies were detected in sera at day 45. Anti-EHEC-secreted protein sIgA was also detected in nasal and bronchoalveolar lavages. In addition, antigen-specific T-cell-proliferation, IL-10, and IFN-γ were observed upon re-stimulation with either heat-killed bacteria or EHEC-secreted proteins. Vaccinated animals were also protected against challenge with E. coli O157:H7 strain EDL933. These results suggest that DNA vaccine encoding efa-1' have therapeutic potential in interventions against EHEC infections. This approach could lead to a new strategy in the production of vaccines that prevent infections in cattle.

  20. Pro-Coagulant Endothelial Dysfunction Results from EHEC Shiga toxins and Host Damage-Associated Molecular Patterns

    Directory of Open Access Journals (Sweden)

    Chad L Mayer

    2015-04-01

    Full Text Available Hemolytic uremic syndrome (HUS from enterohemorrhagic E.coli infection (EHEC is a leading cause of kidney failure in otherwise healthy U.S. children. The bacterial Shiga toxins (Stx induce the characteristic coagulopathy of HUS, but the cell and organ damage during infection likely also releases inflammatory damage-associated molecular patterns (DAMPs that may exacerbate injury in concert with the toxins. To examine this, aortic and renal glomerular cell anticoagulant and barrier functions were studied after in vitro challenge with Stx1, Stx2 and DAMPs. There was significant loss of surface anticoagulant protein C pathway molecules, increased expression of prothrombotic PAR1 and reduced protein C activation capability by 15-27%. Histones nearly completely prevented activated protein C protection of endothelial cells from thrombin-induced permeability. In mice, lethal Stx2 challenge elevated plasma HMGB1 (day 2, 321±118%; p<0.01 and extracellular histones (day 3, 158±62%; p<0.01. Mice colonized with Stx2-expressing Citrobacter rodentium developed increased HMGB1 (day 5, 155±55%; p<0.01 and histones (day 3, 378±188%; p<0.01. Anti-histone antibody reduced both DAMPs to baseline, but was not sufficient to improve survival outcome or kidney function. Together, these data suggest a potential role for both Stx and DAMPs in producing endothelial injury and a prothrombotic environment.

  1. Genetic and mechanistic analyses of the periplasmic domain of the enterohemorrhagic E. coli (EHEC) QseC histidine sensor kinase.

    Science.gov (United States)

    Parker, Christopher T; Russell, Regan; Njoroge, Jacqueline W; Jimenez, Angel G; Taussig, Ron; Sperandio, Vanessa

    2017-01-30

    The histidine sensor kinase (HK) QseC, senses autoinducer-3 (AI-3), and the adrenergic hormones epinephrine and norepinephrine. Upon sensing these signals, QseC acts through three response regulators (RRs) to regulate expression of virulence genes in enterohemorrhagic E. coli (EHEC). The QseB, QseF and KdpE RRs that are phosphorylated by QseC constitute a tripartite signaling cascade having different and overlapping targets, including flagella and motility, the type three secretion system encoded by the locus of enterocyte effacement (LEE), and Shiga toxin. We modeled the tertiary structure of QseC's periplasmic sensing domain, and also aligned these sequences from 12 different species to identify the most conserved amino acids. We selected eight conserved aminoacids in all of these QseC homologs. These QseC site directed mutants were expressed and still able to autophosphorylate, albeit four mutants depicted increased basal level of phosphorylation. These mutants have differential flagella and motility, LEE and Shiga toxin expression phenotypes. We selected four mutants for more in depth analyses and found that they differed in their ability to phosphorylate QseB, KdpE and QseF. This suggests that these mutations in the periplasmic sensing domain affected the downstream of the QseC signaling cascade, and therefore, can influence which pathway QseC regulates.

  2. The 2014 presidential elections and their impact on the premier-presidential regime in Romania

    Directory of Open Access Journals (Sweden)

    Ionela Gavril

    2015-03-01

    Full Text Available First, we will demonstrate that, from an institutional perspective, Romania can labeled of premier-presidentialism regime, but the 2004 and 2009 elections have had a strong impact on the type of regime, meaning that several extra-constitutional factors led to the malfunction of the regime. Out of a total of 15 prime-minister nominations made after 1989, 8 can be considered deviations from the premier-presidential regime, their number being larger between 2004-2014 rather than in 1990-2000. The empirical analysis of the 2004-2014 period, highlighted three extra-constitutional factors that that made the premier-presidential regime be, in fact, a malfunctioning one: leadership style, crisis situations and the recent legitimacy of the president versus the parliament. By identifying the factors that influenced the regime type, we can determine some theoretical expectations following the 2014 elections. The success of a premier-presidentialism regime in Romania will be determined by the number of deviations from such a regime registered after the 2014 elections.

  3. Effectiveness of in-season manager changes in English Premier League Football

    NARCIS (Netherlands)

    Besters, Lucas; van Ours, Jan; van Tuijl, Martin

    2016-01-01

    We analyze the performance effects of in-season manager changes in English Premier League football during the seasons 2000/2001–2014/2015. We find that some managerial changes are successful, while others are counterproductive. On average, performance does not improve following a managerial replacem

  4. Vice Premier Zeng Peiyan Meets with Michael Cohrs, CEO of Corporate & Investment Banking of Deutsche Bank

    Institute of Scientific and Technical Information of China (English)

    2004-01-01

    <正>Invited by the CPAFFC, Michael Cohrs, Chief Executive Officer of the Corporate and Investment Banking of Deutsche Bank, visited Beijing from February 26 to 28, 2004. Vice Premier Zeng Peiyan of the State Council met with Cohrs and his party. Deutsche Bank AG, a global multipurpose bank

  5. Effectiveness of in-season manager changes in English Premier League Football

    NARCIS (Netherlands)

    Besters, Lucas; van Ours, Jan; van Tuijl, Martin

    We analyze the performance effects of in-season manager changes in English Premier League football during the seasons 2000/2001–2014/2015. We find that some managerial changes are successful, while others are counterproductive. On average, performance does not improve following a managerial

  6. Profil de l'etudiant du premier cycle des etudes medicales de Lome ...

    African Journals Online (AJOL)

    Profil de l'etudiant du premier cycle des etudes medicales de Lome et sa perception de l'enseignement de l'anatomie. ... Journal de la Recherche Scientifique de l'Universite de Lome ... aux différentes questions des paramètres étudiés.

  7. Premiere of TV Documentary Choe Chi-won Held in Beijing

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    <正>The premiere of the TV documentary Choe Chi-won was held in Beijing on August 19. The TV documentary,a joint production by the CPAFFC,the Jiangsu Provincial Association for Cultural Exchange with Foreign Countries,the

  8. High-intensity running in English FA Premier League soccer matches

    DEFF Research Database (Denmark)

    Bradley, Paul S.; Sheldon, William; Wooster, Blake

    2009-01-01

    The aims of this study were to (1) determine the activity profiles of a large sample of English FA Premier League soccer players and (2) examine high-intensity running during elite-standard soccer matches for players in various playing positions. Twenty-eight English FA Premier League games were.......01), attackers (2341 m, s=575, P game, high-intensity running distance was approximately 20% less than in the first 15-min period for wide midfielders (467 m, s=104 vs. 589 m, s=134, P ....01) and without ball possession (229 m, s=85 vs. 278 m, s=97, P game. Mean recovery time between very high-intensity running bouts was 72 s (s=28), with a 28% longer recovery time during the last 15 min than the first 15 min of the game (83 s, s=26 vs...

  9. Black Generation Y gender differences in Premier Soccer League spectator motives : sport marketing

    OpenAIRE

    T.E. Mofokeng; Bevan-Dye, A.L.

    2014-01-01

    The purpose of this study was to determine whether there are gender differences concerning Premier Soccer League (PSL) spectator motives amongst black Generation Y students in South Africa. In South Africa, the black Generation Y cohort (individuals born between 1986 and 2005) represents an important but under-researched market segment in that, in 2013, they made up 32 percent of the country's population. From a PSL marketing perspective, understanding the motives that drive game spectatorshi...

  10. Market segmentation in two-sided markets : tv rights for premier league

    OpenAIRE

    Kind, Hans Jarle; Sørgard, Lars

    2012-01-01

    This paper analyzes market segmentation in a two-sided market that consists of media consumers and advertisers. The analysis is motivated by a European Court of Justice Decision in October 2011, which allowed viewers to take advantage of international price differences and buy access to Premier League TV matches from whichever country they like. We compare complete market segmentation with the new situation where consumers can purchase from abroad (allowing for passive sales). Clearly, such a...

  11. ABOUT THE SMART SPORTS DEVELOPMENT. EVIDENCE FROM THE UK PREMIERE LEAGUE

    OpenAIRE

    Vlad Ionut Dumitrache

    2016-01-01

    Smart economy implies the development of key factors like global economy growth, competition, economic progress, economic prosperity, innovation. In the European top-level football, like the case of the British Premier League, financial indicators have demonstrated that the factors that define smart economy can be identified. The new rules of the financial fair-play policies and the ever growing revenues for television rights have created a new market in sports economy, one that identifies it...

  12. Logistique de transport pour le projet LHC enseignements des premiers secteurs

    CERN Document Server

    Prodon, S

    2003-01-01

    Ce papier dresse un premier bilan de la logistique de transport mise en place pour l'installation du LHC. Les moyens de planification mis en oeuvre seront tout d'abord évoqués avec notamment les réunions avec les groupes utilisateurs, l'élaboration de procédures de transport, la génération de listings d'articles à transporter ou encore l'établissement d'un planning des ressources. Cependant, les premiers travaux d'installation du LHC ont fait apparaître des divergences importantes entre le planning logistique établi et la réalité du terrain. Ces écarts seront analysés, qu'il s'agisse de différences sur le volume de matériel à acheminer, d'opérations non planifiées, de changements de plannings entraînant de longues et délicates traversées de chantiers ou de manque de planification des besoins en personnel dans certaines zones. Tous ces enseignements acquis au cours des premiers travaux devraient permettre de dégager des voies d'amélioration à mettre en place pour les prochains secteur...

  13. Vice Premier Zeng Peiyan Meets with Michael Cohrs,CEO of Corporate & Investment Banking of Deutsche Bank

    Institute of Scientific and Technical Information of China (English)

    LuHongwei

    2004-01-01

    Invited by the CPAFFC, Michael Cohrs, Chief Executive Officer of the Corporate and Investment Banking of Deutsche Bank, visited Beijing from February 26 to 28, 2004. Vice Premier Zeng Peiyan of the State Council met with Cohrs and his party.

  14. Premier Event

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    China and Russia look to move beyond resources and light manufacturing to hi-tech trade Russian Prime Minister Vladimir Putin paid an official visit to China at the invitation of his Chinese counterpart Wen Jiabao on October 11-12, Putin’s first visit abroad since he declared his intention to run for president next year.

  15. Premier Event

    Institute of Scientific and Technical Information of China (English)

    YU YAN

    2011-01-01

    Russian Prime Minister Vladimir Putin paid an official visit to China at the invitation of his Chinese counterpart Wen Jiabao on October 11-12,Putin's first visit abroad since he declared his intention to run for president next year.The visit strengthened diversified practical cooperation and promoted the SinoRussian strategic cooperative partnership in an all-around way,said Chinese analysts.

  16. Hubs and Authorities of the English Premier League for 2010-2011

    OpenAIRE

    Leznik, Michael

    2013-01-01

    In this work author applies well known web search algorithm Hyperlink - Induced Topic Search (HITS) to problem of ranking football teams in English Premier League (EPL). The algorithm allows the ranking of the teams using the notions of hubs and authorities well known for ranking pages in the World Wide Web. Results of the games introduced as a graph where losing team 'gives a link' to a winning team and, if draw registered both team give links to each other. In case of a win link is weighted...

  17. RANKING THE SPECTATORS’ DIFFICULTIES IN PURCHASING ELECTRONIC TICKETS OF FOOTBALL PREMIER LEAGUE

    Directory of Open Access Journals (Sweden)

    Ahmad Narimani

    2017-04-01

    Full Text Available This study aimed to rank the spectators’ difficulties in buying electronic tickets of football premier league matches at Azadi stadium. The population consisted of all spectators of Esteghlal-Persepolis match in the fifteenth league at Azadi stadium (N= 100000. According to Morgan table and using simple random sampling method, 500 participants were selected as sample. A researcher-made questionnaire was used for collecting the data; its face validity was confirmed by 15 experts and performing a pilot study on 30 subjects, its Cronbach’s alpha was calculated to be 0.86. Using SPSS 22, the descriptive and inferential (including Friedman test statistics was applied for analyzing the data. The findings showed that there was a significant difference between rankings of difficulties in buying electronic tickets of Football premier league matches at Azadi Stadium. The difficulties were ranked as: problem in ticket systems, early selling out of electronic tickets, lack of confidence to electronic ticket sale, lack of skill to work with the internet, low speed of internet, and lack of access to the internet

  18. The Relationship between Emotional Intelligence and Job Satisfaction among Coaches in Premier Under-20 Football League

    Directory of Open Access Journals (Sweden)

    Mehdi Moradi

    2012-06-01

    Full Text Available The purpose of present study was to examine the relationship between emotional intelligence and job satisfaction among coaches in premier Under-20 football league. The research method was descriptive-correlative, the performance method was survey, and data collection was done through field study. Research population consisted of 56 coaching staff in 14 teams participating in premier Under-20 football league. Finally, there were 48 questionnaires useable in data analysis. Emotional Intelligence Questionnaire (Syber Yashring and JDI (Wysocki & Kromm were used to collect the data. Descriptive statistics was used to describe data, Kolmogorov-Smirnov test was used to know whether the distribution of data was normal, and Pearson correlation and stepwise regression were applied to investigate the significance of hypotheses. Results showed that there was significant association between emotional intelligence, subscale self-awareness, subscale empathy, and subscale social skills with job satisfaction (p≤0.05. However, there was not significant association between subscale self-motivation and subscale self-control with job satisfaction. Self-awareness, empathy, and social skills (predictors predicted job satisfaction (criterion significantly. Predicted value of self-awareness, empathy, and social skills was 0.4, 0.29, and 0.26 respectively. Training and aging increase emotional intelligence so it is predicted more job satisfaction over the time. From other side, clubs and football federation as the head can create scientific atmosphere and instruct psychological coaching principles. It will lead to enjoy creative, willing players as output.

  19. ABOUT THE SMART SPORTS DEVELOPMENT. EVIDENCE FROM THE UK PREMIERE LEAGUE

    Directory of Open Access Journals (Sweden)

    Vlad Ionut Dumitrache

    2016-11-01

    Full Text Available Smart economy implies the development of key factors like global economy growth, competition, economic progress, economic prosperity, innovation. In the European top-level football, like the case of the British Premier League, financial indicators have demonstrated that the factors that define smart economy can be identified. The new rules of the financial fair-play policies and the ever growing revenues for television rights have created a new market in sports economy, one that identifies itself with the criteria identifies in studies regarding smart economy. This paper comparatively examines the determinants of four indicators of the football team quality in the British Premier League, in order to find out whether a common set of potential determinants could be effective in improving all four indicators of quality, without worsening any of them. This allows finding what measures undertaken at the level of football teams could raise the football team quality. Considering the subjective and multidimensional nature of the football team quality, we first propose four indicators that might be appropriate to define this latent summative measure. Then we select a number of four potentially common determinants of the football team quality, and finally discuss the empirical results, based on panel generalized least squares regression models. The television broadcasting rights are found to be the most important determinant of the football team quality.

  20. Study on rapid method for detecting Enterohemorrhagic Escherichia coil O104 in fruits and vegetables%快速检测果蔬中肠出血性大肠杆菌EHEC O104方法研究

    Institute of Scientific and Technical Information of China (English)

    李晓虹; 吴悦

    2013-01-01

    目的:针对德国暴发流行的肠出血性大肠杆菌EHEC O104并结合我国的特点,建立一套科学的快速检测和鉴定果蔬中肠出血性大肠杆菌EHEC O104的快速检测方法.方法:利用ELISA方法作为快速初筛方法,结合传统生化鉴定及PCR检测作为鉴定方法.结果:经特异性和灵敏度检测,方法特异性达到100%,灵敏度达到1.0×101 CFU/ml.对市售的50个果蔬样品进行检测,均未检出.结论:利用该方法建立的快速检测果蔬中肠出血性大肠杆菌EHEC:O104标准方法,可以广泛应用在果蔬中的快速检测.%Objective: According to the outbreak of EHEC 0104 strain in Germany, to establish a rapid method for detection of Enterohemorrhagic Escherichia coli 0104 in fruits and vegetables. Methods: ELISA was used for rapid preliminary screening, the conventional biochemical identification method and PCR were employed for identification. Results: With these above methods, the specificity was 100%, the sensitivity was 1.0 × 10 CFU/ml. Fifty fruit and vegetable samples were all negative for Enterohemorrhagic escherichia coli 0104.Conclusion: This method can be widely used for the detection of Enterohemorrhagic Escherichia coli 0104 in fruits and vegetables.

  1. Future perspectives, applications and challenges of genomic epidemiology studies for food-borne pathogens: A case study of Enterohemorrhagic Escherichia coli (EHEC) of the O157:H7 serotype.

    Science.gov (United States)

    Eppinger, Mark; Cebula, Thomas A

    2015-01-01

    The shiga-toxin (Stx)-producing human pathogen Escherichia coli serotype O157:H7 is a highly pathogenic subgroup of Stx-producing E. coli (STEC) with food-borne etiology and bovine reservoir. Each year in the U. S., approximately 100,000 patients are infected with enterohemorrhagic E. coli (EHEC) of the O157:H7 serotype. This food-borne pathogen is a global public health threat responsible for widespread outbreaks of human disease. Since its initial discovery in 1982, O157:H7 has rapidly become the dominant EHEC serotype in North America. Hospitalization rates among patients as high as 50% have been reported for severe outbreaks of human disease. Symptoms of disease can rapidly deteriorate and progress to life-threatening complications such as Hemolytic Uremic Syndrome (HUS), the leading cause of kidney failure in children, or Hemorrhagic Colitis. In depth understanding of the genomic diversity that exists among currently circulating EHEC populations has broad applications for improved molecular-guided biosurveillance, outbreak preparedness, diagnostic risk assessment, and development of alternative toxin-suppressing therapeutics.

  2. The highly virulent 2006 Norwegian EHEC O103:H25 outbreak strain is related to the 2011 German O104:H4 outbreak strain.

    Directory of Open Access Journals (Sweden)

    Trine M L'Abée-Lund

    Full Text Available In 2006, a severe foodborne EHEC outbreak occured in Norway. Seventeen cases were recorded and the HUS frequency was 60%. The causative strain, Esherichia coli O103:H25, is considered to be particularly virulent. Sequencing of the outbreak strain revealed resemblance to the 2011 German outbreak strain E. coli O104:H4, both in genome and Shiga toxin 2-encoding (Stx2 phage sequence. The nucleotide identity between the Stx2 phages from the Norwegian and German outbreak strains was 90%. During the 2006 outbreak, stx(2-positive O103:H25 E. coli was isolated from two patients. All the other outbreak associated isolates, including all food isolates, were stx-negative, and carried a different phage replacing the Stx2 phage. This phage was of similar size to the Stx2 phage, but had a distinctive early phage region and no stx gene. The sequence of the early region of this phage was not retrieved from the bacterial host genome, and the origin of the phage is unknown. The contaminated food most likely contained a mixture of E. coli O103:H25 cells with either one of the phages.

  3. Description sémantique de dans un premier temps : de la composition syntagmatique au discours

    Directory of Open Access Journals (Sweden)

    Catherine Schnedecker

    2011-12-01

    Full Text Available Cet article est consacré à l’expression adverbiale réputée sérielle dans un premier temps. Y est d’abord décrit le comportement sémantique de la locution dont le sens reste, pour une bonne partie, compositionnel, étant donné l’importance de la dimension temporelle et les contraintes qu’elle fait peser sur la nature des constituants de la configuration. Ensuite, nous montrons que l’expression dans un premier temps n’est « sérielle » que dans une partie de ses emplois. Dans les autres, non signalés dans la littérature, dans un premier temps marque une relation d’altérité qui peut être « faible » (i. e. encore teintée de temporalité ou « forte » (i. e. nettement contradictoire.The aim of this paper is to describe the semantic behavior of the French expression dans un premier temps, known as a serial marker. We show how its meaning is partly compositional, given the importance of the temporal dimension and the constraints which this places on the nature of the constituents. We then show that dans un premier temps is “serial” in only some of its uses. In others, not reported in the literature, dans un premier temps expresses a relation of “otherness” which can be either “weak” (i.e. keeping a temporal value, or “strong” (i.e. sharply contradictory.

  4. Les premiers textes de René Thom sur la morphogenèse et la linguistique : 1966-1970.

    OpenAIRE

    Petitot, Jean

    2015-01-01

    Au milieu des années 1960, René Thom commença à rédiger ses premiers textes sur les applications à la morphogenèse en biologie et à la syntaxe actantielle en linguistique de la théorie des déploiements universels de singularités de fonctions differentiables et de la stabilité structurelle. Cette note présente et commente ses cinq premiers articles dans ces domaines.

  5. Sur la loi de répartition du k-ième facteur premier d'un entier

    Science.gov (United States)

    de Koninck, J.-M.; Tenenbaum, G.

    2002-09-01

    Soit {pk(n)}w(n)k=1 la suite croissante des facteurs premiers distincts d'un entier n. Nous donnons, lorsque k [rightward arrow] [infty infinity], une approximation uniforme de la loi de répartition limite de la fonction arithmétique n [mapsto A:] pk(n), précisant ainsi un résultat classique d'Erdos. Deux applications en sont déduites, relatives à la médiane de cette loi et à celle de la fonction « nombre de facteurs premiers ».

  6. Vers une mélancolie des premiers romans ?

    OpenAIRE

    2017-01-01

    Johan Faerber :Je vais peut-être tout d’abord présenter brièvement ton premier roman, Le Black Note afin de t’interroger sur les questions rétrospectives qu’il suscite en toi et chercher ainsi à apercevoir ta propre lecture de ce moment inaugural de ton œuvre. Il s’agit, on le sait, du récit d’un groupe de jazz amateur taraudé par un modèle écrasant, celui de John Coltrane. Ce groupe sombre progressivement dans la drogue et disparaît à la mort de son leader, Paul, brûlé vif dans l’incendie de...

  7. Berlo Janet C. et Ruth B. Phillips, Amérique du Nord. Arts premiers

    OpenAIRE

    Mauzé, Marie

    2014-01-01

    Avec Amérique du Nord. Arts premiers signé de deux historiennes de l’art, Janet Berlo et Ruth Phillips, nous avons là un ouvrage qui fait partie d’une nouvelle génération de travaux sur les arts nord-amérindiens, à l’instar du livre de David W. Penney, North American Indian Art, publié en 2004. Connues pour l’excellence de leurs recherches, Berlo et Phillips proposent une étude exhaustive sur l’ensemble des arts visuels nord-amérindiens considérés dans leur contexte culturel et historique. Da...

  8. Un premier apprentissage mathématique, la construction du nombre

    OpenAIRE

    Incerti, Estelle; Migy, Pierre

    2014-01-01

    Ce travail de mémoire professionnel se penche sur la question d’un premier apprentissage mathématique fondamental, celui de la construction du nombre. Cette recherche tend à comprendre le rôle que porte l’enseignant au niveau de cet apprentissage dans les premières années de la scolarité d’un élève. Elle met en lumière les éléments théoriques nécessaires à l’acquisition du concept de nombre et les paramètres auxquels il faut prêter tout particulièrement attention en tant qu’enseignant. Au tra...

  9. [Sõltumatu Tantsu Ühenduse poolt korraldatud sarjast "Premiere"] / Evelin Lagle ; küsinud Tambet Kaugema

    Index Scriptorium Estoniae

    Lagle, Evelin, 1986-

    2012-01-01

    Uutele koreograafidele pühendatud sarja "Premiere" programmis osalevad tantsulavastustega neli tantsukunstnikku Eestist - Tallinna Ülikooli lõpetanud Svetlana Grigorjeva, Turu Kunstiakadeemia lõpetanud Kaisa Selde, Viljandi Kultuuriakadeemia lõpetanud Kristina-Maria Heinsalu ja Tallinna Ülikooli lõpetanud Christin Lunts

  10. Qualitative Impact Assessment 2010: An Independent Study Conducted by BDRC Continental, Ltd., February-July 2010. Premier League Reading Stars

    Science.gov (United States)

    National Literacy Trust, 2010

    2010-01-01

    Premier League Reading Stars (PLRS) is in its eighth year. To complement a pre-post quantitative survey, an impact evidence base was required to inform consideration of continued funding into 2011 and beyond. PLRS is very highly regarded among child participants, parents, and librarians. The structure of the scheme, its basis on football, and the…

  11. [Sõltumatu Tantsu Ühenduse poolt korraldatud sarjast "Premiere"] / Evelin Lagle ; küsinud Tambet Kaugema

    Index Scriptorium Estoniae

    Lagle, Evelin, 1986-

    2012-01-01

    Uutele koreograafidele pühendatud sarja "Premiere" programmis osalevad tantsulavastustega neli tantsukunstnikku Eestist - Tallinna Ülikooli lõpetanud Svetlana Grigorjeva, Turu Kunstiakadeemia lõpetanud Kaisa Selde, Viljandi Kultuuriakadeemia lõpetanud Kristina-Maria Heinsalu ja Tallinna Ülikooli lõpetanud Christin Lunts

  12. The effect of playing formation on high-intensity running and technical profiles in English FA Premier League soccer matches

    DEFF Research Database (Denmark)

    Bradley, Paul S; Carling, Chris; Archer, Dave

    2011-01-01

    The aim of this study was to examine the effect of playing formation on high-intensity running and technical performance during elite soccer matches. Twenty English FA Premier League games were analysed using a multiple-camera computerized tracking system (n = 153 players). Overall ball possessio...

  13. Enzyme assays.

    Science.gov (United States)

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

    2009-01-07

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

  14. Predicting Football Matches Results using Bayesian Networks for English Premier League (EPL)

    Science.gov (United States)

    Razali, Nazim; Mustapha, Aida; Yatim, Faiz Ahmad; Aziz, Ruhaya Ab

    2017-08-01

    The issues of modeling asscoiation football prediction model has become increasingly popular in the last few years and many different approaches of prediction models have been proposed with the point of evaluating the attributes that lead a football team to lose, draw or win the match. There are three types of approaches has been considered for predicting football matches results which include statistical approaches, machine learning approaches and Bayesian approaches. Lately, many studies regarding football prediction models has been produced using Bayesian approaches. This paper proposes a Bayesian Networks (BNs) to predict the results of football matches in term of home win (H), away win (A) and draw (D). The English Premier League (EPL) for three seasons of 2010-2011, 2011-2012 and 2012-2013 has been selected and reviewed. K-fold cross validation has been used for testing the accuracy of prediction model. The required information about the football data is sourced from a legitimate site at http://www.football-data.co.uk. BNs achieved predictive accuracy of 75.09% in average across three seasons. It is hoped that the results could be used as the benchmark output for future research in predicting football matches results.

  15. Evaluation of sports nutrition knowledge of New Zealand premier club rugby coaches.

    Science.gov (United States)

    Zinn, Caryn; Schofield, Grant; Wall, Clare

    2006-04-01

    Little is known about if and how team coaches disseminate nutrition information to athletes. In a census survey, New Zealand premier rugby coaches (n = 168) completed a psychometrically validated questionnaire, received by either Internet or standard mail (response rate, 46%), identifying their nutrition advice dissemination practices to players, their level of nutrition knowledge, and the factors determining this level of knowledge. The majority of coaches provided advice to their players (83.8%). Coaches responded correctly to 55.6% of all knowledge questions. An independent t-test showed coaches who imparted nutrition advice obtained a significantly greater score, 56.8%, than those not imparting advice, 48.4% (P = 0.008). One-way ANOVA showed significant relationships between total knowledge score of all coaches and qualifications [F(1,166) = 5.28, P = 0.001], own knowledge rating [F(3,164) = 6.88, P = 0.001] and nutrition training [F(1,166) = 9.83, P = 0.002]. We conclude that these rugby coaches were inadequately prepared to impart nutrition advice to athletes and could benefit from further nutrition training.

  16. Strategies for injury prevention in Brazilian football: Perceptions of physiotherapists and practices of premier league teams.

    Science.gov (United States)

    Meurer, Maurício Couto; Silva, Marcelo Faria; Baroni, Bruno Manfredini

    2017-07-25

    To describe the physiotherapists perceptions and the current practices for injury prevention in elite football (soccer) clubs in Brazil. Cross-sectional study. Group of Science in Sports & Exercise, Federal University of Healthy Sciences of Porto Alegre (Brazil). 16 of the 20 football clubs involved in the Brazilian premier league 2015. Physiotherapists answered a structured questionnaire. Most physiotherapists (∼88%) were active in design, testing and application of prevention programs. Previous injury, muscle imbalance, fatigue, hydration, fitness, diet, sleep/rest and age were considered "very important" or "important" injury risk factors by all respondents. The methods most commonly used to detect athletes' injury risk were: monitoring of biochemical markers (100% of teams), isokinetic dynamometry (81%), questionnaires (75%), functional movement screen (56%), fleximetry (56%) and horizontal jump tests (50%). All clubs used strength training, functional training, core exercises and balance/proprioception exercises in their injury prevention program; and Nordic hamstring exercise and other eccentric exercises were used by 94% of clubs. "FIFA 11+" prevention program was adapted by 88% of clubs. Physiotherapists perceptions and current practices of injury prevention within Brazilian elite football clubs were similar to those employed in developed countries. There remains a gap between clinical practice and scientific evidence in high performance football. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. The influence of situational variables on ball possession in the English Premier League.

    Science.gov (United States)

    Bradley, Paul Simon; Lago-Peñas, Carlos; Rey, Ezequiel; Sampaio, Jaime

    2014-12-01

    Abstract The aims of this study were twofold: (1) to examine the influence of situational variables on ball possession in elite soccer and (2) to quantify the variables that discriminate between high or low percentage ball possession teams (HPBPT and LPBPT) across different playing positions. Match performance data were collected from English Premier League matches using a multiple-camera system. Data were examined using linear regression, a 2 × 5 factorial analysis of variance and discriminant analysis. Playing against weak opposition was associated with an increase (P variables (P variables that discriminated performance between HPBPT and LPBPT were different for various playing positions, although the number of successful passes was the most common discriminating variable. The results demonstrate that HPBPT and LPBPT developed different possession strategies during matches and that selected variables such as successful passes were identified to explain these data trends across various playing positions. Combinations of variables could be used to develop a probabilistic model for predicting time spent in possession by teams.

  18. Technical Performance Analysis of Iran Premier League Soccer Players in 2012-2013 Season

    Directory of Open Access Journals (Sweden)

    Mohsen Javani

    2015-10-01

    Full Text Available Background and purpose of study : analysis of IRAN premier league soccer players’ technical performance in season 2012-2013, using a computerized match analysis system (Borhan Mobin Development Management Co, IRAN. Material and methods: in this study, data were obtained from 120 players, who performed in competitions 90 minutes. The players were classified into 3 positional roles: defenders, midfielders and forwards. Technical performance variables analysis included: total passes, total successful passes, pass accuracy, total shots; total shots to target, shot accuracy, ball interception and ball losses. The data were statistically analyzed by one-way ANOVA, Kruskal-Wallis, Mann-Whitney U and Tukey post hoc test. Results : The findings of this study showed that players performed about 45 passes per competition. Midfielders and defenders had significantly higher number of passes than forwards. Pass accuracy was about 67% and there were no significant differences between positional roles. Also, the players performed about 0.8 shots per competition, forwards and midfielders had significantly higher number of shots than defenders. Shot accuracy was about 31%; midfielders and forwards had significantly higher shot accuracy than defenders. Forwards showed significantly lower ball interception and higher ball losses than other positions. Conclusion : The result of this study showed that there were significant differences between some technical actions in positional roles. Therefore, coaches can use this information for individualization of training according to playing positions and for optimization of training in the amateur game.

  19. Development of a one-step PCR assay with nine primer pairs for the detection of five diarrheagenic Escherichia coli types.

    Science.gov (United States)

    Oh, Kyung-Hwan; Kim, Soo-Bok; Park, Mi-Sun; Cho, Seung-Hak

    2014-06-28

    Certain Escherichia coli (E. coli) strains have the ability to cause diarrheal disease. Five types of diarrheagenic E. coli have been identified, including EHEC, ETEC, EPEC, EAEC, and EIEC. To detect these five diarrheagenic types rapidly, we developed a one-step multiplex PCR (MPPCR) assay using nine primer pairs to amplify nine virulence genes specific to the different virotypes, with each group being represented (i.e., stx1 and stx2 for EHEC, lt, sth, and stp for ETEC, eaeA and bfpA for EPEC, aggR for EAEC, and ipaH for EIEC). The PCR primers were constructed using MultAlin. The sensitivity and specificity of the constructed multiplex PCR primers were measured using DNA isolated from diarrheagenic E. coli strains representing each group. The limits of detection were as follows: 5 × 10(1) CFU/ml for EHEC, 5 × 10(3) CFU/ml for ETEC expressing lt and sth, 5 × 10(4) CFU/ml for ETEC expressing stp, 5 × 102 CFU/ml for EPEC, 5 × 10(4) CFU/ml for EAEC, and 5 × 10(2) CFU/ml for EIEC. To confirm the specificity, C. jejuni, C. perfringens, S. Typhimurium, V. parahaemolyticus, L. monocytogenes, Y. enterocolitica, B. cereus, and S. aureus were used as negative controls, and no amplification was obtained for these. Moreover, this kit was validated using 100 fecal samples from patients with diarrhea and 150 diarrheagenic E. coli strains isolated in Korea. In conclusion, the multiplex PCR assay developed in this study is very useful for the rapid and specific detection of five diarrheagenic E. coli types. This single-step assay will be useful as a rapid and economical method, as it reduces the cost and time required for the identification of diarrheagenic E. coli.

  20. Peat moss fuels R and D: Premier Tech is using sphagnum peat moss to develop all kinds of sophisticated products

    Energy Technology Data Exchange (ETDEWEB)

    Desiront, A.

    2004-10-01

    Development by Premier Tech, a Quebec multinational company, of a unique system for recycling construction waste using peat moss as the starting material, is reported. Premier Tech enjoys a high profile in sectors such as the life sciences, wastewater treatment systems, industrial packaging and a variety of other fields not readily associated with peat moss. The best known products of the company, the Allegro line of growth media designed to promote greenhouse growing, Biomax compost and the Promix plant growth media, all based on research on mycorrhizal association, and the development strategy guiding the company's operations, are described briefly. The company's Ecoflo and Ecoprocess filters for treating domestic and municipal wastewater, both of which played significant roles in recovering evidence from the rubble of the World Trade Center after September 11, 2001, are highlighted.

  1. Chinese Dream——Concert in Commemoration of 115th Birth Anniversary of Premier Zhou Enlai Held

    Institute of Scientific and Technical Information of China (English)

    Our; Staff; Reporter

    2013-01-01

    <正>The theme song of the film The Founding of a Republic sung by male vocalists Dai Yuqiang and Wei Song reverberated in the Opera Hall at the National Center for the Performing Arts on the `evening of March 14. It marked the start of the concert in commemoration of the 115th anniversary of the birth of Premier Zhou Enlai, with "Chinese Dream" as the theme.

  2. Philibert Delorme’s Divine Proportions and the Composition of the Premier Tome de l’Architecture

    Directory of Open Access Journals (Sweden)

    Sara Galletti

    2014-06-01

    Full Text Available In his 'Premier tome de l’architecture' (1567 — the first original, comprehensive architectural treatise written by a French author — Philibert Delorme (c. 1514–1570 claims to be the first to formulate a theory of divine proportions, which he describes as a set of rules recorded in the Old Testament as directly dictated by God to men for the construction of the Ark of Noah, the Ark of the Covenant, and the Temple and House of Solomon. Yet the author does not develop the theory of divine proportions in the 'Premier tome' and postpones it instead to the second volume of his treatise. As a second volume was never published (and likely never written, Delorme readers are left with a handful of less-than-coherent references and illustrations of a theory that remains largely obscure. Yet the elements of theory of divine proportions contained in the 'Premier tome' provide historians with an understanding of the genesis of the treatise itself, thus ultimately helping to raise broader questions about the book and its author. This paper shows how Delorme’s divine proportions offer a key to understanding the conception and composition of his treatise as well as to the process of intellectual development of the author and the changes in the nature and scope of his written work.

  3. Identificación de péptidos bloqueantes del sistema de secreción de tipo III y de la adherencia a epitelios de Escherichia coli enterohemorrágico (EHEC) y Escherichia coli enteropatógeno (EPEC)

    OpenAIRE

    Larzábal, Mariano

    2010-01-01

    Escherichia coli Enteropatógeno (EPEC) y Escherichia coli Enterohemorrágico (EHEC) se encuentran asociadas con diarrea en seres humanos. EPEC es una de las principales causas de diarrea infantil en países desarrollados, mientras que EHEC es responsable de enfermedades cuyo espectro clínico incluye diarrea, colitis hemorrágica y síndrome urémico hemolítico (SUH), que es la principal causa de insuficiencia renal aguda de niños menores de 5 años en la Argentina. La expresión de la toxina Shiga (...

  4. Premier Wen Jiabao Chaired a State Council Executive Meeting to Review and Deploy the Program to Further Implement the Restructuring and Reinvigoration Plan for Key Industries

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    @@ Premier Wen Jiabao chaired a State Council executive meeting to review and deploy the program to further implement the restructuring and reinvigoration plan for key industries on February 24th,2010.

  5. Corporate social responsibility and mental health: the Premier League football Imagine Your Goals programme.

    Science.gov (United States)

    Henderson, Claire; O'Hara, Stefanie; Thornicroft, Graham; Webber, Martin

    2014-08-01

    Football is increasingly used to facilitate recovery in mental health services, often in partnership with football clubs. However, few clubs have made mental health part of their corporate social responsibility programmes until recently. We report the impact on participants of the 'Imagine Your Goals' programme, run by 16 Premier League football clubs in conjunction with England's Time to Change programme to reduce mental health-related stigma and discrimination. Mixed methods evaluation used pre/post measures of well-being, access to social capital, focus groups held early on and towards the end of the two-year programmes, and questionnaires for coaching staff. There were no significant changes to participants' mental well-being scores between baseline and follow-up, nor to the total number of social resources accessible through their networks. However, there was a statistically significant increase at follow-up in the mean score of the personal skills subscale of the Resource Generator-UK. Participants' individual skills were also higher at follow-up. Qualitative data showed programmes had largely met participants' expectations in terms of socializing, providing structure and improving fitness levels, exceeded expectations in relationships with coaching staff and additional activities, but did not always meet them in improving football skills. Participants varied in their knowledge of exit opportunities, depending on which club's programme they attended. A minority of clubs reported difficulties in recruitment and concerns about planning for the future of the projects. Football clubs and the charitable foundations they set up can successfully deliver programmes to people with mental health problems which improve access to personal skills social capital and have other potential benefits.

  6. Descriptive epidemiology of injuries in a Brazilian premier league soccer team

    Directory of Open Access Journals (Sweden)

    Fachina RJ

    2013-06-01

    Full Text Available Rafael Júlio de Freitas Guina Fachina,1,2 Marília dos Santos Andrade,3 Fernando Roberto Silva,4 Silas Waszczuk-Junior,4 Paulo César Montagner,1 João Paulo Borin,1 Claudio Andre Barbosa de Lira5 1Departamento de Ciência do Esporte, Faculdade de Educação Física, Universidade Estadual de Campinas (UNICAMP, Campinas, Brazil; 2Confederação Brasileira de Basketball (CBB, Rio de Janeiro, Brazil; 3Departamento de Fisiologia, Universidade Federal de São Paulo, São Paulo, Brazil; 4Grêmio Barueri Futebol LTDA, Barueri, Brazil; 5Setor de Fisiologia Humana e do Exercício, Universidade Federal de Goiás, Câmpus Jataí, Jataí, Brazil Abstract: Soccer, which has a large number of participants, has a high injury incidence that causes both financial and time burdens. Therefore, knowledge about the epidemiology of soccer injuries could allow sports-medicine professionals, such as physicians and physiotherapists, to direct their work in specific preventive programs. Thus, our aim was to conduct an epidemiological survey of injuries sustained by professional soccer players from the same team who participated in the Brazilian championship premier league in 2009. To this end, we evaluated retrospectively player medical records from the team, which included name, date of birth, position, date of injury, mechanism of injury, and type of injury. In the period of study, 95 injuries were recorded: 42 (44.2% were recorded during matches, and 53 (55.8% during the training period. Injuries occurred more frequently in midfielders and strikers. All injuries happened in the lower limb, most of the injuries were muscular, and most occurred as the result of collisions with other athletes. In summary, this study demonstrates that there is a need for greater safety awareness in the training environment. Keywords: injuries, epidemiology, soccer players

  7. Hospital arrival time and functional outcome after acute ischaemic stroke: results from the PREMIER study.

    Science.gov (United States)

    León-Jiménez, C; Ruiz-Sandoval, J L; Chiquete, E; Vega-Arroyo, M; Arauz, A; Murillo-Bonilla, L M; Ochoa-Guzmán, A; Carrillo-Loza, K; Ramos-Moreno, A; Barinagarrementeria, F; Cantú-Brito, C

    2014-05-01

    Information regarding hospital arrival times after acute ischaemic stroke (AIS) has mainly been gathered from countries with specialised stroke units. Little data from emerging nations is available. We aim to identify factors associated with achieving hospital arrival times of less than 1, 3, and 6 hours, and analyse how arrival times are related to functional outcomes after AIS. We analysed data from patients with AIS included in the PREMIER study (Primer Registro Mexicano de Isquemia Cerebral) which defined time from symptom onset to hospital arrival. The functional prognosis at 30 days and at 3, 6, and 12 months was evaluated using the modified Rankin Scale. Among 1096 patients with AIS, 61 (6%) arrived in <1 hour, 250 (23%) in <3 hours, and 464 (42%) in <6 hours. The factors associated with very early (<1 hour) arrival were family history of ischemic heart disease and personal history of migraines; in <3 hours: age 40-69 years, family history of hypertension, personal history of dyslipidaemia and ischaemic heart disease, and care in a private hospital; in <6 hours: migraine, previous stroke, ischaemic heart disease, care in a private hospital, and family history of hypertension. Delayed hospital arrival was associated with lacunar stroke and alcoholism. Only 2.4% of patients underwent thrombolysis. Regardless of whether or not thrombolysis was performed, arrival time in <3 hours was associated with lower mortality at 3 and 6 months, and with fewer in-hospital complications. A high percentage of patients had short hospital arrival times; however, less than 3% underwent thrombolysis. Although many factors were associated with early hospital arrival, it is a priority to identify in-hospital barriers to performing thrombolysis. Copyright © 2013 Sociedad Española de Neurología. Published by Elsevier Espana. All rights reserved.

  8. PREMIERS RESULTATS SUR LES COMPORTEMENTS DE SUBSISTANCE SOLUTREENS A LA GROTTE ROCHEFORT (MAYENNE, FRANCE

    Directory of Open Access Journals (Sweden)

    Céline Bemilli

    2013-09-01

    Full Text Available Le site de la grotte Rochefort (Mayenne, France a livré depuis 2005 plusieurs centaines de restes fauniques issus des couches Solutréennes. Le Renne et le Cheval en sont les principales composantes et reflètent le fond commun du régime alimentaire des Solutréens. Nous présenterons ici les premiers résultats de l’analyse archéozoologique concernant l’exploitation de ces deux principaux taxons. Plusieurs recollages ont été réalisés. L’exceptionnelle conservation des restes permet, outre une excellente lecture des traces de découpe, une première reconstitution des modalités d’acquisition et d’exploitation des animaux, ainsi que leur saison d’abattage. La rareté des ensembles fauniques solutréens fouillés et étudiés récemment donne à celui de la grotte Rochefort un impact particulier et inédit.Several hundreds faunal remains were discovered since 2005 in the Solutrean‘s level at Rochefort cave (Mayenne, France. Reindeer and Horse are the main taxa identified and are the baseline of the Solutrean diet. We present here the preliminary results of the faunal analysis and discuss the exploitation of these two main taxa. A number of refits were possible. At the same time, the exceptional preservation of the artefacts together with well-defined cut marks allows to reconstruct procurement and use patterns of these animals as well as the season they were killed. Recently excavated and analysed Solutrean faunal assemblages are rare which make the yet unreported Rochefort cave faunal assemblage even more important and unprecedented.

  9. Angiogenesis Assays.

    Science.gov (United States)

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

    2016-01-01

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

  10. Evaluation of a new real-time PCR assay for the direct detection of diarrheagenic Escherichia coli in stool specimens.

    Science.gov (United States)

    Eigner, U; Hiergeist, A; Veldenzer, A; Rohlfs, M; Schwarz, R; Holfelder, M

    2017-02-01

    Diarrheagenic E. coli (DEC) are one of the most common causes for diarrhea worldwide, especially in children. We evaluated the rapid RIDA ® GENE (RG) real-time multiplex PCR assays (R-Biopharm, Darmstadt, Germany) for the detection of the most important diarrheagenic E. coli. Three hundred fifteen liquid or non-formed stool specimens were examined. The results of the RG multiplex assays were compared to specific PCR methods. The sensitivity and specificity of the RG PCRs were as follows, 100%/100% for the detection of EHEC, 96.3% and 99% for EPEC, 100% and 100% for the detection of EAEC, ETEC and EIEC, respectively. Overall, the RG real-time PCR system for the detection of DEC tested in this study provided reliable and rapid results and shows the ability as a useful addendum for the detection of diarrheagenic E. coli in the medical laboratory.

  11. Comparison of five assays for detection of Clostridium difficile toxin.

    Science.gov (United States)

    Chapin, Kimberle C; Dickenson, Roberta A; Wu, Fongman; Andrea, Sarah B

    2011-07-01

    Performance characteristics of five assays for detection of Clostridium difficile toxin were compared using fresh stool samples from patients with C. difficile infection (CDI). Assays were performed simultaneously and according to the manufacturers' instructions. Patients were included in the study if they exhibited clinical symptoms consistent with CDI. Nonmolecular assays included glutamate dehydrogenase antigen tests, with positive findings followed by the Premier Toxin A and B Enzyme Immunoassay (GDH/EIA), and the C. Diff Quik Chek Complete test. Molecular assays (PCR) included the BD GeneOhm Cdiff Assay, the Xpert C. difficile test, and the ProGastro Cd assay. Specimens were considered true positive if results were positive in two or more assays. For each method, the Youden index was calculated and cost-effectiveness was analyzed. Of 81 patients evaluated, 26 (32.1%) were positive for CDI. Sensitivity of the BD GeneOhm Cdiff assay, the Xpert C. difficile test, the ProGastro Cd assay, C. Diff Quik Chek Complete test, and two-step GDH/EIA was 96.2%, 96.2%, 88.5%, 61.5%, and 42.3%, respectively. Specificity of the Xpert C. difficile test was 96.4%, and for the other four assays was 100%. Compared with nonmolecular methods, molecular methods detected 34.7% more positive specimens. Assessment of performance characteristics and cost-effectiveness demonstrated that the BD GeneOhm Cdiff assay yielded the best results. While costly, the Xpert C. difficile test required limited processing and yielded rapid results. Because of discordant results, specimen processing, and extraction equipment requirements, the ProGastro Cd assay was the least favored molecular assay. The GDH/EIA method lacked sufficient sensitivity to be recommended. Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

  12. Autoassociação de misturas dos Surfactantes Dodecanoato de Sódio (SDoD e Decanoato de Sódio (SDeC com o Polímero Hidrofobicamente Modificado Etil(HidroxietilCelulose (EHEC

    Directory of Open Access Journals (Sweden)

    Alexandre G. Dal-Bó

    2011-01-01

    Full Text Available In this work, the interactions between the non-ionic polymer of ethyl(hydroxyethylcellulose (EHEC and mixed anionic surfactant sodium dodecanoate (SDoD-sodium decanoate (SDeC in aqueous media, at pH 9.2 (20 mM borate/NaOH buffer were investigated by electric conductivity and light transmittance measurements at 25 ºC. The parameters of the surfactant to polymer association processes such as the critical aggregation concentration and saturation of the polymer by surfactants were determined from plots of specific conductivity vs total surfactant concentration, [surfactant]tot = [SDoD] + [SDeC]. Through the results was not observed a specific link of polymer with the surfactant, implying therefore a phenomenon only cooperative association.

  13. Le premier Aurignacien en France méditerranéenne : un bilan

    Directory of Open Access Journals (Sweden)

    Frédéric Bazile

    2002-01-01

    Full Text Available Identifié au début des années 1970 (La Laouza puis l'Esquictio-Grapaou dans les Gorges du Gardon, l'Aurignacien initial, au sens d'antérieur à l'Aurignacien I classique (Archaïque? Aurignacien «0»? Protoaurignacien?, est également connu en Provence (Rainaude et dans le Bassin de l'Aude (grotte Tournai et sans doute Traouc de la Fado. Il était vraisemblablement présent à la Balauzière (sous un Aurignacien ancien classique et à la Grotte Nicolas (Gard à l'abri Rothschild et, plus au Nord, dans l'abri du Pécheur (Ardéche. Sa présence dans la moyenne vallée du Rhône à la grotte Mandrin (Drome est également vraisemblable, ouvrant un premier jalon (outre Roclaine nécessairement à revisiter vers des sites plus septentrionaux aux affinités typologiques et technologiques troublantes tels Arcy (Grotte du Renne et le Trou de la Mère Clochette. Ce "technocomplexe», bien situé sur le plan chronologique, témoignent d'une forte unité culturele de la Campante et la Vénétie à la Catalogne, en passant par la Ligurie, sans doute l'un des foyers pricipaux. Il apparaît brutalement à la fin de l'Interstade wûrmien, sus-jacent, avec ou sans lacune de sédimentation et/ou d'érosion, à des dépôts livrant des industries moustériennes de faciès très différents selon la région concernée.Identificado a inicios de 1970 en la garganta de Gardon, primero en La Laouza, después en Esquicho-Grapaou, el Aurihaciense inicial en sentido de anterioridad al Auriñaciense I clásico, (Arcaico, «0», Protoaurihaciense es también conocido en Provence (Rainaude y el en valle del Aude (Tournai y Traouc de la Fado. Esta también presente de manera clara en Balauzière (bajo un Auriñaciense antiguo clásico y en la Grotte Nicolas (Gard en el abri Rothschild y, más al norte, en el abri Pêcheur (Ardéche. Su presencia en el valle medio del Rôdano en la grotte Mandrin en Drome (otro yacimiento séria Roclaine sirve como jalon hacia sitios m

  14. Premier League academy soccer players' experiences of competing in a tournament bio-banded for biological maturation.

    Science.gov (United States)

    Cumming, Sean P; Brown, Daniel J; Mitchell, Siobhan; Bunce, James; Hunt, Dan; Hedges, Chris; Crane, Gregory; Gross, Aleks; Scott, Sam; Franklin, Ed; Breakspear, Dave; Dennison, Luke; White, Paul; Cain, Andrew; Eisenmann, Joey C; Malina, Robert M

    2017-06-19

    Individual differences in the growth and maturation have been shown to impact player performance and development in youth soccer. This study investigated Premier League academy players' experiences of participating in a tournament bio-banded for biological maturation. Players (N = 66) from four professional soccer clubs aged 11 and 14 years and between 85-90% of adult stature participated in a tournament. Players competed in three 11 vs 11 games on a full size pitch with 25-min halves. Sixteen players participated in four 15-min focus groups and were asked to describe their experiences of participating in the bio-banded tournament in comparison to age group competition. All players described their experience as positive and recommended the Premier League integrate bio-banding into the existing games programme. In comparison to age-group competitions, early maturing players described the bio-banded games more physically challenging, and found that they had to adapt their style of play placing a greater emphasis on technique and tactics. Late maturing players considered the games to be less physically challenging, yet appreciated the having more opportunity to use, develop and demonstrate their technical, physical, and psychological competencies. Bio-banding strategies appear to contribute positively towards the holistic development of young soccer players.

  15. Relationship between Game Location and Match Result with the Amount of Aggression: Iranian Premier League Football Teams

    Directory of Open Access Journals (Sweden)

    Farhad Alahvisi

    2015-12-01

    Full Text Available The purpose of this study was to investigate the relationship between game location (host advantage, match result (win, lose or tie and the level of aggression in football teams of the Iranian Premier League. The study population consisted of Premier League Football teams (League XIII, and 60 matches (related to 4 teams that were available for the researcher, were selected as the sample. The current study can be regarded as applied and descriptive, in terms of purpose and data collection, respectively. In order to collect data, the match videos of selected teams were studied, then the results of the observations were written and recorded using Roberts et al. (1999 aggression model. The results showed that no significant difference was found between teams' aggression in host and guest matches (p>0.05. Also, a significant difference was observed between aggression and match result and the behaviors were more in lost matches (p< 0.05. In fact, mental stress caused by the loss resulted to more aggression to win. Hence, a match result, physical aggression and players' position led to significant difference in players' aggression. Therefore, the control and management of aggressive behaviors, especially at the time of failure will result in improved performance and efficiency of football teams. Also, these behaviors can be minimized by providing necessary training on anger management and negative emotions control among players.

  16. The effect of match standard and referee experience on the objective and subjective match workload of English Premier League referees.

    Science.gov (United States)

    Weston, M; Bird, S; Helsen, W; Nevill, A; Castagna, C

    2006-06-01

    The aim of the present study was to examine the effect of match standard and referee experience on the objective and subjective workload of referees during English Premier League and Football League soccer matches. We also examined the relationship between heart rate (HR) and ratings of perceived exertion (RPE) for assessing match intensity in soccer referees. Heart rate responses were recorded using short-range telemetry and RPE scores were collected using a 10-point scale. Analysis revealed a significant relationship between mean match HR and match RPE scores (r=0.485, pReferee experience had no effect on match HR and RPE responses to Premier League and Football League matches. The results of the present study demonstrate the validity of using HR and RPE as a measure of global match intensity in soccer referees. Referee experience had no effect on the referees' objective and subjective match workload assessments, whereas match intensity was correlated to competition standard. These findings have implications for fitness preparation and evaluation in soccer referees. When progressing to a higher level of competition, referees should ensure that appropriate levels of fitness are developed in order to enable them to cope with an increase in physical match demands.

  17. Les premiers colons de l’ancienne Haïti et leurs attaches en métropole, à l’aube des premiers établissements (1650-1700)

    OpenAIRE

    Hroděj, Philippe

    2012-01-01

    Au moment où les premiers établissements durables voient le jour dans ce qui va devenir la partie française de Saint-Domingue, les colons ne sont qu’une poignée. Il est nécessaire d’abord de raisonner sur le nombre, source d’isolement, renforcé par le relief qui compartimente les différents quartiers, par l’éloignement des Petites Antilles, par des liens commerciaux longtemps aléatoires et par le fait dominant que sont les guerres quasi continues, du fait des délais d’application des traités ...

  18. 鸭出血性大肠杆菌O46分离株实验病理模型的建立%Establishment of an Experimental Pathological Model by Using EHEC O46 Isolate in Duck

    Institute of Scientific and Technical Information of China (English)

    于学辉; 李键; 程安春; 黄志宏; 罗薇; 冉丹丹

    2013-01-01

    [目的]为了建立鸭源出血性大肠杆菌(Enterohaemorrhagic E. coli, EHEC)O46分离株的实验病理模型并观察其在雏鸭体内的动态分布及组织病理学和超微病理学变化。[方法]本研究以 O46分离株通过口服、肌肉和皮下注射3种途径感染10日龄健康雏鸭,感染剂量均为0.5 mL/只(2×108CFU·mL-1),感染后2、4、6、12、24h剖杀、取样,以后每隔12h剖杀、取样并对剖解变化进行详细观察,在每个安排的时间点平行采集2只雏鸭的心、肝、脾、肺、肾、脑、食道、胸腺、十二指肠、空肠、盲肠、直肠、法氏囊、胰腺和气管等组织制备组织切片和超薄切片,通过H.E染色、醋酸铀、柠檬酸铅染色和免疫组化染色,对感染雏鸭的组织病理学变化及超微病理变化、细菌抗原定位进行观察。[结果]实验病理模型能复制出与自然感染相同的病例,除气管未检测到细菌抗原外,其余的组织均检测到细菌抗原,心、肺、脾、肾和肠道是感染的主要靶器官,抗原主要存在其感染细胞的细胞质中,阳性信号最早出现于心脏。尸体剖解、组织病理学及超微病理学观察表明,雏鸭人工感染鸭源EHEC O46分离株后主要病理学损害为浆膜广泛性纤维素性炎症,心、肝、脾、肺、肾、法氏囊、小肠、胰腺、脑等器官充血、出血、炎性细胞浸润,肾小管上皮细胞、肝细胞、心肌细胞、肠上皮细胞等实质细胞变性、坏死或凋亡,法氏囊淋巴细胞减少。超薄切片电镜观察可在心、肺、脾、肝和小肠中观察到大肠杆菌,其侵害的主要靶细胞包括淋巴细胞、巨噬细胞、肠道上皮细胞、心肌纤维细胞。[结论]鸭源EHEC O46分离株能使雏鸭发病和死亡,实验病理模型能复制出EHEC引起的出血性肠炎和溶血性尿毒症等病理变化,该分离株能在鸭体内进行大面积的侵嗜,心脏、

  19. Should Health Organizations Use Web 2.0 Media in Times of an Infectious Disease Crisis? An In-depth Qualitative Study of Citizens’ Information Behavior During an EHEC Outbreak

    Science.gov (United States)

    van Gemert-Pijnen, Julia E.W.C; Beaujean, Desirée J.M.A; Wentzel, Jobke; van Steenbergen, Jim E

    2012-01-01

    Background Web 2.0 media (eg, Facebook, Wikipedia) are considered very valuable for communicating with citizens in times of crisis. However, in the case of infectious disease outbreaks, their value has not been determined empirically. In order to be able to take full advantage of Web 2.0 media in such a situation, the link between these media, citizens’ information behavior, and citizens’ information needs has to be investigated. Objective The goal of our study was to assess citizens’ Web 2.0 media use during an infectious disease outbreak and to determine which Web 2.0 medium is used for which goal. With this information, we wanted to formulate recommendations for health organizations that consider using Web 2.0 media as part of their communication strategy during an infectious disease outbreak. Methods A total of 18 student participants kept an information diary for 4 weeks during the 2011 enterohemorrhagic E. coli (EHEC) outbreak in Germany. Of them, 9 lived at the epicenter of the outbreak and 9 of them at some distance. The diaries were supplemented by a qualitative pre-survey (demographics) and postsurvey (questioning their satisfaction with information provision during the outbreak). Results The Internet appeared to be the most popular medium for passively receiving EHEC-related information, with news websites and websites of newspapers as the most consulted sources. Twitter was used for receiving information to a small degree, while Facebook played virtually no role. Participants indicated that they thought information posted on Twitter or Facebook was not reliable or was out of place. When actively seeking information, online newspapers and wikis were important sources. Several causes for (dis)satisfaction with information provision were uncovered: source credibility, contradicting messages, and a need for closure. Conclusions During an infectious disease outbreak, our small sample of students did not see social media (like Facebook and Twitter) as

  20. REMOVAL OF CHEMICAL AND MICROBIAL CONTAMINANTS IN DRINKING WATER - WATTS PREMIER M-2400 POINT-OF-ENTRY REVERSE OSMOSIS DRINKINGWATER TREATMENT SYSTEM

    Science.gov (United States)

    The Watts Premier M-2400 POE RO Drinking Water Treatment System was tested at the NSF Drinking Water Treatment Systems Laboratory for removal of the viruses fr and MS2, the bacteria Brevundimonas diminuta, and chemicals aldicarb, benzene, cadmium, carbofuran, cesium, chl...

  1. ENVIRONMENTAL TECHNOLOGY VERIFICATION REPORT, REMOVAL OF ARSENIC IN DRINKING WATER: WATTS PREMIER M-SERIES M-15,000 REVERSE OSMOSIS TREATMENT SYSTEM

    Science.gov (United States)

    Verification testing of the Watts Premier M-Series M-15,000 RO Treatment System was conducted over a 31-day period from April 26, 2004, through May 26, 2004. This test was conducted at the Coachella Valley Water District (CVWD) Well 7802 in Thermal, California. The source water...

  2. ENVIRONMENTAL TECHNOLOGY VERIFICATION REPORT, REMOVAL OF ARSENIC IN DRINKING WATER: WATTS PREMIER M-SERIES M-15,000 REVERSE OSMOSIS TREATMENT SYSTEM

    Science.gov (United States)

    Verification testing of the Watts Premier M-Series M-15,000 RO Treatment System was conducted over a 31-day period from April 26, 2004, through May 26, 2004. This test was conducted at the Coachella Valley Water District (CVWD) Well 7802 in Thermal, California. The source water...

  3. REMOVAL OF CHEMICAL AND MICROBIAL CONTAMINANTS IN DRINKING WATER - WATTS PREMIER M-2400 POINT-OF-ENTRY REVERSE OSMOSIS DRINKINGWATER TREATMENT SYSTEM

    Science.gov (United States)

    The Watts Premier M-2400 POE RO Drinking Water Treatment System was tested at the NSF Drinking Water Treatment Systems Laboratory for removal of the viruses fr and MS2, the bacteria Brevundimonas diminuta, and chemicals aldicarb, benzene, cadmium, carbofuran, cesium, chl...

  4. “You are back home!”——On Premier Wen Jiabao’s Meeting with Japanese Orphans Delegation

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    <正>Beijing,November 11,2009 The sky was exceptionally blue after the first snow.The temperature was low,but people could feel the bright sunshine’s warmth.For the members of the Japanese orphans delegation, they felt especially so because they would soon meet Premier Wen Jiabao whom they had been longing to see.

  5. Les marqueurs d'aspect de dicto : 'à première vue', 'au premier abord', 'de prime abord'

    Directory of Open Access Journals (Sweden)

    Lenepveu Véronique

    2012-07-01

    Full Text Available On se propose d’étudier les locutions adverbiales "à première vue", "au premier abord", "de prime abord", trois locutions qui comportent une forme numérale ordinale et qui entretiennent des relations de proximité sémantique. Nous nous intéressons précisément au fonctionnement sémantique et pragmatique de ces locutions à valeur paradigmatisante (Nølke 1983, quand elles introduisent un point de vue (Anscombre & Ducrot 1983, Ducrot 1984, Nølke 1994. Dans cette perspective, nous faisons l’hypothèse d’un procès énonciatif, qui consiste à prendre en considération une situation, et qui vise à constituer un jugement stabilisé. Nous admettons alors que les locutions adverbiales 'à première vue', 'au premier abord', 'de prime abord', servent à sélectionner la phase initiale de ce procès énonciatif, ce qu’attestent les mises en corrélation possibles des trois marqueurs de point de vue avec des expressions qui signalent une évolution dans le temps du jugement du locuteur concernant une situation ('à mieux regarder', 'à y regarder de plus près', 'tout compte fait', 'tout bien considéré', .... L’étude de ces enchaînements, qui vont parfois au-delà du paragraphe, fait apparaître la capacité des trois locutions à initier un cadre (Charolles 1997, et à appeler un autre cadre dans lequel le jugement va être réévalué. En présentant le point de vue qu’elle introduit comme provisoire et en attente de confirmation, chacune de ces locutions anticipe ainsi sur la suite du texte, et s’inscrit dans une structure aspectuelle de dicto qui est celle d’un procès énonciatif de constitution d’un jugement stabilisé. Nous nous appuyons sur des énoncés attestés au XXème, sélectionnés sur "Frantext intégral", ou bien choisis dans Le Monde sur cederom (1995-96, 1999-2002.

  6. Glycemic index and glycemic load are associated with some cardiovascular risk factors among the PREMIER study participants

    Directory of Open Access Journals (Sweden)

    Pao-Hwa Lin

    2012-06-01

    Full Text Available Background: The clinical significance of glycemic index (GI and glycemic load (GL is inconclusive. Objective : This study was conducted to examine the association of GI and GL with clinical cardiovascular disease (CVD risk factors including body weight, blood pressure (BP, serum lipids, fasting glucose, insulin and homocysteine over time among the PREMIER participants. Design: PREMIER was an 18-month randomized lifestyle intervention trial, conducted from 2000 to 2002, designed to help participants reduce BP by following the Dietary Approaches to Stop Hypertension (DASH dietary pattern, losing weight, reducing sodium and increasing physical activity. GI and GL were estimated from 24 h diet recall data at baseline, 6 and 18 months after intervention. PROC MIXED model was used to examine the association of changes in GI or GL with changes in CVD risk factors. Results: A total of 756 randomized participants, 62% females and 34% African Americans and who averaged 50.0±0.3 years old and 95.3±0.7 kg, were included in this report. Neither GI nor GL changes was associated with changes in any risk factors at 6 months. At 18 months, however, the GI change was significantly and positively associated with total cholesterol (TC change only (p<0.05, β = 23.80±12.11 mg/dL or 0.62±0.31 mmol/L with a significant age interaction. The GL change was significantly associated with TC (p=0.02, β = 0.28±0.15 mg/dL or 0.01±0.00 mmol/L positively and with low density lipoprotein cholesterol (LDL-C changes negatively (p=0.03, β = − 0.01±0.00 mg/dL or −0.00±0.00 mmol/L, and significant age interactions were observed for both. Conclusion: GI and GL was associated with TC and LDL-C after controlling for energy, fat and fiber intake and other potential confounders and the associations were modified by age. Further investigation into this relationship is important because of its potential clinical impact.

  7. Surveillance of Musculoskeletal Symptoms and Anthropometric Variables among Four International Cricket Teams Competed in ACC Premier League Malaysia 2014

    Directory of Open Access Journals (Sweden)

    Srinivas Mondam, Rahul Shaik, Jalaj Jalaja Prakash, Jeffrey Low Fook, Sirisha Nekkanti

    2016-04-01

    Full Text Available Background and Purpose: Chronic musculoskeletal injuries are more common in cricket players. Acute problems may be due to trauma or injuries during sporting. The musculoskeletal system includes muscles, joints, bones, cartilage, ligaments, fascia, nerves and other associated soft tissues. Whatever the mode of injury, it causes pain, movement restriction, muscle weakness, and ultimately loss of functions. Anthropometric variables of each player in cricket will also influence the occurrence of problems. The current study focused on identifying the most common site involved in musculoskeletal problems and to explore possible variations in anthropometric characteristics. Methodology: This study was conducted in Kuala Lumpur, Malaysia where Asian Cricket Council Premier League 2014 was conducted. Permission to approach the players was taken from the council members and all the players were assured that the information collected from them will be kept confidential and all were explained about the objective study. Modified Nordic musculoskeletal questionnaire was distributed to the players and instructions were given about how to fill the questionnaire. Their anthropometric characteristics, experience and time of training sessions were collected by a blinded assessor. Results: Player's height (p = 0.003, weight (p = 0.050, experience (p = 0.001 and practicing hours per week (0.002 were analyzed. There is a statistically significant difference in these characteristics was observed. Occurrence of acute troubles (within 7 days of upper back and elbow region were found different in four teams with a P value of 0.007 and 0.022 respectively. Persistence of neck, shoulder and lower back troubles in the last one year has a significant difference between the groups with a P value of 0.014, 0.003 and 0.021 respectively. Conclusion: This study can conclude that the prevalence of musculoskeletal injuries is more in cricket. Especially shoulder, neck, lower

  8. Microfluidic Integration of a Cloth-Based Hybridization Array System (CHAS) for Rapid, Colorimetric Detection of Enterohemorrhagic Escherichia coli (EHEC) Using an Articulated, Centrifugal Platform.

    Science.gov (United States)

    Geissler, Matthias; Clime, Liviu; Hoa, Xuyen D; Morton, Keith J; Hébert, Harold; Poncelet, Lucas; Mounier, Maxence; Deschênes, Mylène; Gauthier, Martine E; Huszczynski, George; Corneau, Nathalie; Blais, Burton W; Veres, Teodor

    2015-10-20

    We describe the translation of a cloth-based hybridization array system (CHAS), a colorimetric DNA detection method that is used by food inspection laboratories for colony screening of pathogenic agents, onto a microfluidic chip format. We also introduce an articulated centrifugal platform with a novel fluid manipulation concept based on changes in the orientation of the chip with respect to the centrifugal force field to time the passage of multiple components required for the process. The platform features two movable and motorized carriers that can be reoriented on demand between 0 and 360° during stage rotation. Articulation of the chip can be used to trigger on-the-fly fluid dispensing through independently addressable siphon structures or to relocate solutions against the centrifugal force field, making them newly accessible for downstream transfer. With the microfluidic CHAS, we achieved significant reduction in the size of the cloth substrate as well as the volume of reagents and wash solutions. Both the chip design and the operational protocol were optimized to perform the entire process in a reliable, fully automated fashion. A demonstration with PCR-amplified genomic DNA confirms on-chip detection and identification of Escherichia coli O157:H7 from colony isolates in a colorimetric multiplex assay using rfbO157, fliCH7, vt1, and vt2 genes.

  9. Give me refuge : transforming the most contaminated square mile on earth into a premier urban wildlife refuge

    Energy Technology Data Exchange (ETDEWEB)

    Williams, L. [Environmental Protection Agency Region 8, Denver, CO (United States)

    2008-07-01

    The Rocky Mountain Arsenal (RMA) was a United States chemical weapons manufacturing center located in the Colorado, Denver area. This presentation described the transformation of this highly contaminated site into a premier urban wildlife refuge. It included background information on the RMA and described the manufacturing and disposal history, including munition storage, liquid waste disposal and an overview of groundwater contamination. Soil remedies were summarized with particular reference to the timeline of the reclamation; wildlife-remedy/refuge interaction; innovative technologies such as hex pits and a dioxin study; and nuisance odour projects. The RMA was described as a nationwide clean-up success as it was a winner of the 2007 revitalization award from the Environmental Protection Agency. The RMA was originally about 27 square miles in the greater Denver area, of which 1 square mile was transformed into the wildlife refuge. The Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) process was demonstrated in chart format. The presentation concluded with a discussion of lessons learned, such as sharing a mutual goal, and assuming that a mutually acceptable solution is possible. tabs., figs.

  10. Estimation de la survie des alevins de carpe (C. carpio au cours de leur premier mois d'existence

    Directory of Open Access Journals (Sweden)

    MOREAU J.

    1979-10-01

    Full Text Available Des observations réalisées sur deux stations piscicoles de Madagascar, dans des étangs ne recevant ni engrais ni nourriture, révèlent qu'au cours de leur premier mois les alevins de carpe (C. carpio subissent des mortalités voi-sines de 50 %. Ces dernières sont encore plus élevées en début et en fin de saison de reproduction. Au début, les fortes mortalités sont dues à la température trop basse et aux disponibilités alimentaires insuffisantes ; en fin de saison de reproduction, la température trop élevée et la mauvaise qualité des œufs sont sans doute en cause. Une fumure adéquate des étangs de grossissement des alevins et une alimentation correcte des géniteurs permettront peut-être de diminuer ces mortalités.

  11. Incidence, nature, and pattern of injuries to referees in a premier football (soccer) league: a prospective study.

    Science.gov (United States)

    Kordi, Ramin; Chitsaz, Alireza; Rostami, Mohsen; Mostafavi, Reza; Ghadimi, Mahmoodreza

    2013-09-01

    Despite the crucial role of referees in a soccer match, few researchers have targeted the injury profile of referees in their studies. Understanding the incidence, nature, and pattern of injuries could provide important information for educational and preventative efforts at the international level. The incidence rate and patterns of acute injuries to official referees of the Iranian Premier Football League during the 2009-2010 season are similar to those reported among referees in short-term international competitions such as FIFA World Cup. Prospective cohort study. Demographic data for 74 referees, including 30 main referees and 44 assistant referees, were collected at the beginning of the season. To record injuries and refereeing time, weekly contact was made by a physician. In total, 102 injuries were reported by referees during the football season. The incidence rates of injuries among referees during training and matches were 4.6 and 19.6 injuries per 1000 hours, respectively. Muscular and tendon injuries were found to be the most common type of injury, and the most common site of injury was the lower leg followed by the hip and groin. The results of this study are consistent with similar prospective studies evaluating injuries to referees over the course of a short-term tournament. These findings provide a base for suggesting possible preventive recommendations in future studies.

  12. Detection of Shiga toxin-producing Escherichia coli by sandwich enzyme-linked immunosorbent assay using chicken egg yolk IgY antibodies

    Directory of Open Access Journals (Sweden)

    Yanil R Parma

    2012-06-01

    Full Text Available Enterohemorrhagic Escherichia coli (EHEC, a subset of Shiga toxin producing E. coli (STEC is associated with a spectrum of diseases that includes diarrhea, hemorrhagic colitis and a life-threatening hemolytic uremic syndrome (HUS. Regardless of serotype, Shiga toxins (Stx1 and/or Stx2 are uniformly expressed by all EHEC, and so exploitable targets for laboratory diagnosis of these pathogens. In this study, a sandwich ELISA for determination of Shiga toxin (Stx was developed using anti-Stx2 B subunit antibodies and its performance was compared with that of the Vero cell assay and a commercial immunoassay kit. Chicken IgY was used as capture antibody and a HRP-conjugated rabbit IgG as the detection antibody. The anti-Stx2B IgY was harvested from eggs laid by hens immunized with a recombinant protein fragment. Several parameters were tested in order to optimize the sandwich ELISA assay, including concentration of antibodies, type and concentration of blocking agent, and incubation temperatures. Supernatants from 42 STEC strains of different serotypes and stx variants, including stx2EDL933, stx2vha, stx2vhb, stx2g, stx1EDL933 and stx1d were tested. All Stx variants were detected by the sandwich ELISA, with a detection limit of 400 ng /ml Stx2. Twenty three strains negative for stx genes, including different bacteria species, showed no activity in Vero cell assay and produced negative results in ELISA, except for 2 strains. Our results show that anti-Stx2B IgY sandwich ELISA could be used in routine diagnosis as a rapid, specific and economic method for detection of Shiga toxin-producing E. coli.

  13. Phonologie et morphosyntaxe de l’anglais dans un produit SIC : le premier module de MACAO Phonologie et morphosyntaxe de l’anglais dans un produit SIC : le premier module de MACAO

    Directory of Open Access Journals (Sweden)

    Laurence Vincent-Durroux

    2002-12-01

    Full Text Available La préoccupation fréquente, chez les étudiants spécialistes d’autres disciplines que l’anglais, d’améliorer leur compréhension de l’anglais oral a motivé notre démarche de création d’un produit SIC (Systèmes d’Information et de Communication : MACAO (Modules d’Aide à la Compréhension de l’Anglais Oral. Le premier module, "S’entraîner à la reconnaissance", est réalisé et l’outil informatique s’est révélé particulièrement utile pour notre projet. Dans cet article, nous faisons état des difficultés récurrentes en compréhension de l’anglais oral : elles peuvent être dues à des attentes erronées fondées sur la dissymétrie entre la langue écrite et la langue orale, mais aussi à une reconnaissance difficile de certains morphèmes par l’existence de variantes phonologiques en fonction du contexte et par la proximité phonologique de certains morphèmes. Nous présentons également comment le contenu du premier module tente d’apporter des solutions à ces difficultés : en amenant les apprenants à prendre conscience du phénomène de réduction vocalique et d’inaccentuation qui touche certaines syllabes des mots aussi bien que certains éléments monosyllabiques de l’énoncé et en proposant un entraînement à la reconnaissance de ces éléments. Le module est en cours de validation avec une phase d’évaluation.French students who have English as part of their curriculum often express the wish to improve their comprehension of oral English. This led us to envisage the creation of a CALL product: MACAO (Modules to help in the comprehension of oral English. The first module has been created: "Training oneself for recognition". The computer was particularly adequate in this prospect. In this paper we present the most frequent difficulties in the comprehension of oral English: they can be related either to erroneous expectations based on the dissymmetry between the written form and the oral

  14. Evidence-based hamstring injury prevention is not adopted by the majority of Champions League or Norwegian Premier League football teams: The Nordic Hamstring survey

    OpenAIRE

    Bahr, Roald; Thorborg, Kristian; Ekstrand, Jan

    2015-01-01

    Background: The Nordic hamstring (NH) exercise programme was introduced in 2001 and has been shown to reduce the risk of acute hamstring injuries in football by at least 50%. Despite this, the rate of hamstring injuries has not decreased over the past decade in male elite football. Aim: To examine the implementation of the NH exercise programme at the highest level of male football in Europe, the UEFA Champions League (UCL), and to compare this to the Norwegian Premier League, Tippeligaen...

  15. Case study: Muscle atrophy and hypertrophy in a premier league soccer player during rehabilitation from ACL injury.

    Science.gov (United States)

    Milsom, Jordan; Barreira, Paulo; Burgess, Darren J; Iqbal, Zafar; Morton, James P

    2014-10-01

    The onset of injury and subsequent period of immobilization and disuse present major challenges to maintenance of skeletal muscle mass and function. Although the characteristics of immobilization-induced muscle atrophy are well documented in laboratory studies, comparable data from elite athletes in free-living conditions are not readily available. We present a 6-month case-study account from a professional soccer player of the English Premier League characterizing rates of muscle atrophy and hypertrophy (as assessed by DXA) during immobilization and rehabilitation after ACL injury. During 8 weeks of inactivity and immobilization, where the athlete adhered to a low carbohydrate-high protein diet, total body mass decreased by 5 kg attributable to 5.8 kg loss and 0.8 kg gain in lean and fat mass, respectively. Changes in whole-body lean mass was attributable to comparable relative decreases in the trunk (12%, 3.8 kg) and immobilized limb (13%, 1.4 kg) whereas the nonimmobilized limb exhibited smaller declines (7%, 0.8 kg). In Weeks 8 to 24, the athlete adhered to a moderate carbohydrate-high protein diet combined with structured resistance and field based training for both the lower and upper-body that resulted in whole-body muscle hypertrophy (varying from 0.5 to 1 kg per week). Regional hypertrophy was particularly pronounced in the trunk and nonimmobilized limb during weeks 8 to 12 (2.6 kg) and 13 to 16 (1.3 kg), respectively, whereas the previously immobilized limb exhibited slower but progressive increases in lean mass from Week 12 to 24 (1.2 kg). The athlete presented after the totality of the injured period with an improved anthropometrical and physical profile.

  16. Continuations intra- et interphrastiques du français : premiers résultats expérimentaux

    Directory of Open Access Journals (Sweden)

    Bartkova Katarina

    2012-07-01

    Full Text Available Continuations intra et interphrastiques du français : premiers résultats expérimentaux Cet article rend compte d’un certain nombre d’observations pour l’étude des continuations mineures et majeures en français. Ces observations ont été obtenues dans le cadre d’un projet en cours sur les patrons prosodiques non-conclusifs en français et en anglais. Ici, Nous discutons plus particulièrement les variations de pente concernant deux types de configurations continuatives : (i le segment final d’un SN sujet dans une phrase simple déclarative, suivie ou non d’une autre phrase, (ii le segment final de X dans une suite complexe de type XY où X et Y sont des phrases simples reliées par une relation de discours, marquée ou non par une conjonction. A l’issu d’un protocole expérimental contrastant huit configurations proposées à trente deux sujets, nous avons relevé les valeurs de F0 toutes les 10 ms sur les segments finaux du SN et de X. En calculant le coefficient de la droite de régression correspondant à ce segment, nous avons contrasté les différentes configurations. Les résultats vont dans le sens de l’existence deux types de continuation, conformément à une intuition répandue dans la littérature, mais montrent aussi que leur différence doit se décrire à l’aide paramètres plus complexes que ce qui est généralement proposé.

  17. Adalimumab reduces hand bone loss in rheumatoid arthritis independent of clinical response: Subanalysis of the PREMIER study

    Directory of Open Access Journals (Sweden)

    Elden Aake

    2011-02-01

    Full Text Available Abstract Background Anti-TNF therapy has been shown to reduce radiographic joint damage in rheumatoid arthritis (RA independent of clinical response. This has previously not been examined for periarticular bone loss, the other characteristic feature of bone involvement in RA. The objective of this study was to examine if treatment with the TNF-α inhibitor adalimumab also could reduce periarticular bone loss in RA patients independent of disease activity. Methods RA patients were recruited from the PREMIER study and included 214 patients treated with methotrexate (MTX plus adalimumab and 188 patients treated with MTX monotherapy. Periarticular bone loss was assessed by digital X-ray radiogrammetry metacarpal cortical index (DXR-MCI. Change in DXR-MCI was evaluated in patients with different levels of clinical response, as assessed by changes in DAS28 score at 52 weeks and in mean C-reactive protein (CRP levels during follow-up. Results In the MTX group, there was a greater median DXR-MCI loss among patients with moderate and high disease activity compared to those in remission or with low disease activity (-3.3% vs. -2.2%, p = 0.01. In contrast, periarticular bone loss was independent of disease activity (-1.9% vs. -2.4%, p = 0.99 in the combination group. In the MTX group patients with a mean CRP of ≥ 10 mg/l lost significantly more DXR-MCI than patients with low CRP (-3.1% vs. -1.9%, p Conclusion Adalimumab in combination with MTX reduces periarticular bone loss independently of clinical response. These results support the hypothesis that TNF-α stimulates the osteoclast not only by the inflammatory pathway but do also have a direct effect on the osteoclast. Trial Registration ClinicalTrials (NCT: NCT001195663

  18. Le premier partenariat public-privé pour l’irrigation au Maroc : durable pour tous ?

    Directory of Open Access Journals (Sweden)

    Houdret Annabelle

    2016-03-01

    Full Text Available Les partenariats public-privé (PPP sont un phénomène relativement récent dans le secteur de l’irrigation ; le projet El Guerdane au Maroc est ainsi le premier de son genre. Inauguré en 2008, le projet alimente en eau 10 000 ha de plantations d’agrumes. Les banques internationales de développement le présentent comme un succès, mais l’impact sur le développement local est, au mieux, mitigé. Alors que certains agriculteurs ont bénéficié de cette initiative, d’autres ont été marginalisés, en termes d’accès à l’eau, aux terres fertiles et au développement. Fondé sur des recherches de terrain extensives conduites entre 2005 et 2013, l’article révèle trois problèmes cruciaux du projet PPP : des effets souvent négatifs sur les revenus des acteurs et sur le développement ; un partage inégal des coûts, des bénéfices et des risques entre les secteurs public et privé ; un impact environnemental incertain. Sur la base de ces résultats, l’étude situe le projet dans le contexte plus large de l’évolution des rapports de force politico-économiques au Maroc.

  19. Anti-hypertensive medicines prescribing for medical outpatients in a premier teaching hospital in Nigeria: a probable shift of paradigm.

    Science.gov (United States)

    Eshiet, Unyime I; Yusuff, Kazeem B

    2014-04-01

    Previous studies of anti-hypertensive medicines utilization pattern in Nigeria showed that Angiotensin converting enzyme inhibitors (ACEIs) were often the least prescribed. However, the appropriate use of ACEIs in the black population achieves good blood pressure control and provides additional long term cardio- and renovascular protection benefits. To assess the current utilization pattern of antihypertensive medicines with specific emphasis on identifying possible shift in the frequency of use of ACEIs. A prospective cross-sectional assessment of the current utilization pattern of anti-hypertensive medicines was conducted among 300 randomly selected cohort at a 900-bed premier Teaching Hospital located in Ibadan, Southwestern Nigeria. The current utilization pattern was compared with the results of a study conducted at the same site and published 10 years ago. Of the 300 random cohorts, a majority (79%) were females (237) with mean age 58.7 years (SD=2.81 years. Stage 2 hypertension was the most frequent diagnosis (54.3%). The utilization of ACEIs and long acting CCB (amlodipine) significantly increased from 8.6% and 21% (Ten years ago) to 29.93% and 36.68% respectively (p ACEIs were documented in 1.5% (3), while laboratory monitoring of serum potassium, urea and creatinine was conducted in only 37% (111) of cohort. Potentially harmful drug-drug interactions were identified in 25% (75) of cohorts, and the most frequent were ACEIs + NSAIDs (53.3%), ACEIs + amiloride / hydrochlorothiazide (22.6%). Anti-hypertensive medicines utilization has significantly shifted towards the increased use of ACEIs and long acting dihydropyridine CCBs. The use of thiazides and methyldopa has declined significantly. Physicians appeared more cognizant of the long term cardio- and renovascular benefits inherent in using ACEIs in a high cardiovascular risk group such as black hypertensive.

  20. Examining the External Training Load of an English Premier League Football Team With Special Reference to Acceleration.

    Science.gov (United States)

    Akenhead, Richard; Harley, Jamie A; Tweddle, Simon P

    2016-09-01

    Akenhead, R, Harley, J, and Tweddle, S. Examining the external training load of an English Premier League football team with special reference to acceleration. J Strength Cond Res 30(9): 2424-2432, 2016-Practitioners and coaches often use external training load variables such as distance run and the number of high-speed running (HSR) activities to quantify football training. However, an important component of the external load may be overlooked when acceleration activities are not considered. The aim of this study was to describe the within-microcycle distribution of external load, including acceleration, during in-season 1-game weeks in an elite football team. Global Positioning System technology was used to collect time-motion data from 12 representative 7-day microcycles across a competitive season (48 training days, 295 data sets). Training time, total distance (TD), high-speed running (HSR) distance (>5.8 m·s), sprint running distance (>6.7 m·s) and acceleration variables were recorded during each training session. Data were analysed for interday and interposition differences using mixed linear modeling. The distribution of external load was characterized by the second training day of the microcycle (5 days prematch) exhibiting the highest values for all variables of training load, with the fourth day (1 day prematch) exhibiting the lowest values. Central midfield players covered ∼8-16% greater TD than other positions excluding wide midfielders (p ≤ 0.03, d = 0.2-0.4) and covered ∼17% greater distance accelerating 1-2 m·s than central defenders (p = 0.03, d = 0.7). When expressed relative to training duration and TD, the magnitude of interday and interposition differences were markedly reduced (p = 0.03, d = 0.2-0.3). When managing the distribution of training load, practitioners should be aware of the intensity of training sessions and consider the density of external load within sessions.

  1. Microbead agglutination based assays

    KAUST Repository

    Kodzius, Rimantas

    2013-01-21

    We report a simple and rapid room temperature assay for point-of-care (POC) testing that is based on specific agglutination. Agglutination tests are based on aggregation of microbeads in the presence of a specific analyte thus enabling the macroscopic observation. Such tests are most often used to explore antibody-antigen reactions. Agglutination has been used for protein assays using a biotin/streptavidin system as well as a hybridization based assay. The agglutination systems are prone to selftermination of the linking analyte, prone to active site saturation and loss of agglomeration at high analyte concentrations. We investigated the molecular target/ligand interaction, explaining the common agglutination problems related to analyte self-termination, linkage of the analyte to the same bead instead of different microbeads. We classified the agglutination process into three kinds of assays: a two- component assay, a three-component assay and a stepped three- component assay. Although we compared these three kinds of assays for recognizing DNA and protein molecules, the assay can be used for virtually any molecule, including ions and metabolites. In total, the optimized assay permits detecting analytes with high sensitivity in a short time, 5 min, at room temperature. Such a system is appropriate for POC testing.

  2. Colorimetric protein assay techniques.

    Science.gov (United States)

    Sapan, C V; Lundblad, R L; Price, N C

    1999-04-01

    There has been an increase in the number of colorimetric assay techniques for the determination of protein concentration over the past 20 years. This has resulted in a perceived increase in sensitivity and accuracy with the advent of new techniques. The present review considers these advances with emphasis on the potential use of such technologies in the assay of biopharmaceuticals. The techniques reviewed include Coomassie Blue G-250 dye binding (the Bradford assay), the Lowry assay, the bicinchoninic acid assay and the biuret assay. It is shown that each assay has advantages and disadvantages relative to sensitivity, ease of performance, acceptance in the literature, accuracy and reproducibility/coefficient of variation/laboratory-to-laboratory variation. A comparison of the use of several assays with the same sample population is presented. It is suggested that the most critical issue in the use of a chromogenic protein assay for the characterization of a biopharmaceutical is the selection of a standard for the calibration of the assay; it is crucial that the standard be representative of the sample. If it is not possible to match the standard with the sample from the perspective of protein composition, then it is preferable to use an assay that is not sensitive to the composition of the protein such as a micro-Kjeldahl technique, quantitative amino acid analysis or the biuret assay. In a complex mixture it might be inappropriate to focus on a general method of protein determination and much more informative to use specific methods relating to the protein(s) of particular interest, using either specific assays or antibody-based methods. The key point is that whatever method is adopted as the 'gold standard' for a given protein, this method needs to be used routinely for calibration.

  3. Differentiation of ncRNAs from small mRNAs in Escherichia coli O157:H7 EDL933 (EHEC) by combined RNAseq and RIBOseq - ryhB encodes the regulatory RNA RyhB and a peptide, RyhP.

    Science.gov (United States)

    Neuhaus, Klaus; Landstorfer, Richard; Simon, Svenja; Schober, Steffen; Wright, Patrick R; Smith, Cameron; Backofen, Rolf; Wecko, Romy; Keim, Daniel A; Scherer, Siegfried

    2017-02-28

    While NGS allows rapid global detection of transcripts, it remains difficult to distinguish ncRNAs from short mRNAs. To detect potentially translated RNAs, we developed an improved protocol for bacterial ribosomal footprinting (RIBOseq). This allowed distinguishing ncRNA from mRNA in EHEC. A high ratio of ribosomal footprints per transcript (ribosomal coverage value, RCV) is expected to indicate a translated RNA, while a low RCV should point to a non-translated RNA. Based on their low RCV, 150 novel non-translated EHEC transcripts were identified as putative ncRNAs, representing both antisense and intergenic transcripts, 74 of which had expressed homologs in E. coli MG1655. Bioinformatics analysis predicted statistically significant target regulons for 15 of the intergenic transcripts; experimental analysis revealed 4-fold or higher differential expression of 46 novel ncRNA in different growth media. Out of 329 annotated EHEC ncRNAs, 52 showed an RCV similar to protein-coding genes, of those, 16 had RIBOseq patterns matching annotated genes in other enterobacteriaceae, and 11 seem to possess a Shine-Dalgarno sequence, suggesting that such ncRNAs may encode small proteins instead of being solely non-coding. To support that the RIBOseq signals are reflecting translation, we tested the ribosomal-footprint covered ORF of ryhB and found a phenotype for the encoded peptide in iron-limiting condition. Determination of the RCV is a useful approach for a rapid first-step differentiation between bacterial ncRNAs and small mRNAs. Further, many known ncRNAs may encode proteins as well.

  4. Absolute nuclear material assay

    Science.gov (United States)

    Prasad, Manoj K [Pleasanton, CA; Snyderman, Neal J [Berkeley, CA; Rowland, Mark S [Alamo, CA

    2012-05-15

    A method of absolute nuclear material assay of an unknown source comprising counting neutrons from the unknown source and providing an absolute nuclear material assay utilizing a model to optimally compare to the measured count distributions. In one embodiment, the step of providing an absolute nuclear material assay comprises utilizing a random sampling of analytically computed fission chain distributions to generate a continuous time-evolving sequence of event-counts by spreading the fission chain distribution in time.

  5. Anti-hypertensive medicines prescribing for medical outpatients in a premier teaching hospital in Nigeria: a probable shift of paradigm

    Directory of Open Access Journals (Sweden)

    Eshiet UI

    2014-06-01

    Full Text Available Background: Previous studies of anti-hypertensive medicines utilization pattern in Nigeria showed that Angiotensin converting enzyme inhibitors (ACEIs were often the least prescribed. However, the appropriate use of ACEIs in the black population achieves good blood pressure control and provides additional long term cardio- and renovascular protection benefits. Objective: To assess the current utilization pattern of anti-hypertensive medicines with specific emphasis on identifying possible shift in the frequency of use of ACEIs. Methods: A prospective cross-sectional assessment of the current utilization pattern of anti-hypertensive medicines was conducted among 300 randomly selected cohort at a 900-bed premier Teaching Hospital located in Ibadan, Southwestern Nigeria. The current utilization pattern was compared with the results of a study conducted at the same site and published 10 years ago. Results: Of the 300 random cohorts, a majority (79% were females (237 with mean age 58.7 years (SD=2.81 years. Stage 2 hypertension was the most frequent diagnosis (54.3%. The utilization of ACEIs and long acting CCB (amlodipine significantly increased from 8.6% and 21% (Ten years ago to 29.93% and 36.68% respectively (p ˂ 0.0001. The use of thiazide diuretic and methyldopa declined significantly from 39.4% and 23.3% (Ten years ago to 16.12% and 9.7% respectively (p ˂ 0.0001. Adverse drug reactions due to ACEIs were documented in 1.5% (3, while laboratory monitoring of serum potassium, urea and creatinine was conducted in only 37% (111 of cohort. Potentially harmful drug-drug interactions were identified in 25% (75 of cohorts, and the most frequent were ACEIs + NSAIDs (53.3%, ACEIs + amiloride / hydrochlorothiazide (22.6%. Conclusions: Anti-hypertensive medicines utilization has significantly shifted towards the increased use of ACEIs and long acting dihydropyridine CCBs. The use of thiazides and methyldopa has declined significantly. Physicians appeared

  6. Cell viability assays: introduction.

    Science.gov (United States)

    Stoddart, Martin J

    2011-01-01

    The measurement of cell viability plays a fundamental role in all forms of cell culture. Sometimes it is the main purpose of the experiment, such as in toxicity assays. Alternatively, cell viability can be used to -correlate cell behaviour to cell number, providing a more accurate picture of, for example, anabolic -activity. There are wide arrays of cell viability methods which range from the most routine trypan blue dye exclusion assay to highly complex analysis of individual cells, such as using RAMAN microscopy. The cost, speed, and complexity of equipment required will all play a role in determining the assay used. This chapter aims to provide an overview of many of the assays available today.

  7. Tube-Forming Assays.

    Science.gov (United States)

    Brown, Ryan M; Meah, Christopher J; Heath, Victoria L; Styles, Iain B; Bicknell, Roy

    2016-01-01

    Angiogenesis involves the generation of new blood vessels from the existing vasculature and is dependent on many growth factors and signaling events. In vivo angiogenesis is dynamic and complex, meaning assays are commonly utilized to explore specific targets for research into this area. Tube-forming assays offer an excellent overview of the molecular processes in angiogenesis. The Matrigel tube forming assay is a simple-to-implement but powerful tool for identifying biomolecules involved in angiogenesis. A detailed experimental protocol on the implementation of the assay is described in conjunction with an in-depth review of methods that can be applied to the analysis of the tube formation. In addition, an ImageJ plug-in is presented which allows automatic quantification of tube images reducing analysis times while removing user bias and subjectivity.

  8. Transgenic Animal Mutation Assays

    Institute of Scientific and Technical Information of China (English)

    Tao Chen; Ph.D.D.A.B.T.

    2005-01-01

    @@ The novel transgenic mouse and rat mutation assays have provided a tool for analyzing in vivo mutation in any tissue, thus permitting the direct comparison of cancer incidence with mutant frequency.

  9. Assays for thrombopoietin

    Energy Technology Data Exchange (ETDEWEB)

    McDonald, T.P.

    1977-01-01

    In summary, thrombopoietin levels have been determined indirectly by measuring thrombocytopoiesis in assay animals (platelet counting, measurement of isotope incorporation into newly formed platelets, changes in platelet sizes, or alterations in number and size of megakaryocytes) and by use of an immunoassay. Although much work remains, it seems clear at the present time that isotopic uptake into platelets of specially prepared assay mice (rebound-thrombocytosis) is superior to the other techniques now available for the measurement of thrombopoietin. However, the ideal assay for TSF which is specific, rapid, and inexpensive is yet to be developed. An immunoassay is in the development stage, but will require additional work before it can be utilized for the routine assay of TSF.

  10. Influence of Low Temperature Enzyme Maceration Techniques on Volatile Compounds of Semi-dry Wine Made with cv. Premier of Rabbiteye Blueberries (Vaccinium ashei

    Directory of Open Access Journals (Sweden)

    Xueling Gao

    2015-02-01

    Full Text Available A low temperature enzyme maceration treatment was conducted during fermentation process of semi-dry cv. Premier blueberry wine. The aim of this study was to investigate the influence of maceration conditions on the wine aroma. As a pre-treatment, blueberry must was divided into 6 samples which were respectively treated by pectinase with 6 different maceration conditions at 6°C 1day, 6°C 2 days, 6°C 3 days, 16°C 1 day, 16°C 2 days and 16°C 3 days. After that wines were obtained by fermentation with Saccharomyces cerevisiae. Volatile compounds of wines were analyzed by GC-MS. Overall, the typical aroma compounds of semi-dry cv. Premier wines were constituted by three groups of organic compounds including esters, alcohols and fatty acids. Isoamyl acetate, ethyl caprylate, ethyl decanoate, 2-phenethanol and 3-methylbutanoic acid, which occupied 60% of the typical volatile aromatic compounds, all had higher Odor Activity Values (OAVs in 6°C 3 days than other conditions. Maceration temperature and time had a significant effect on concentration and varieties of wines aroma substances. The results presented will help to better understand the aroma winemaking potential of this variety.

  11. New Rapid Spore Assay

    Science.gov (United States)

    Kminek, Gerhard; Conley, Catharine

    2012-07-01

    The presentation will detail approved Planetary Protection specifications for the Rapid Spore Assay for spacecraft components and subsystems. Outlined will be the research and studies on which the specifications were based. The research, funded by ESA and NASA/JPL, was conducted over a period of two years and was followed by limited cleanroom studies to assess the feasibility of this assay during spacecraft assembly.

  12. Against vaccine assay secrecy.

    Science.gov (United States)

    Herder, Matthew; Hatchette, Todd F; Halperin, Scott A; Langley, Joanne M

    2015-01-01

    Increasing the transparency of the evidence base behind health interventions such as pharmaceuticals, biologics, and medical devices, has become a major point of critique, conflict, and policy focus in recent years. Yet the lack of publicly available information regarding the immunogenicity assays upon which many important, widely used vaccines are based has received no attention to date. In this paper we draw attention to this critical public health problem by reporting on our efforts to secure vaccine assay information in respect of 10 vaccines through Canada's access to information law. We argue, under Canadian law, that the public health interest in having access to the methods for these laboratory procedures should override claims by vaccine manufacturers and regulators that this information is proprietary; and, we call upon several actors to take steps to ensure greater transparency with respect to vaccine assays, including regulators, private firms, researchers, research institutions, research funders, and journal editors.

  13. Against vaccine assay secrecy

    Science.gov (United States)

    Herder, Matthew; Hatchette, Todd F; Halperin, Scott A; Langley, Joanne M

    2015-01-01

    Increasing the transparency of the evidence base behind health interventions such as pharmaceuticals, biologics, and medical devices, has become a major point of critique, conflict, and policy focus in recent years. Yet the lack of publicly available information regarding the immunogenicity assays upon which many important, widely used vaccines are based has received no attention to date. In this paper we draw attention to this critical public health problem by reporting on our efforts to secure vaccine assay information in respect of 10 vaccines through Canada's access to information law. We argue, under Canadian law, that the public health interest in having access to the methods for these laboratory procedures should override claims by vaccine manufacturers and regulators that this information is proprietary; and, we call upon several actors to take steps to ensure greater transparency with respect to vaccine assays, including regulators, private firms, researchers, research institutions, research funders, and journal editors. PMID:25826194

  14. Rover waste assay system

    Energy Technology Data Exchange (ETDEWEB)

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

    1997-11-01

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

  15. CTL ELISPOT assay.

    Science.gov (United States)

    Ranieri, Elena; Popescu, Iulia; Gigante, Margherita

    2014-01-01

    Enzyme-linked immune absorbent spot (Elispot) is a quantitative method for measuring relevant parameters of T cell activation. The sensitivity of Elispot allows the detection of low-frequency antigen-specific T cells that secrete cytokines and effector molecules, such as granzyme B and perforin. Cytotoxic T cell (CTL) studies have taken advantage with this high-throughput technology by providing insights into quantity and immune kinetics. Accuracy, sensitivity, reproducibility, and robustness of Elispot resulted in a wide range of applications in research as well as in diagnostic field. Actually, CTL monitoring by Elispot is a gold standard for the evaluation of antigen-specific T cell immunity in clinical trials and vaccine candidates where the ability to detect rare antigen-specific T cells is of relevance for immune diagnostic. The most utilized Elispot assay is the interferon-gamma (IFN-γ) test, a marker for CD8(+) CTL activation, but Elispot can also be used to distinguish different subsets of activated T cells by using other cytokines such as T-helper (Th) 1-type cells (characterized by the production of IFN-γ, IL-2, IL-6, IL-12, IL-21, and TNF-α), Th2 (producing cytokines like IL-4, IL-5, IL-10, and IL-13), and Th17 (IL-17) cells. The reliability of Elispot-generated data, by the evaluation of T cell frequency recognizing individual antigen/peptide, is the core of this method currently applied widely to investigate specific immune responses in cancer, infections, allergies, and autoimmune diseases. The Elispot assay is competing with other methods measuring single-cell cytokine production, e.g., intracellular cytokine by FACS or Miltenyi cytokine secretion assay. Other types of lymphocyte frequency and function assays include limiting dilution assay (LDA), cytotoxic T cell assay (CTL), and tetramer staining. Compared with respect to sensitivity the Elispot assay is outranking other methods to define frequency of antigen-specific lymphocytes. The method

  16. Assays for calcitonin receptors

    Energy Technology Data Exchange (ETDEWEB)

    Teitelbaum, A.P.; Nissenson, R.A.; Arnaud, C.D.

    1985-01-01

    The assays for calcitonin receptors described focus on their use in the study of the well-established target organs for calcitonin, bone and kidney. The radioligand used in virtually all calcitonin binding studies is /sup 125/I-labelled salmon calcitonin. The lack of methionine residues in this peptide permits the use of chloramine-T for the iodination reaction. Binding assays are described for intact bone, skeletal plasma membranes, renal plasma membranes, and primary kidney cell cultures of rats. Studies on calcitonin metabolism in laboratory animals and regulation of calcitonin receptors are reviewed.

  17. New oligosaccharyltransferase assay method.

    Science.gov (United States)

    Kohda, Daisuke; Yamada, Masaki; Igura, Mayumi; Kamishikiryo, Jun; Maenaka, Katsumi

    2007-11-01

    We developed a new in vitro assay for oligosaccharyltransferase (OST), which catalyzes the transfer of preassembled oligosaccharides on lipid carriers onto asparagine residues in polypeptide chains. The asparagine residues reside in the sequon, Asn-X-Thr/Ser, where X can be any amino acid residue except Pro. We demonstrate the potency of our assay using the OST from yeast. In our method, polyacrylamide gel electrophoresis is used to separate the glycopeptide products from the peptide substrates. The substrate peptide is fluorescently labeled and the formation of glycopeptides is analyzed by fluorescence gel imaging. Two in vitro OST assay methods are now widely used, but both the methods depend on previous knowledge of the oligosaccharide moiety: One method uses lectin binding as the separation mechanism and the other method uses biosynthetically or chemoenzymatically synthesized lipid-linked oligosaccharides as donors. N-linked protein glycosylation is found in all three domains of life, but little is known about the N-glycosylation in Archaea. Thus, our new assay, which does not require a priori knowledge of the oligosaccharides, will be useful in such cases. Indeed, we have detected the OST activity in the membrane fraction from a hyperthermophilic archaeon, Pyrococcus furiosus.

  18. Hyaluronic Acid Assays

    DEFF Research Database (Denmark)

    Itenov, Theis S; Kirkby, Nikolai S; Bestle, Morten H

    2015-01-01

    BACKGROUD: Hyaluronic acid (HA) is proposed as a marker of functional liver capacity. The aim of the present study was to compare a new turbidimetric assay for measuring HA with the current standard method. METHODS: HA was measured by a particle-enhanced turbidimetric immunoassay (PETIA) and enzyme...

  19. Instrument for assaying radiation

    Energy Technology Data Exchange (ETDEWEB)

    Coleman, Jody Rustyn; Farfan, Eduardo B.

    2016-03-22

    An instrument for assaying radiation includes a flat panel detector having a first side opposed to a second side. A collimated aperture covers at least a portion of the first side of the flat panel detector. At least one of a display screen or a radiation shield may cover at least a portion of the second side of the flat panel detector.

  20. Technical and tactical training team «Helios» Kharkiv in the first round of 23 Ukrainian football championship in the premier league 2013–2014

    Directory of Open Access Journals (Sweden)

    Rebaz Sleman

    2014-10-01

    Full Text Available Purpose: to define the characteristics of the model command of technical and tactical training team participating in the Ukrainian Premier League First League. Material and Methods: the research was conducted using the method of peer review. The experts were involved 5 specialists football. Results: the mean values for the analyzed variables in 10 games. The various technical and tactical actions and their percentage in the overall structure of the game team statistics for 20 games, as well as some indicators of team play "Helios" Kharkov. Conclusions: the obtained quantitative and qualitative indicators (coefficient of marriage as a team on the technical and tactical actions, as well as separately for each technical and tactical reception. The performances allow you to make adjustments to the training process this command to improve sportsmanship.

  1. 用Premiere软件为MTV视频添加卡拉OK字幕的方法%Methods to add Karaoke caption for MTV video using software Premiere

    Institute of Scientific and Technical Information of China (English)

    张如

    2012-01-01

    提出了在Premiere中为MTV视频添加卡拉OK字幕的两种制作途径。一是直接在Premiere中制作卡拉OK字幕,在Premiere中制作又可分为重叠字幕做法、轨道蒙版键做法、彩色蒙版做法三种方法。二是结合制作卡拉OK字幕的第三方插件来制作。%There are two approaches in adding Karaoke caption for MTV video using software Premiere. One is directly to do karaoke caption in the software, withal which includes superposition caption, typewriting by keyboard on the orbital masks, and color masks; another one is that Karaoke caption is added by third party plug-in which has an ability to do Karaoke caption.

  2. Données récentes sur les habitats ruraux du premier âge du Fer en Centre-Ouest

    OpenAIRE

    Maitay, Christophe

    2016-01-01

    Cette contribution propose de faire le point sur les modalités d’occupation des campagnes du centre-ouest de la France entre le IXe et le IVe s. av. J.‑C. Le développement de l’archéologie préventive permet depuis quelques années de préciser la question de l’occupation des campagnes au cours de la première moitié du premier millénaire avant notre ère. La multiplication des opérations et l’opportunité d’intervenir sur de grandes surfaces favorisent aujourd’hui les possibilités de saisir non se...

  3. Quantification of training load during one-, two- and three-game week schedules in professional soccer players from the English Premier League: implications for carbohydrate periodisation.

    Science.gov (United States)

    Anderson, Liam; Orme, Patrick; Di Michele, Rocco; Close, Graeme L; Morgans, Ryland; Drust, Barry; Morton, James P

    2016-01-01

    Muscle glycogen is the predominant energy source for soccer match play, though its importance for soccer training (where lower loads are observed) is not well known. In an attempt to better inform carbohydrate (CHO) guidelines, we quantified training load in English Premier League soccer players (n = 12) during a one-, two- and three-game week schedule (weekly training frequency was four, four and two, respectively). In a one-game week, training load was progressively reduced (P 14.4 km · h(-1) (14%, 18% and 23% in the one-, two- and three-game weeks, respectively). Considering that high CHO availability improves physical match performance but high CHO availability attenuates molecular pathways regulating training adaptation (especially considering the low daily customary loads reported here, e.g., 3-5 km per day), we suggest daily CHO intake should be periodised according to weekly training and match schedules.

  4. See you at the match: Motivation for sport consumption and intrinsic psychological reward of premier football league spectators in South Africa

    Directory of Open Access Journals (Sweden)

    Frederick W. Stander

    2016-04-01

    Full Text Available Orientation: Local football contributes significantly to the social- and economic welfare of South Africa through its spectators. Understanding the motives and experiences of football spectators could provide opportunities for capitalising on football as revenue stream feeding the South African economy. Research purpose: To investigate how motives for sport consumption predict intrinsic psychological reward of South African premier league football spectators. Motivation for the study: Sport - particularly football - is an untapped resource for stimulating economic development and growth through its consumers. Spectators, who often experience their investment in the sport as deeply rewarding and meaningful, should participate more frequently in purchasing products or services associated with the sport. Through understanding the motives for sport consumption of South African premier league football spectators and the impact of these motives on intrinsic psychological reward experiences, football clubs are able to provide a targeted experience or service to spectators in order to further stimulate economic growth. Research design, approach and method: A census sample of 806 football spectators attending various matches at a football stadium in Soweto was drawn. A cross-sectional research design was implemented. This research was exploratory and descriptive. Structural equation modelling was implemented to assess the factor structures of the constructs, to confirm composite reliability of the measures and to assess the structural paths between the variables. Main findings: A predictive model for intrinsic psychological rewards (life satisfaction and meaning through the motivation for sport consumption (individual – and game related factors was confirmed. It was further established that motivation for sport consumption is significantly positively a related to and b associated with the experience of intrinsic psychological reward by South African

  5. The corneal pocket assay.

    Science.gov (United States)

    Ziche, Marina; Morbidelli, Lucia

    2015-01-01

    The cornea in most species is physiologically avascular, and thus this assay allows the measurement of newly formed vessels. The continuous monitoring of neovascular growth in the same animal allows the evaluation of drugs acting as suppressors or stimulators of angiogenesis. Under anesthesia a micropocket is produced in the cornea thickness and the angiogenesis stimulus (tumor tissue, cell suspension, growth factor) is placed into the pocket in order to induce vascular outgrowth from the limbal capillaries. Neovascular development and progression can be modified by the presence of locally released or applied inhibitory factors or by systemic treatments. In this chapter the experimental details of the avascular cornea assay, the technical challenges, and advantages and disadvantages in different species are discussed. Protocols for local drug treatment and tissue sampling for histology and pharmacokinetic profile are reported.

  6. Kinetic Tetrazolium Microtiter Assay

    Science.gov (United States)

    Pierson, Duane L.; Stowe, Raymond; Koenig, David

    1993-01-01

    Kinetic tetrazolium microtiter assay (KTMA) involves use of tetrazolium salts and Triton X-100 (or equivalent), nontoxic, in vitro color developer solubilizing colored metabolite formazan without injuring or killing metabolizing cells. Provides for continuous measurement of metabolism and makes possible to determine rate of action of antimicrobial agent in real time as well as determines effective inhibitory concentrations. Used to monitor growth after addition of stimulatory compounds. Provides for kinetic determination of efficacy of biocide, greatly increasing reliability and precision of results. Also used to determine relative effectiveness of antimicrobial agent as function of time. Capability of generating results on day of test extremely important in treatment of water and waste, disinfection of hospital rooms, and in pharmaceutical, agricultural, and food-processing industries. Assay also used in many aspects of cell biology.

  7. B cell helper assays.

    Science.gov (United States)

    Abrignani, Sergio; Tonti, Elena; Casorati, Giulia; Dellabona, Paolo

    2009-01-01

    Activation, proliferation and differentiation of naïve B lymphocytes into memory B cells and plasma cells requires engagement of the B cell receptor (BCR) coupled to T-cell help (1, 2). T cells deliver help in cognate fashion when they are activated upon recognition of specific MHC-peptide complexes presented by B cells. T cells can also deliver help in a non-cognate or bystander fashion, when they do not find specific MHC-peptide complexes on B cells and are activated by alternative mechanisms. T-cell dependent activation of B cells can be studied in vitro by experimental models called "B cell helper assays" that are based on the co-culture of B cells with activated T cells. These assays allow to decipher the molecular bases for productive T-dependent B cell responses. We show here examples of B cell helper assays in vitro, which can be reproduced with any subset of T lymphocytes that displays the appropriate helper signals.

  8. Growth cone collapse assay.

    Science.gov (United States)

    Cook, Geoffrey M W; Jareonsettasin, Prem; Keynes, Roger J

    2014-01-01

    The growth cone collapse assay has proved invaluable in detecting and purifying axonal repellents. Glycoproteins/proteins present in detergent extracts of biological tissues are incorporated into liposomes, added to growth cones in culture and changes in morphology are then assessed. Alternatively purified or recombinant molecules in aqueous solution may be added directly to the cultures. In both cases after a defined period of time (up to 1 h), the cultures are fixed and then assessed by inverted phase contrast microscopy for the percentage of growth cones showing a collapsed profile with loss of flattened morphology, filopodia, and lamellipodia.

  9. FLUIDICS DEVICE FOR ASSAY

    DEFF Research Database (Denmark)

    2007-01-01

    The present invention relates to a device for use in performing assays on standard laboratory solid supports whereon chemical entities are attached. The invention furthermore relates to the use of such a device and a kit comprising such a device. The device according to the present invention is a......, when operatively connected, one or more chambers (21) comprising the chemical entities (41), the inlet(s) (5) and outlet(s) (6) and chambers (21) being in fluid connection. The device further comprise means for providing differing chemical conditions in each chamber (21)....

  10. Radon assay for SNO+

    Energy Technology Data Exchange (ETDEWEB)

    Rumleskie, Janet [Laurentian University, Greater Sudbury, Ontario (Canada)

    2015-12-31

    The SNO+ experiment will study neutrinos while located 6,800 feet below the surface of the earth at SNOLAB. Though shielded from surface backgrounds, emanation of radon radioisotopes from the surrounding rock leads to back-grounds. The characteristic decay of radon and its daughters allows for an alpha detection technique to count the amount of Rn-222 atoms collected. Traps can collect Rn-222 from various positions and materials, including an assay skid that will collect Rn-222 from the organic liquid scintillator used to detect interactions within SNO+.

  11. RAS - Screens & Assays - Drug Discovery

    Science.gov (United States)

    The RAS Drug Discovery group aims to develop assays that will reveal aspects of RAS biology upon which cancer cells depend. Successful assay formats are made available for high-throughput screening programs to yield potentially effective drug compounds.

  12. Bacterial assays for recombinagens.

    Science.gov (United States)

    Hoffmann, G R

    1992-12-01

    Two principal strategies have been used for studying recombinagenic effects of chemicals and radiation in bacteria: (1) measurement of homologous recombination involving defined alleles in a partially diploid strain, and (2) measurement of the formation and loss of genetic duplications in the bacterial chromosome. In the former category, most methods involve one allele in the bacterial chromosome and another in a plasmid, but it is also possible to detect recombination between two chromosomal alleles or between two extrachromosomal alleles. This review summarizes methods that use each of these approaches for detecting recombination and tabulates data on agents that have been found to be recombinagenic in bacteria. The assays are discussed with respect to their effectiveness in testing for recombinagens and their potential for elucidating mechanisms underlying recombinagenic effects.

  13. Les premiers romans noirs français : simples exercices de style ou trahisons littéraires complexes ?

    Directory of Open Access Journals (Sweden)

    Anne Cadin

    2011-02-01

    Full Text Available Les premiers romans noirs français, imités des hard-boiled novels, ont été écrits sous pseudonyme : Malet devient Harding ; Vian, Sullivan ; Queneau, Mara et Arcouët, Stewart. Quelle part de trahison anime ces auteurs ? Sert-elle à faire vendre ou à dénoncer l’invasion de la culture américaine et le goût du scandale ? Si stratégie publicitaire et séduction du lecteur entrent en jeu, le processus de trahison est néanmoins volontairement miné par ces romanciers : il devient source de réflexion sur la lecture et l’écriture.The first French romans noirs, modeled on the so-called “hard-boiled” novels, were written under pseudonyms : Malet becomes Harding ; Vian, Sullivan ; Queneau, Mara, and Arcouët, Stewart. To what degree are these authors motivated by betrayal ? Is it employed merely for marketing purposes, or to denounce the invasion of American culture and a certain taste for scandal ? If advertising tactics and the seduction of the reader are undeniable elements of such novels, these writers nevertheless consciously sabotage the process of betrayal, transforming it into a source of reflection about reading and writing.

  14. Management of chronic recurrent osteitis pubis/pubic bone stress in a Premier League footballer: Evaluating the evidence base and application of a nine-point management strategy.

    Science.gov (United States)

    McAleer, Stephen S; Gille, Justus; Bark, Stefan; Riepenhof, Helge

    2015-08-01

    The aim of this paper was to use a clinical example to describe a treatment strategy for the management of recurrent chronic groin pain and evaluate the evidence of the interventions. A professional footballer presented with chronic recurrent OP/PBS. The injury was managed successfully with a nine-point programme - 1. Acute pharmacological management. 2. Tone reduction of over-active structures. 3. Improved ROM at hips, pelvis and thorax. 4. Adductor strength. 5. Functional movement assessment. 6. Core stability. 7. Lumbo-pelvic control. 8. Gym-based strengthening. 9. Field-based conditioning/rehabilitation. The evidence for these interventions is reviewed. The player returned to full training and match play within 41 and 50 days, respectively, and experienced no recurrence of his symptoms in follow up at 13 months. This case report displays a nine-point conservative management strategy for OP/PBS, with non-time dependent clinical objective markers as the progression criteria in a Premier League football player. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Aux sources de l’herméneutique occidentale :les premiers commentaires dans les traditions grecque, juive et chrétienne

    Directory of Open Access Journals (Sweden)

    Christophe Rico

    2003-09-01

    Full Text Available L’article nuance, à propos de l’Antiquité, la distinction établie par Michel Charles entre rhétorique, entendue au sens de « production littéraire », et commentaire : dans le premier cas une œuvre littéraire constitue un modèle d’écriture que l’on peut imiter et parfaire ; dans le second, elle est transmise comme un monument intouchable que l’on peut citer et commenter, mais non pas imiter.Dans les traditions grecque, juive et chrétienne, le commentaire, en effet, doit plutôt être considéré comme une œuvre littéraire au second degré dans la mesure où il se fonde sur un texte commenté qui l’engendre et le suscite. L’article évoque dans cette perspective l’herméneutique du mot dans l’école d’Alexandrie, l’herméneutique de l’énoncé dans les écoles de philosophie antiques, les commentaires juifs anciens et médiévaux, l’herméneutique chrétienne ancienne, et leurs variations autour du sens littéral et du sens spirituel.

  16. The Effect of Autonomy-Supportive Behaviors of Coaches on Need Satisfaction and Sport Commitment of Elite Female Players in Handball Premier League

    Directory of Open Access Journals (Sweden)

    Sedigheh Hosseinpoor Delavar

    2012-01-01

    Full Text Available This study determined the effects of autonomy-supportive behaviors on satisfying the psychological needs and commitment of female handball players in premier league in Iran. Here, we used descriptive research (survey method. The statistical population of 237 players was selected as our samples. We administered three questionnaires for autonomy-Supportive behaviors, psychological needs and commitment including perceived autonomy-supportive behaviors of coaches in sport (PASSES, satisfaction of the psychological needs in sports and sports Commitment Scale (SCMS questionnaires. We applied multiple regression analysis and structural equation modeling (SEM to analyze the data. Results showed there was a positive correlation between autonomy-supportive behaviors with the psychological needs of competence and commitment of athletes. On the other hand, the players `commitment correlated positively with psychological needs. Results of multivariate regression showed that the autonomy-supportive behaviors were predictor of psychological needs and commitment of players. The path analysis, also, established mediator role of psychological needs between autonomy-supportive behavior of the coach and players` commitment. Thus, the self-determination Theory among the elite players and sports teams was confirmed.

  17. Herbicide resistance screening assay.

    Science.gov (United States)

    Peterson, Joan M

    2009-01-01

    Herbicide resistance screening is a method that can be used not only to determine presence of the enzyme, phosphinothricin acetyltransferase, encoded by either the Bar or the Pat gene in transgenic maize, but also to assess the inheritance ratio of those genes in a segregating population. Herbicide screening can also be used to study linkage of a transgene of interest that was cotransformed with the herbicide resistance marker gene. By combining the herbicide screen assay with a PCR-based screen of leaf tissue DNA for the presence of both the Bar or the Pat gene marker and a cotransformed transgene of interest from the same seedling tissue and maintaining that seedling identity, the researcher can identify linkage or the possible breakdown in linkage of the marker gene and the transgene of interest. Further, the occurrence of "DNA silencing" can be evaluated if an individual seedling that was susceptible to the applied herbicide nonetheless gave PCR data that indicated presence of the gene responsible for herbicide resistance. Similarly, "DNA silencing" of the gene of interest may be investigated if the seeds can be screened and scored for that phenotypic trait in a nondestructive manner prior to planting.

  18. Quantitative Detection of Shiga Toxins Directly from Stool Specimens of Patients Associated with an Outbreak of Enterohemorrhagic Escherichia coli in Japan—Quantitative Shiga toxin detection from stool during EHEC outbreak

    Directory of Open Access Journals (Sweden)

    Eiki Yamasaki

    2015-10-01

    Full Text Available Detection of Shiga toxins (Stx is important for accurate diagnosis of Enterohemorrhagic Escherichia coli infection. In this study, we quantitatively analyzed Stx protein in nine patients’ stool during an outbreak that occurred in Japan. Highly sensitive immunoassay (bead enzyme-linked immunosorbent assay (bead-ELISA revealed that the concentrations of toxins in stool of patients ranged from 0.71 to 10.44 ng/mL for Stx1 and 2.75 to 51.61 ng/mL for Stx2. To our knowledge, this is the first report that reveals the range of Stx protein concentrations in human stools.

  19. Quantitative Detection of Shiga Toxins Directly from Stool Specimens of Patients Associated with an Outbreak of Enterohemorrhagic Escherichia coli in Japan—Quantitative Shiga toxin detection from stool during EHEC outbreak

    Science.gov (United States)

    Yamasaki, Eiki; Watahiki, Masanori; Isobe, Junko; Sata, Tetsutaro; Nair, G. Balakrish; Kurazono, Hisao

    2015-01-01

    Detection of Shiga toxins (Stx) is important for accurate diagnosis of Enterohemorrhagic Escherichia coli infection. In this study, we quantitatively analyzed Stx protein in nine patients’ stool during an outbreak that occurred in Japan. Highly sensitive immunoassay (bead enzyme-linked immunosorbent assay (bead-ELISA)) revealed that the concentrations of toxins in stool of patients ranged from 0.71 to 10.44 ng/mL for Stx1 and 2.75 to 51.61 ng/mL for Stx2. To our knowledge, this is the first report that reveals the range of Stx protein concentrations in human stools. PMID:26516915

  20. Perceptions of the Coach–Athlete Relationship Predict the Attainment of Mastery Achievement Goals Six Months Later: A Two-Wave Longitudinal Study among F. A. Premier League Academy Soccer Players

    Directory of Open Access Journals (Sweden)

    Adam R. Nicholls

    2017-05-01

    Full Text Available All football teams that compete within the F. A. Premier League possess an academy, whose objective is to produce more and better home-grown players that are capable of playing professionally. These young players spend a large amount of time with their coach, but little is known about player’s perception of the coach–athlete relationship within F. A. Premier League Academies. The objectives of this study were to examine whether perceptions of the coach–athlete relationship changed over six months and if the coach–athlete relationship predicted self-reported goal achievement among F. A. Premier League academy players. This study included cross-sectional (n = 104 and longitudinal (n = 52 assessments, in which academy soccer players completed a measure of the coach–athlete relationship and goal achievement across either one or two time periods. The cross-sectional data were subjected to bivariate correlations, whereas the longitudinal data were analyzed using multiple regressions. Perceptions of the coach–athlete relationship remained stable over time. The coach–athlete relationship predicted the achievement of mastery goals six months later. Enhancing the quality of the coach–athlete relationship among elite adolescent athletes appears to be a suitable way of maximizing mastery achievement goals, particularly among developmental athletes who participate in team sports.

  1. Disagreement between Human Papillomavirus Assays

    DEFF Research Database (Denmark)

    Rebolj, Matejka; Preisler, Sarah; Ejegod, Ditte Møller

    2014-01-01

    assays. Positive agreement between the assays was measured as the conditional probability that the results of all compared assays were positive given that at least one assay returned a positive result. Of all 5,064 samples, 1,679 (33.2%) tested positive on at least one of the assays. Among these, 41......We aimed to determine the disagreement in primary cervical screening between four human papillomavirus assays: Hybrid Capture 2, cobas, CLART, and APTIMA. Material from 5,064 SurePath samples of women participating in routine cervical screening in Copenhagen, Denmark, was tested with the four......% tested positive on all four. Agreement was lower in women aged ≥ 30 years (30%, vs. 49% at samples (29%, vs. 38% in follow-up samples), and in women with concurrent normal cytology (22%, vs. 68% with abnormal cytology). Among primary screening samples from women aged 30...

  2. L’Islam des pierres : l’expression de la foi dans les graffiti arabes des premiers siècles

    Directory of Open Access Journals (Sweden)

    Frédéric Imbert

    2014-07-01

    Full Text Available Les graffiti arabes coufiques des premiers siècles de l’islam, en Arabie comme au Proche-Orient, représentent une source d’information inépuisable sur la société musulmane des origines. Toutefois, la datation des textes du ier/viie siècle, antérieurs aux Umayyades, reste problématique et doit se fonder sur des analyses paléographiques rigoureuses. L’étude du contenu des graffiti relatifs à la foi peut aider à dater ces textes du fait qu’ils connurent des phases progressives de développement. Les plus anciens graffiti datés de 23/643 et 24/644 ne contiennent pas de référence au religieux ; les auteurs, comme leurs contemporains, semblèrent plus intéressés de pérenniser leurs noms sur la pierre, s’inscrivant dans la tradition safaïtique. La question des premières professions de foi montre qu’il a existé des formulations archaïques antérieures à la shahâda traditionnelle, reflet d’un monothéisme tribal très matérialiste. Quant aux demandes de pardon récurrentes dans les graffiti, elles relèveraient d’une stratégie d’écriture. Enfin, la constatation de l’absence de citation du prophète Muḥammad dans les graffiti les plus anciens montre, à elle seule, les enjeux historiques et religieux de cette recherche épigraphique.

  3. Autour d'un mystère de l'histoire du livre. Les trois versions du premier volume du Voyage pittoresque de Choiseul-Gouffier

    Directory of Open Access Journals (Sweden)

    Ioannis Koubourlis

    2009-01-01

    Full Text Available Dans cet article, il est question d'un grand mystère de l'histoire du livre, celui de l'existence de trois versions différentes du premier volume du Voyage pittoresque de la Grèce (1782 de Choiseul-Gouffier, c'est-à-dire d'un ouvrage majeur pour la floraison des idées philhellènes dans l'Europe des XVIIIe-XIXe siècles. On sait, grâce à la correspondance de l'auteur, qu'il avait pris la décision de réviser son texte dès 1783, en raison de sa candidature pour le poste d'Ambassadeur de France à Constantinople. Par contre, on n'en sait pas davantage sur le lieu et le temps exacts où il a travaillé les deux nouvelles versions, portant également la date 1782, ni d'ailleurs sur les circonstances de leur édition. Sur la base d'une étude comparative des trois versions du texte, qui met l'accent sur l'argumentation avancée chaque fois par l'auteur, nous formulons ici une série d'hypothèses pour l'interprétation de ce mystère, que nous allons examiner dans leurs détails à partir d'une étude de bibliologie qui suivra le présent article.

  4. Practical assay issues with the PERT/PBRT assay: a highly sensitive reverse transcriptase assay.

    Science.gov (United States)

    Chang, A; Dusing, S

    2006-01-01

    Product safety testing for retroviruses can be achieved by a panel of screening assays, including electron microscopy, viral gene specific PCRs, virus propagation, and detection of reverse transciptase activity. The application of PCR-based reverse transcriptase assays (PERT) that are approximately a million-fold more sensitive than conventional nucleotide incorporation assays in the testing of biologicals is described. Use of PERT assays can be applied to three areas: (i) screening for adventitious retrovirus contamination; (ii) detecting and quantifying endogenous viral particle load and (iii) monitoring levels of infectious retrovirus generation in cell lines that contain endogenous retroviruses.

  5. From Antenna to Assay

    Science.gov (United States)

    Moore, Evan G.; Samuel, Amanda P. S.; Raymond, Kenneth N.

    2009-01-01

    Conspectus Ligand-sensitized, luminescent lanthanide(III) complexes are of considerable importance because their unique photophysical properties (microsecond to millisecond lifetimes, characteristic and narrow emission bands, and large Stokes shifts) make them well suited as labels in fluorescence-based bioassays. The long-lived emission of lanthanide(III) cations can be temporally resolved from scattered light and background fluorescence to vastly enhance measurement sensitivity. One challenge in this field is the design of sensitizing ligands that provide highly emissive complexes with sufficient stability and aqueous solubility for practical applications. In this Account, we give an overview of some of the general properties of the trivalent lanthanides and follow with a summary of advances made in our laboratory in the development of highly luminescent Tb(III) and Eu(III) complexes for applications in biotechnology. A focus of our research has been the optimization of these compounds as potential commercial agents for use in Homogeneous Time-Resolved Fluorescence (HTRF) technology. Our approach involves developing high-stability octadentate Tb(III) and Eu(III) complexes that rely on all-oxygen donor atoms and using multi-chromophore chelates to increase molar absorptivity; earlier examples utilized a single pendant chromophore (that is, a single “antenna”). Ligands based on 2-hydroxyisophthalamide (IAM) provide exceptionally emissive Tb(III) complexes with quantum yield values up to ∼60% that are stable at the nanomolar concentrations required for commercial assays. Through synthetic modification of the IAM chromophore and time-dependent density functional theory (TD-DFT) calculations, we have developed a method to predict absorption and emission properties of these chromophores as a tool to guide ligand design. Additionally, we have investigated chiral IAM ligands that yield Tb(III) complexes possessing both high quantum yield values and strong

  6. Premier Hospital Quality Incentive Demonstration

    Data.gov (United States)

    U.S. Department of Health & Human Services — CMS is pursuing a vision to improve the quality of health care by expanding the information available about quality of care and through direct incentives to reward...

  7. Transporter assays and assay ontologies: useful tools for drug discovery.

    Science.gov (United States)

    Zdrazil, Barbara; Chichester, Christine; Zander Balderud, Linda; Engkvist, Ola; Gaulton, Anna; Overington, John P

    2014-06-01

    Transport proteins represent an eminent class of drug targets and ADMET (absorption, distribution, metabolism, excretion, toxicity) associated genes. There exists a large number of distinct activity assays for transport proteins, depending on not only the measurement needed (e.g. transport activity, strength of ligand–protein interaction), but also due to heterogeneous assay setups used by different research groups. Efforts to systematically organize this (divergent) bioassay data have large potential impact in Public-Private partnership and conventional commercial drug discovery. In this short review, we highlight some of the frequently used high-throughput assays for transport proteins, and we discuss emerging assay ontologies and their application to this field. Focusing on human P-glycoprotein (Multidrug resistance protein 1; gene name: ABCB1, MDR1), we exemplify how annotation of bioassay data per target class could improve and add to existing ontologies, and we propose to include an additional layer of metadata supporting data fusion across different bioassays.

  8. A novel enzyme-linked immunosorbent assay for detection of Escherichia coli O157:H7 using immunomagnetic and beacon gold nanoparticles.

    Science.gov (United States)

    Shen, Zhiqiang; Hou, Nannan; Jin, Min; Qiu, Zhigang; Wang, Jingfeng; Zhang, Bin; Wang, Xinwei; Wang, Jie; Zhou, Dongsheng; Li, Junwen

    2014-01-01

    This paper presents a functional nanoparticle-enhanced enzyme-linked immunosorbent assay (FNP-ELISA) for detection of enterohemorrhagic Escherichia coli (EHEC) O157:H7. Immunomagnetic nanoparticles (IMMPs) conjugated with monoclonal anti-O157:H7 antibody were used to capture E. coli O157:H7. Beacon gold nanoparticles (B-GNPs) coated with polyclonal anti-O157:H7 and biotin single-stranded DNA (B-DNA) were then subjective to immunoreaction with E. coli O157:H7, which was followed by streptavidin-horseradish peroxidase (Strep-HRP) conjugated with B-GNPs based on a biotin-avidin system. The solutions containing E. coli O157:H7, IMMPs, B-GNPs, and Strep-HRP were collected for detecting color change. The signal was significantly amplified with detection limits of 68 CFU mL(-1) in PBS and 6.8 × 10(2) to 6.8 × 10(3) CFU mL(-1) in the food samples. The FNP-ELISA method developed in this study was two orders of magnitude more sensitive than immunomagnetic separation ELISA (IMS-ELISA) and four orders of magnitude more sensitive than C-ELISA. The entire detection process of E. coli O157:H7 lasted only 3 h, and thus FNP-ELISA is considered as a time-saving method.

  9. MICROBIOLOGICAL ASSAY FOR VITAMIN B

    OpenAIRE

    Bishnoi Kapil*, , ,; Kataria Mahesh; Singhal Vipin; Gupta Deepika

    2012-01-01

    Micronutrients added to foods are analyzed using various procedures depending on their nature and properties. The microbiological assays are better than chemical method because any suitable change in vitamin molecule which may not be detected by chemical method will be revealed by change in microbial activity. The microbiological assay of vitamins is based upon the comparison of the stimulation of growth of bacteria by measured concentration of vitamin with that produced by known concentratio...

  10. Archéologie des monastères du premier millénaire dans le Centre-Est de la France. Conditions d’implantation et de diffusion, topographie historique et organisation

    Directory of Open Access Journals (Sweden)

    Sébastien Bully

    2009-12-01

    Full Text Available Lorsqu’en 1997 le Conseil national de la recherche archéologique publiait le bilan 1990-1994 de la recherche en France et sa nouvelle programmation, figurait parmi les thèmes prioritaires : « Les origines et les fonctions des premiers bâtiments monastiques : églises et organisation des bâtiments communautaires, relations entre architecture et règle monastique » . Il était alors rappelé que l’on ne savait rien des fondations colombaniennes et très peu des abbayes carolingiennes. Depuis lors, d...

  11. Correlation between the genotoxicity endpoints measured by two different genotoxicity assays: comet assay and CBMN assay

    Directory of Open Access Journals (Sweden)

    Carina Ladeira

    2015-06-01

    The results concerning of positive findings by micronuclei and non significant ones by comet assay, are corroborated by Deng et al. (2005 study performed in workers occupationally exposed to methotrexate, also a cytostatic drug. According to Cavallo et al. (2009, the comet assay seems to be more suitable for the prompt evaluation of the genotoxic effects, for instance, of polycyclic aromatic hydrocarbons mixtures containing volatile substances, whereas the micronucleus test seems more appropriate to evaluate the effects of exposure to antineoplastic agents. However, there are studies that observed an increase in both the comet assay and the micronucleus test in nurses handling antineoplastic drugs, although statistical significance was only seen in the comet assay, quite the opposite of our results (Maluf & Erdtmann, 2000; Laffon et al. 2005.

  12. Évolution nycthémérale des composantes biochimiques du phytoplancton de la retenue du barrage Idriss premier (Fès, Maroc)

    Science.gov (United States)

    Bahhou, J.; Alaoui Mhamdi, M.

    1999-03-01

    The diel changes of the biochemical composition of the phytoplankton were studied in the Idriss first reservoir (located on the Inaouen river at thirty Km from the city of Fes, Morrocco) during September 1994. Several biomass and metabolic indicators (proteins, carbohydrates, lipids, chlorophyll a and primary production) were assessed every fourth hour over tow days (7, 8 and 9 September). Since the Protein/Carbohydrates ratio (P/C) is largely recognised as a good integrator of the metabolic functions of the cells, we examined its distribution pattern concomitantly with aforementioned parameters. The results demonstrated enhanced P/C ratios clearly indicating that nutrients were sufficiently available for growth. In addition, this index showed a diel significant variation with levels higher in the night than in the day. Moreover, these results suggest that phytoplankton species during the night used the day-synthesised carbohydrates to insure the cell metabolic functioning. The P/C presents relatively high values in proposition to the ones that have been recorded in temperate regions, and seems to be related to azotic inputs of the Inaouen river. Dans le but de parfaire nos connaissances sur le fonctionnement du sous écosystème phytoplancton, nous nous sommes intéressés à étudier son cycle nycthéméral et son métabolisme cellulaire dans la retenue du barrage Idriss premier. Cette dernière, construite sur l'Oued Inaouène, est située à une trentaine de Km de la ville de Fès. Au cours de ce cycle, les prélèvements ont été effectués selon une séquence temporelle de 4 heures pendant 48 heures, les 7, 8 et 9 septembre 1994. L'étude de l'évolution des composantes biochimiques des cellules phytoplanctoniques à savoir les protéines, les glucides et les lipides a permis de mettre en évidence des variations nycthémérales importantes. Ces variations sont d'autant plus importantes que les variations spatiales observées entre les profondeurs. De plus

  13. Longwood Gardens——A World Premiere Horticultural Display Garden%长木公园——世界杰出的园艺展示园

    Institute of Scientific and Technical Information of China (English)

    (美)詹姆斯·哈比奇; 马琳; 常雷刚; 李星

    2011-01-01

    Longwood Gardens is recognized as one of the world premiere horticultural display gardens, inspiring people through excellence in garden design, horticulture, education and the arts. Longwood displays integrate permanent plantings and seasonal floral crops with interpretive exhibits and performing arts events, coordinated into a combined guest experience. Longwood Gardens' Horticulture Display team combines strong design principles with the unique plant pallet of over 9 500 taxa to maximize the visual impact of displays. This team often collaborates with world class designers to develop major new display themes and spaces. The intense seasonal display program is possible because of the, mostly, on-site production of over 120 000 containers of floral crops on 4 acres of dedicated production space. Longwood's team of craftsmen also support display efforts by fabricating the structural and mechanical elements often required for the elaborate design compositions. The culmination of planning, design, production, and execution come together to produce amazing horticultural displays, that are original and unique in the world, and provide wonderful memories of beauty, excitement and solitude to visitors from all over.%长木公园是世界杰出的园艺展示园之一,其精湛的花园设计、园艺、教育以及艺术品鼓舞人心.长木公园的展览将永久性栽植和时令花卉种植结合在一起,加入诠释性展览和表演艺术活动,整体协调成为一种综合性的游览体验.长木公园的园艺展示团队将强大的设计原则与9500多个独特的植物类群相结合,达到展示视觉效果的最大化.团队经常与世界一流的设计师合作来开发新的展览主题和空间.密集的季节性展览,很大程度上得益于公园的1.6万m2专用生产空间,可现场生产12万盆花卉.长木的技师团队也通过制造结构和机械元件来支持展览.顶尖的策划、设计、生产和执行结合在一起,创造出

  14. Barcoded microchips for biomolecular assays.

    Science.gov (United States)

    Zhang, Yi; Sun, Jiashu; Zou, Yu; Chen, Wenwen; Zhang, Wei; Xi, Jianzhong Jeff; Jiang, Xingyu

    2015-01-20

    Multiplexed assay of analytes is of great importance for clinical diagnostics and other analytical applications. Barcode-based bioassays with the ability to encode and decode may realize this goal in a straightforward and consistent manner. We present here a microfluidic barcoded chip containing several sets of microchannels with different widths, imitating the commonly used barcode. A single barcoded microchip can carry out tens of individual protein/nucleic acid assays (encode) and immediately yield all assay results by a portable barcode reader or a smartphone (decode). The applicability of a barcoded microchip is demonstrated by human immunodeficiency virus (HIV) immunoassays for simultaneous detection of three targets (anti-gp41 antibody, anti-gp120 antibody, and anti-gp36 antibody) from six human serum samples. We can also determine seven pathogen-specific oligonucleotides by a single chip containing both positive and negative controls.

  15. Radioreceptor assay method for insulin

    Energy Technology Data Exchange (ETDEWEB)

    Mori, K.F.; Wood, R.J. (Bureau of Drug Research, Health and Welfare Canada, Ottawa, Ontario. Health Protection Branch)

    1984-01-01

    A sensitive practical radioreceptor assay method for pharmaceutical insulin products has been developed with partially purified rat liver plasma membranes and the optimal conditions under which the best overall assay performance is obtainable have been defined. Intra- and inter-assay variations of the method averaged 7.3 and 12.2% of the man, respectively, when expressed as the coefficient of variation. Potency estimates of an insulin product obtained with the proposed method correlated well with those determined by the mouse convulsion bioassay method. Liver membranes prepared according to the method could be stored for up to ten weeks at 4/sup 0/C and for 6 months or more at -18/sup 0/C without losing insulin-binding ability.

  16. Conception et réalisation de l'unité de décision du système de déclenchement de premier niveau du détecteur LHCb au LHC

    CERN Document Server

    Laubser, Julien

    2007-01-01

    Le detecteur LHCb est l'une des quatre experiences de physique des particules installees sur la nouvelle chaine d'acceleration LHC (Large Hadron Collider) du CERN a Geneve. Afin de reduire la quantite de donnees destinees au stockage pour les analyses hors ligne, un dispositif de selection en ligne des collisions interessantes selon la physique a etudier est mis en place en parallele de la chaine d'acquisition des donnees. Ce dispositif est compose d'un premier niveau(niveau 0) realise par un systeme electronique complexe et d'un second niveau de selection realise par informatique HLT (High Level Trigger). L'unite de decision de niveau 0 (L0DU) est le systeme central du niveau 0 de declenchement. L0DU prend la decision d'accepter ou de rejeter la collision pour ce premier niveau a partir d'une fraction d'informations issues des sous-detecteurs les plus rapides (432 bits a 80 MHz). L'unite de decision est un circuit imprime 16 couches integrant des composants de haute technologie de type FPGA (Field Programmab...

  17. Study ing the influence and d imensions of Competitive Intelligence of Coaches on performance of current Soccer teams of The Iranian Premier League According to Component Balanced Scorecard (BSC

    Directory of Open Access Journals (Sweden)

    Amir Abbas BABAKIANPOUR

    2014-06-01

    Full Text Available T he aim of this study was to investigate the impact of coaches competitive intelligence on the football teams performance in the Iranian Premier League based on the Balanced Scorecard component (BSC.This prac tical research is a descriptive - correlational study in which population consisted of all head coaches and coaches from 16 Iranian Premier F ootball teams be tween 1392 - 1393. Two individual from each team were selected as coach and head coach and t he sample of 32 students were selected by census method. Questionnaire, 12 balls Fehy (2007 was used to measure competitive intelligence Coaches variables and ¬ 20 items of the questionnaire Neon (2003 was used to measure team performance. The results showed that the competitive intelligence team football coache s could affect performance and internal team performance will significantly increase. Also the team awareness c o ndition, technical knowledge, Opponents situation awareness, and awareness strategy of the coac hes had the greatest impact on team performance. In fact, coach es who having high level of competitive intellige nce will use the environmental threats as opportunities for improvement and uses it to enhance team performance.

  18. Chromosome aberration assays in Allium

    Energy Technology Data Exchange (ETDEWEB)

    Grant, W.F.

    1982-01-01

    The common onion (Allium cepa) is an excellent plant for the assay of chromosome aberrations after chemical treatment. Other species of Allium (A. cepa var. proliferum, A. carinatum, A. fistulosum and A. sativum) have also been used but to a much lesser extent. Protocols have been given for using root tips from either bulbs or seeds of Allium cepa to study the cytological end-points, such as chromosome breaks and exchanges, which follow the testing of chemicals in somatic cells. It is considered that both mitotic and meiotic end-points should be used to a greater extent in assaying the cytogenetic effects of a chemical. From a literature survey, 148 chemicals are tabulated that have been assayed in 164 Allium tests for their clastogenic effect. Of the 164 assays which have been carried out, 75 are reported as giving a positive reaction, 49 positive and with a dose response, 1 positive and temperature-related, 9 borderline positive, and 30 negative; 76% of the chemicals gave a definite positive response. It is proposed that the Allium test be included among those tests routinely used for assessing chromosomal damage induced by chemicals.

  19. 21 CFR 225.158 - Laboratory assays.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 4 2010-04-01 2010-04-01 false Laboratory assays. 225.158 Section 225.158 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) DRUGS... Laboratory assays. Where the results of laboratory assays of drug components, including assays by State...

  20. L’espace domestique au Bronze final et au premier âge du Fer dans le sud de la Corse

    Directory of Open Access Journals (Sweden)

    Peche-Quilichini, Kewin

    2015-12-01

    organización general como a nivel del espacio interno y de la arquitectura de las viviendas. [fr] L’objectif de cet article est de fournir une approche synthétique sur les formes de l’habitat en Corse au Bronze final (BF et au premier âge du Fer (F1, à l’échelle de la microrégion montagneuse de l’Alta Rocca, située dans le sud de la Corse, au coeur du bassin occidental de la Méditerranée. La problématique d’étude des sites protohistoriques non fortifiés est jeune sur l’île, mais les travaux se sont récemment multipliés et rendent compte de la complexité structurelle et évolutive des espaces habités, permettant ainsi une premiére analyse comparative. Le raisonnement s’appuie essentiellement sur l’apport des fouilles du grand habitat de Cuciurpula, initiées en 2008 et toujours en cours, ainsi que sur l’exploitation des villages de Puzzonu et de Nuciaresa, sondés en 2012. La chronologie des secteurs étudiés permet d’embrasser un arc chronologique complet entre le XIIe et le VIe siécle av. J.-C., et donc d’appréhender les phénoménes évolutifs, tant en termes d’organisation générale qu’au niveau de l’espace interne et de l’architecture des habitations.

  1. Comet Assay in Cancer Chemoprevention.

    Science.gov (United States)

    Santoro, Raffaela; Ferraiuolo, Maria; Morgano, Gian Paolo; Muti, Paola; Strano, Sabrina

    2016-01-01

    The comet assay can be useful in monitoring DNA damage in single cells caused by exposure to genotoxic agents, such as those causing air, water, and soil pollution (e.g., pesticides, dioxins, electromagnetic fields) and chemo- and radiotherapy in cancer patients, or in the assessment of genoprotective effects of chemopreventive molecules. Therefore, it has particular importance in the fields of pharmacology and toxicology, and in both environmental and human biomonitoring. It allows the detection of single strand breaks as well as double-strand breaks and can be used in both normal and cancer cells. Here we describe the alkali method for comet assay, which allows to detect both single- and double-strand DNA breaks.

  2. The skin-blanching assay.

    Science.gov (United States)

    Smit, P; Neumann, H A M; Thio, H B

    2012-10-01

    The skin-blanching assay is used for the determination and bioequivalence of dermatologic glucocorticoids (GCs). The exact mechanism of the production of blanching is not fully understood, but it is considered that local vasoconstriction of the skin microvasculature and the consequent blood-flow reduction cause this phenomenon. Several factors influence skin blanching, including drug concentration, duration of application, nature of vehicle, occlusion, posture and location. The intensity of vasoconstriction can be measured in several ways: visual or quantitative methods, such as reflectance spectroscopy, thermography, laser Doppler velocimetry and chromametry. In literature, contradicting results in the correlation of the skin-blanching assay with different tests to determine GC sensitivity have been reported, limiting its clinical usefulness.

  3. Bioluminescence assay for cell viability.

    Science.gov (United States)

    Lomakina, G Yu; Modestova, Yu A; Ugarova, N N

    2015-06-01

    Theoretical aspects of the adenosine triphosphate bioluminescence assay based on the use of the firefly luciferin-luciferase system are considered, as well as its application for assessing cell viability in microbiology, sanitation, medicine, and ecology. Various approaches for the analysis of individual or mixed cultures of microorganisms are presented, and capabilities of the method for investigation of biological processes in live cells including necrosis, apoptosis, as well as for investigation of the dynamics of metabolism are described.

  4. Protein binding assay for hyaluronate

    Energy Technology Data Exchange (ETDEWEB)

    Lacy, B.E.; Underhill, C.B.

    1986-11-01

    A relatively quick and simple assay for hyaluronate was developed using the specific binding protein, hyaluronectin. The hyaluronectin was obtained by homogenizing the brains of Sprague-Dawley rats, and then centrifuging the homogenate. The resulting supernatant was used as a source of crude hyaluronectin. In the binding assay, the hyaluronectin was mixed with (/sup 3/H)hyaluronate, followed by an equal volume of saturated (NH/sub 4/)/sub 2/SO/sub 4/, which precipitated the hyaluronectin and any (/sup 3/H)hyaluronate associated with it, but left free (/sup 3/H)hyaluronate in solution. The mixture was then centrifuged, and the amount of bound (/sup 3/H)hyaluronate in the precipitate was determined. Using this assay, the authors found that hyaluronectin specifically bound hyaluronate, since other glycosaminoglycans failed to compete for the binding protein. In addition, the interaction between hyaluronectin and hyaluronate was of relatively high affinity, and the size of the hyaluronate did not appear to substantially alter the amount of binding. To determine the amount of hyaluronate in an unknown sample, they used a competition assay in which the binding of a set amount of (/sup 3/H)hyaluronate was blocked by the addition of unlabeled hyaluronate. By comparing the degree of competition of the unknown samples with that of known amounts of hyaluronate, it was possible to determine the amount of hyaluronate in the unknowns. They have found that this method is sensitive to 1 ..mu..g or less of hyaluronate, and is unaffected by the presence of proteins.

  5. Correlation between the genotoxicity endpoints measured by two different genotoxicity assays: comet assay and CBMN assay

    OpenAIRE

    Carina Ladeira; Susana Viegas; Manuel C. Gomes

    2015-01-01

    The cytokinesis-block micronucleus cytome (CBMN) assay is a comprehensive system for measuring DNA damage; cytostasis and cytotoxicity-DNA damage events are scored specifically in once-divided binucleated cells. The endpoints possible to be measured are micronuclei (MN), a biomarker of chromosome breakage and/or whole chromosome loss, nucleoplasmic bridges (NPB), a biomarker of DNA misrepair and/or telomere end-fusions, and nuclear buds (NBUD), a biomarker of elimination of amplified DNA and/...

  6. Non-operative management of a complete anterior cruciate ligament injury in an English Premier League football player with return to play in less than 8 weeks: applying common sense in the absence of evidence.

    Science.gov (United States)

    Weiler, Richard; Monte-Colombo, Mathew; Mitchell, Adam; Haddad, Fares

    2015-04-26

    This case report illustrates and discusses the non-operative management of a complete anterior cruciate ligament (ACL) injury in an English Premier League football player, his return to play within 8 weeks and problem-free follow-up at 18 months post injury. When non-operative verses surgical ACL reconstruction is considered there are many fundamental gaps in our knowledge and currently, at elite level, there are no cases in cutting sports within the literature to guide these decisions. When the norm is for all professional footballers to be recommended surgery, it will be very challenging when circumstances and patient autonomy dictate a conservative approach, where prognosis, end points and risk are unclear and assumed to be high. This case challenges current dogma and provides a starting point for much needed debate about best practice, treatment options, research direction and not just at the elite level of sport.

  7. Injury risk factors, screening tests and preventative strategies: a systematic review of the evidence that underpins the perceptions and practices of 44 football (soccer) teams from various premier leagues.

    Science.gov (United States)

    McCall, Alan; Carling, Chris; Davison, Michael; Nedelec, Mathieu; Le Gall, Franck; Berthoin, Serge; Dupont, Gregory

    2015-05-01

    To systematically review the scientific level of evidence for the 'Top 3' risk factors, screening tests and preventative exercises identified by a previously published survey of 44 premier league football (soccer) teams. Also, to provide an overall scientific level of evidence and graded recommendation based on the current research literature. A systematic literature search (Pubmed [MEDLINE], SportDiscus, PEDRO and Cochrane databases). The quality of the articles was assessed and a level of evidence (1++ to 4) was assigned. Level 1++ corresponded to the highest level of evidence available and 4, the lowest. A graded recommendation (A: strong, B: moderate, C: weak, D: insufficient evidence to assign a specific recommendation) for use in the practical setting was given. Fourteen studies were analysed. The overall level of evidence for the risk factors previous injury, fatigue and muscle imbalance were 2++, 4 and 'inconclusive', respectively. The graded recommendation for functional movement screen, psychological questionnaire and isokinetic muscle testing were all 'D'. Hamstring eccentric had a weak graded 'C' recommendation, and eccentric exercise for other body parts was 'D'. Balance/proprioception exercise to reduce ankle and knee sprain injury was assigned a graded recommendation 'D'. The majority of perceptions and practices of premier league teams have a low level of evidence and low graded recommendation. This does not imply that these perceptions and practices are not important or not valid, as it may simply be that they are yet to be sufficiently validated or refuted by research. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

  8. Injury risk factors, screening tests and preventative strategies: a systematic review of the evidence that underpins the perceptions and practices of 44 football (soccer) teams from various premier leagues

    Science.gov (United States)

    McCall, Alan; Carling, Chris; Davison, Michael; Nedelec, Mathieu; Le Gall, Franck; Berthoin, Serge; Dupont, Gregory

    2015-01-01

    Purpose To systematically review the scientific level of evidence for the ‘Top 3’ risk factors, screening tests and preventative exercises identified by a previously published survey of 44 premier league football (soccer) teams. Also, to provide an overall scientific level of evidence and graded recommendation based on the current research literature. Methods A systematic literature search (Pubmed [MEDLINE], SportDiscus, PEDRO and Cochrane databases). The quality of the articles was assessed and a level of evidence (1++ to 4) was assigned. Level 1++ corresponded to the highest level of evidence available and 4, the lowest. A graded recommendation (A: strong, B: moderate, C: weak, D: insufficient evidence to assign a specific recommendation) for use in the practical setting was given. Results Fourteen studies were analysed. The overall level of evidence for the risk factors previous injury, fatigue and muscle imbalance were 2++, 4 and ‘inconclusive’, respectively. The graded recommendation for functional movement screen, psychological questionnaire and isokinetic muscle testing were all ‘D’. Hamstring eccentric had a weak graded ‘C’ recommendation, and eccentric exercise for other body parts was ‘D’. Balance/proprioception exercise to reduce ankle and knee sprain injury was assigned a graded recommendation ‘D’. Conclusions The majority of perceptions and practices of premier league teams have a low level of evidence and low graded recommendation. This does not imply that these perceptions and practices are not important or not valid, as it may simply be that they are yet to be sufficiently validated or refuted by research. PMID:25576530

  9. Monsieur Etienne Blanc Premier vice-président de la Région Auvergne-Rhône-Alpes Délégué aux finances, à l'administration générale, aux économies budgétaires et aux politiques transfrontalières

    CERN Multimedia

    Bennett, Sophia Elizabeth

    2017-01-01

    Monsieur Etienne Blanc Premier vice-président de la Région Auvergne-Rhône-Alpes Délégué aux finances, à l'administration générale, aux économies budgétaires et aux politiques transfrontalières

  10. Édition, diffusion et réception des premiers ouvrages sur le commerce et la comptabilité en Russie au XVIIIe siècle

    Directory of Open Access Journals (Sweden)

    Natalia Platonova

    2010-12-01

    Full Text Available À partir du XVIe siècle, des ouvrages destinés à aider les négociants dans leurs pratiques commerciales et en particulier à leur apprendre les techniques de la comptabilité circulent à travers l’Europe. Il faut attendre la seconde moitié du XVIIIe siècle pour qu’ils apparaissent en Russie. En prenant la mesure du rôle grandissant des marchands dans l’État et la société, la monarchie tenta d’organiser l’enseignement commercial et favorisa la diffusion de cette littérature. Ces premiers écrits, leur contenu, les conditions de leur diffusion, leur impact sur l’instruction et les pratiques des marchands de cette époque font l’objet de la présente étude. Cela donne l’occasion de souligner la parenté des publications russes avec l’ensemble des ouvrages de même nature imprimés en Europe occidentale entre les XVIe et XVIIIe siècles. En effet, leur parution ouvre la voie à la pénétration en Russie des savoirs et des pratiques de la comptabilité moderne. Toutefois, cette littérature n’a pas capté l’intérêt immédiat des milieux marchands russes. Ceux-ci se révèlent majoritairement indifférents à la tenue des livres en partie double et restent attachés aux modes traditionnels de formation professionnelle et de gestion des affaires. En cause, le caractère à la fois novateur et complexe du contenu de ces ouvrages, mais aussi les conditions générales dans lesquelles se déroulait l’exercice du commerce à cette époque et l’identité sociale des marchands. C’est pourquoi l’impact de la littérature commerciale et comptable du XVIIIe siècle sur les pratiques marchandes ne doit pas être surestimé. Cette littérature a surtout marqué la pensée comptable dont le développement s’amorce à partir de cette époque en Russie.  Since the sixteenth century, books written for to help the merchants in their practices and in particular to learn how to keep accounts spread across Europe. Such

  11. 21 CFR 864.7525 - Heparin assay.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Heparin assay. 864.7525 Section 864.7525 Food and... HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7525 Heparin assay. (a) Identification. A heparin assay is a device used to determine the level of the anticoagulant heparin in the...

  12. A colorimetric assay for cytokinin oxidase.

    Science.gov (United States)

    Libreros-Minotta, C A; Tipton, P A

    1995-11-01

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

  13. Multicentre comparison of a diagnostic assay

    DEFF Research Database (Denmark)

    Waters, Patrick; Reindl, Markus; Saiz, Albert;

    2016-01-01

    ) assays in neuromyelitis optica spectrum disorders (NMOSD). METHODS: Coded samples from patients with neuromyelitis optica (NMO) or NMOSD (101) and controls (92) were tested at 15 European diagnostic centres using 21 assays including live (n=3) or fixed cell-based assays (n=10), flow cytometry (n=4...

  14. 21 CFR 866.3210 - Endotoxin assay.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Endotoxin assay. 866.3210 Section 866.3210 Food... DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3210 Endotoxin assay. (a) Identification. An endotoxin assay is a device that uses serological techniques in whole blood. The device...

  15. Capillary Electrophoresis Analysis of Conventional Splicing Assays

    DEFF Research Database (Denmark)

    de Garibay, Gorka Ruiz; Acedo, Alberto; García-Casado, Zaida;

    2014-01-01

    of these assays is often challenging. Here, we explore this issue by conducting splicing assays in 31 BRCA2 genetic variants. All variants were assessed by RT-PCR followed by capillary electrophoresis and direct sequencing. If assays did not produce clear-cut outputs (Class-2 or Class-5 according to analytical...

  16. Steroid assays in paediatric endocrinology.

    Science.gov (United States)

    Honour, John W

    2010-01-01

    Most steroid disorders of the adrenal cortex come to clinical attention in childhood and in order to investigate these problems, there are many challenges to the laboratory which need to be appreciated to a certain extent by clinicians. The analysis of sex steroids in biological fluids from neonates, over adrenarche and puberty present challenges of specificities and concentrations often in small sample sizes. Different reference ranges are also needed for interpretations. For around 40 years, quantitative assays for the steroids and their regulatory peptide hormones have been possible using immunoassay techniques. Problems are recognised and this review aims to summarise the benefits and failings of immunoassays and introduce where tandem mass spectrometry is anticipated to meet the clinical needs for steroid analysis in paediatric endocrine investigations. It is important to keep a dialogue between clinicians and the laboratory, especially when any laboratory result does not make sense in the clinical investigation.

  17. Predictive Assay For Cancer Targets

    Energy Technology Data Exchange (ETDEWEB)

    Suess, A; Nguyen, C; Sorensen, K; Montgomery, J; Souza, B; Kulp, K; Dugan, L; Christian, A

    2005-09-19

    Early detection of cancer is a key element in successful treatment of the disease. Understanding the particular type of cancer involved, its origins and probable course, is also important. PhIP (2-amino-1-methyl-6 phenylimidazo [4,5-b]pyridine), a heterocyclic amine produced during the cooking of meat at elevated temperatures, has been shown to induce mammary cancer in female, Sprague-Dawley rats. Tumors induced by PhIP have been shown to contain discreet cytogenetic signature patterns of gains and losses using comparative genomic hybridization (CGH). To determine if a protein signature exists for these tumors, we are analyzing expression levels of the protein products of the above-mentioned tumors in combination with a new bulk protein subtractive assay. This assay produces a panel of antibodies against proteins that are either on or off in the tumor. Hybridization of the antibody panel onto a 2-D gel of tumor or control protein will allow for identification of a distinct protein signature in the tumor. Analysis of several gene databases has identified a number of rat homologs of human cancer genes located in these regions of gain and loss. These genes include the oncogenes c-MYK, ERBB2/NEU, THRA and tumor suppressor genes EGR1 and HDAC3. The listed genes have been shown to be estrogen-responsive, suggesting a possible link between delivery of bio-activated PhIP to the cell nucleus via estrogen receptors and gene-specific PhIP-induced DNA damage, leading to cell transformation. All three tumors showed similar silver staining patterns compared to each other, while they all were different than the control tissue. Subsequent screening of these genes against those from tumors know to be caused by other agents may produce a protein signature unique to PhIP, which can be used as a diagnostic to augment optical and radiation-based detection schemes.

  18. Activity assay of membrane transport proteins

    Institute of Scientific and Technical Information of China (English)

    Hao Xie

    2008-01-01

    Membrane transport proteins are integral membrane proteins and considered as potential drug targets. Activity assay of transport proteins is essential for developing drugs to target these proteins. Major issues related to activity assessment of transport proteins include availability of transporters,transport activity of transporters, and interactions between ligands and transporters. Researchers need to consider the physiological status of proteins (bound in lipid membranes or purified), availability and specificity of substrates, and the purpose of the activity assay (screening, identifying, or comparing substrates and inhibitors) before choosing appropriate assay strategies and techniques. Transport proteins bound in vesicular membranes can be assayed for transporting substrate across membranes by means of uptake assay or entrance counterflow assay. Alternatively, transport proteins can be assayed for interactions with ligands by using techniques such as isothermal titration calorimetry, nuclear magnetic resonance spectroscopy, or surface plasmon resonance. Other methods and techniques such as fluorometry, scintillation proximity assay, electrophysiological assay, or stopped-flow assay could also be used for activity assay of transport proteins. In this paper the major strategies and techniques for activity assessment of membrane transport proteins are reviewed.

  19. Mise en œuvre et premiers effets d’un projet d’agrégation céréalier dans la région de Bni Saden (province de Séfrou

    Directory of Open Access Journals (Sweden)

    Khawla Hdidi

    2015-10-01

    Full Text Available Les premiers projets de développement élaborés dans le cadre du Plan Maroc Vert ont été initiés en 2010. L’étude présente une analyse de la mise en oeuvre et des premiers effets d’un de ces projets. Il s’agit d’un projet d’agrégation, relevant du Pilier I, dans la filière céréalière de la province de Séfrou. Des entretiens ont été effectués auprès de 25 agriculteurs du projet et 25 agriculteurs de zones témoins situées aux alentours de la zone du projet. L’étude des effets a été faite en qualifiant comment les agriculteurs agrégés auraient évolué sans projet. Ceci a été effectué par la méthode d’appariement (matching, en associant à chaque agriculteur du projet les agriculteurs hors projet qui avaient, avant projet, certaines caractéristiques identiques à cet agriculteur. Le projet a consisté en pratique en des formations, une mise à disposition d’intrants et un taux amélioré de subvention pour l’acquisition de matériel agricole. Ces activités ont conjointement permis d’accélérer l’amélioration et l’intensification des pratiques de production des céréales et une amélioration des rendements. Les agriculteurs du projet communiquent plus entre eux sur leurs pratiques agricoles. Cependant, l’association qui regroupe les agriculteurs du projet ne joue encore qu’un rôle limité de coordination et de négociation avec les acteurs extérieurs, et en particulier avec l’agrégateur.

  20. Les nécropoles à incinérations gallo-romaines du grand-duché de Luxembourg- Premiers résultats d'une recherche en cours

    Directory of Open Access Journals (Sweden)

    Michel Polfer

    1997-09-01

    Full Text Available La contribution vise à présenter les premiers résultats d'un projet de recherche dont les buts sont situés sur deux plans: ◦l'élaboration d'un corpus aussi complet que possible de toutes les traces archéologiques de tombes individuelles et de nécropoles gallo-romaines, y compris les monuments et les inscriptions funéraires ◦l'analyse archéologique proprement dite des structures et des objets (typologie, chronologie etc. ainsi que l'étude des phénomènes religieux, culturels et sociaux qui transparaissent à travers ceux-ci. Le cadre géographique choisi est celui du Grand-Duché de Luxembourg actuel, qui représente une partie importante de l'ancienne civitas treverorum. D'un point de vue chronologique, toutes les découvertes archéologiques datables entre la deuxième moitié du 1er siècle ap. J.-C et le 5e siècle ap.- J.-C. sont pris en compte. Après un aperçu sur l'histoire de la recherche archéologique funéraire gallo-romaine au Luxembourg, l'état actuel du corpus contenant un total de 300 sites différents est présenté à l'aide de cartes et d'une banque de données. Sont ensuite présentés les premiers résultats de l'enquête relatifs à ◦la répartition géographique des nécropoles ◦les relations entre l'habitat rural, les nécropoles et les monuments funéraires ◦l' organisation spatio-temporelle des nécropoles rurales ◦les pratiques dépositionnelles et la hiérarchisation sociale au sein des nécropoles ◦les différents modes cinéraires et les rapports entre incinération et inhumation

  1. Les nécropoles à incinérations gallo-romaines du grand-duché de Luxembourg- Premiers résultats d'une recherche en cours

    Directory of Open Access Journals (Sweden)

    Michel Polfer

    1997-12-01

    Full Text Available La contribution vise à présenter les premiers résultats d'un projet de recherche dont les buts sont situés sur deux plans: l'élaboration d'un corpus aussi complet que possible de toutes les traces archéologiques de tombes individuelles et de nécropoles gallo-romaines, y compris les monuments et les inscriptions funéraires l'analyse archéologique proprement dite des structures et des objets (typologie, chronologie etc. ainsi que l'étude des phénomènes religieux, culturels et sociaux qui transparaissent à travers ceux-ci. Le cadre géographique choisi est celui du Grand-Duché de Luxembourg actuel, qui représente une partie importante de l'ancienne civitas treverorum. D'un point de vue chronologique, toutes les découvertes archéologiques datables entre la deuxième moitié du 1er siècle ap. J.-C et le 5e siècle ap.- J.-C. sont pris en compte. Après un aperçu sur l'histoire de la recherche archéologique funéraire gallo-romaine au Luxembourg, l'état actuel du corpus contenant un total de 300 sites différents est présenté à l'aide de cartes et d'une banque de données. Sont ensuite présentés les premiers résultats de l'enquête relatifs à -la répartition géographique des nécropoles -les relations entre l'habitat rural, les nécropoles et les monuments funéraires -l' organisation spatio-temporelle des nécropoles rurales -les pratiques dépositionnelles et la hiérarchisation sociale au sein des nécropoles -les différents modes cinéraires et les rapports entre incinération et inhumation

  2. Serum tau protein as a marker of disease activity in enterohemorrhagic Escherichia coli O111-induced hemolytic uremic syndrome.

    Science.gov (United States)

    Kuroda, Mondo; Shimizu, Masaki; Inoue, Natsumi; Ikeno, Iku; Nakagawa, Hiroyasu; Yokoi, Ayano; Niida, Yo; Konishi, Michio; Kaneda, Hisashi; Igarashi, Noboru; Yamahana, Junya; Taneichi, Hiromichi; Kanegane, Hirokazu; Ito, Mika; Saito, Shigeru; Furuichi, Kengo; Wada, Takashi; Nakagawa, Masaru; Yokoyama, Hitoshi; Yachie, Akihiro

    2015-01-01

    Tau protein levels in cerebrospinal fluid (CSF) and serum are elevated in patients with various central nervous system diseases. We investigated whether serum tau protein levels are useful for predicting and assessing disease activity of acute encephalopathy (AE) in enterohemorrhagic Escherichia coli (EHEC) O111-induced hemolytic uremic syndrome (HUS; EHEC encephalopathy). Serum samples were obtained from 14 patients with EHEC O111/HUS, 20 patients with non-EHEC-related AE, and 20 age- and sex-matched healthy controls. CSF samples were obtained from 2 patients with EHEC encephalopathy and 20 patients with non-EHEC-related AE. Tau protein levels and levels of several proinflammatory cytokines were quantified by enzyme-linked immunosorbent assays. Results were compared with the clinical features of EHEC encephalopathy, including magnetic resonance image (MRI) findings. Serum tau levels in patients with EHEC encephalopathy were significantly elevated compared with those in patients with EHEC O111/HUS without encephalopathy, patients with non-EHEC-related AE, and healthy controls. The ratio of CSF tau levels to serum tau levels was >1.0 in all patients with non-EHEC-related AE but encephalopathy. Serum tau protein levels increased rapidly and markedly in patients with severe EHEC 0111/HUS and encephalopathy when HUS occurred, but were not elevated in mild patients, even in the HUS phase. Furthermore, changes in serum tau protein levels in patients with EHEC encephalopathy were consistent with abnormalities on brain MRI and were positively correlated with proinflammatory cytokine levels. Our results indicate that serum tau protein might be useful to predict and assess disease activity of EHEC encephalopathy. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Development and application of assays for serotonin

    Energy Technology Data Exchange (ETDEWEB)

    Gow, I.F.

    1987-01-01

    In this thesis, two assays for serotonin were developed, validated, and used to investigate the relationship between platelet aggregation, serotonin levels and sodium status and serotonin levels and platelet function in patients with cardiovascular disease. A radioimmunoassay (RIA) using an (/sup 125/I)-labelled tracer was developed and validated for the measurement of serotonin in human platelet-rich plasma (PRP) and rat serum. Antisera were raised against N-succinamylserotonin conjugated to bovine albumin and, to improve assay sensitivity, the analyte was made chemically similar to the immunogen by conversion to N-acetylserotonin prior to assay, using the specific amino reagent N-acetoxysuccinimide. An assay for serotonin using high-pressure liquid chromatography with electrochemical detection (HPLC-ECD) was developed, and used to validate the RIA. The RIA can be used to assay up to 100 samples/day compared with 10-20/day by the HPLC-ECD assay.

  4. Direct Spectrophotometric Assay for Benzaldehyde Lyase Activity

    Directory of Open Access Journals (Sweden)

    Dessy Natalia

    2011-01-01

    Full Text Available Benzaldehyde lyase from Pseudomonas fluorescens Biovar I. (BAL, EC 4.1.2.38 is a versatile catalyst for the organic synthesis of chiral α-hydroxy ketones. To allow fast assessment of enzyme activity, a direct spectrophotometric assay is desirable. Here, a new robust and easy-to-handle assay based on UV absorption is presented. The assay developed is based on the ligation of the α-hydroxy ketone (R-2,2′-furoin from 2-furaldehyde. A robust assay with direct monitoring of the product is facilitated with a convenient concentration working range minimising experimental associated with low concentrations.

  5. Approche dynamique du premier harmonique pour la modélisation de convertisseurs AC-AC à étage intermédiaire continu. Application aux générateurs à induction

    Science.gov (United States)

    Bacha, S.; Rognon, J. P.; Ferrieux, J. P.; Bendaas, M. L.

    1995-02-01

    In this paper, we present a modelling technique for power electronics converters with both DC and AC stages. This technique is based on a dynamical approach of the first harmonic method. The approach is first applied to an idealized converter and second is extended to a framework working under discontinuous conduction mode. At the end, comparative simulations are done to validate the continuous built model. Dans cet article, il est présenté une technique de modélisation de convertisseurs présentant à la fois des étages continus et alternatifs. Cette technique est basée sur une approche dynamique de la méthode du premier harmonique. La technique est tout d'abord appliquée à un convertisseur idéalisé pour être ensuite étendue à une structure travaillant en conclusion discontinue. En dernier lieu, des simulations viennent valider le modèle continu mis au point.

  6. The Barclays Premier League game field emergency predicting and early warning mechanism and its inspiration to China%英超赛场突发事件预测预警机制及对我国的启示

    Institute of Scientific and Technical Information of China (English)

    刘亚云; 罗恒; 钟丽萍

    2014-01-01

    The authors gave a detailed introduction to the risk assessing, monitoring and early warning mechanism in the Barclays Premier League game field emergency predicting and early warning mechanism as well as gold, sil-ver and bronze levels of emergency handling. The inspiration gotten therefrom is that setting up a dedicated risk management organization, cultivating risk management professionals, strengthening the construction of the predict-ing and early warning system, and establishing a sports game emergency response command mechanism, are effec-tive ways to prevent sports game field emergences and reduce losses caused by such emergencies.%通过对英超赛场突发事件预测预警机制的风险评估、监测预警机制以及“金、银、铜”3级应急处置等进行了详细介绍。从中得到的启示是设置专门的风险管理机构,培养专业风险管理人才;加强预测预警系统的建设;建立体育赛事突发事件应急指挥机制,是预防体育赛场突发事件以及降低突发事件造成损失的有效途径。

  7. Assay-dependent variability of serum insulin concentrations: a comparison of eight assays.

    Science.gov (United States)

    Tohidi, Maryam; Arbab, Parvaneh; Ghasemi, Asghar

    2017-04-01

    Although insulin measurement is essential for both clinical and research purposes, there is currently no reference method for insulin assays. The aim of this study was to compare results of serum insulin determined by a number of commercially available assays. We compared eight insulin assays by analyzing 165 serum samples. Assays included two chemiluminescence (Roche and DiaSorin), four ELISA (Tosoh, Mercodia, Monobind, and Diametra), and two IRMA (Izotop and BioSource) methods. Each assay was compared with the mean of all assay methods and Bland-Altman difference plots were used to measure agreement between each assay and overall mean. Least squared perpendicular distance regression analysis (Deming's method) was used to calculate slope and intercept for bias and also for each assay vs. mean of eight assays. Findings showed that the lowest and highest median insulin concentrations varied by a factor of 1.8. Maximum and minimum correlations with mean of assays were observed for Roche (0.992) and BioSource (0.844), respectively. Significant bias was observed in six assays. In pairwise comparisons of different assays, the highest and least mean differences were 7.78 μU/mL and -0.14 μU/mL, respectively. In conclusion, serum insulin measurement with different assays showed a maximum of 1.8-fold difference, a point that should be taken into consideration in the interpretation of circulating insulin levels in both clinical and research fields.

  8. METHODOLOGICAL ASPECTS OF QUANTITATIVE RECEPTOR ASSAYS

    NARCIS (Netherlands)

    SMISTEROVA, J; ENSING, K; DEZEEUW, RA

    1994-01-01

    Receptor assays occupy a particular position in the methods used in bioanalysis, as they do not exploit the physico-chemical properties of the analyte. These assays make use of the property of the analyte to bind to the specific binding site (receptor) and to competitively replace a labelled ligand

  9. Assessing sediment contamination using six toxicity assays

    Directory of Open Access Journals (Sweden)

    Allen G. BURTON Jr.

    2001-08-01

    Full Text Available An evaluation of sediment toxicity at Lake Orta, Italy was conducted to compare a toxicity test battery of 6 assays and to evaluate the extent of sediment contamination at various sediment depths. Lake Orta received excessive loadings of copper and ammonia during the 1900’s until a large remediation effort was conducted in 1989-90 using lime addition. Since that time, the lake has shown signs of a steady recovery of biological communities. The study results showed acute toxicity still exists in sediments at a depth of 5 cm and greater. Assays that detected the highest levels of toxicity were two whole sediment exposures (7 d using Hyalella azteca and Ceriodaphnia dubia. The MicrotoxR assay using pore water was the third most sensitive assay. The Thamnotox, Rototox, Microtox solid phase, and Seed Germination-Root Elongation (pore and solid phase assays showed occasional to no toxicity. Based on similarity of responses and assay sensitivity, the two most useful assays were the C. dubia (or H. azteca and Microtox pore water. These assays were effective at describing sediment toxicity in a weight-of-evidence approach.

  10. Radioreceptor assay: theory and applications to pharmacology

    Energy Technology Data Exchange (ETDEWEB)

    Perret, G. (U.E.R. de Medecine, Sante et Biologie Humaine, 93 - Bobigny (France)); Simon, P. (Faculte de Medecine Pitie-Salpetriere, 75 - Paris (France))

    The aim of the first part of this work is to present the theory of the radioreceptor assay and to compare it to the other techniques of radioanalysis (radioimmunoassay, competitive protein binding assays). The technology of the radioreceptor assay is then presented and its components (preparation of the receptors, radioligand, incubation medium) are described. The analytical characteristics of the radioreceptor assay (specificity, sensitivity, reproductibility, accuracy) and the pharmacological significance of the results are discussed. The second part is devoted to the description of the radioreceptor assays of some pharmacological classes (neuroleptics, tricyclic antidepressants, benzodiazepines, ..beta..-blockers, anticholinergic drugs) and to their use in therapeutic drug monitoring. In conclusion, by their nature, radioreceptor assays are highly sensitive, reliable, precise, accurate and simple to perform. Their chief disadvantage relates to specificity, since any substance having an appreciable affinity to the receptor site will displace the specifically bound radioligand. Paradoxically in some cases, this lack of specificity may be advantageous in that it allows for the detection of not only the apparent compound but of active metabolites and endogenous receptor agonists as well and in that radioreceptors assays can be devised for a whole pharmacological class and not only for one drug as it is the case for classical physico-chemical techniques. For all these reasons future of radioreceptor assay in pharmacology appears promising.

  11. A Continuous, Fluorogenic Sirtuin 2 Deacylase Assay

    DEFF Research Database (Denmark)

    Galleano, Iacopo; Schiedel, Matthias; Jung, Manfred

    2016-01-01

    and kinetic insight regarding sirtuin inhibitors, it is important to have access to efficient assays. In this work, we report readily synthesized fluorogenic substrates enabling enzyme-economical evaluation of SIRT2 inhibitors in a continuous assay format as well as evaluation of the properties of SIRT2...

  12. Acellular comet assay: a tool for assessing variables influencing the alkaline comet assay.

    Science.gov (United States)

    Kennedy, Erin K; McNamee, James P; Prud'homme Lalonde, Louise; Jones, Trevor; Wilkinson, Diana

    2012-01-01

    In this study, an acellular modification to the alkaline comet assay to further evaluate key variables within the assay that may influence the outcome of genotoxicity studies is described. This acellular comet assay can detect differences of 0.2 Gy of (60)Co gamma-ray radiation between 0 and 1 Gy and differences of 1 Gy between 0 and 8 Gy; thus, this assay is applicable for a wide range of DNA damage levels. It is also shown that DNA damage from different radiation energies was not significantly different from (60)Co gamma-ray. This assay displayed a statistical increase in DNA damage due to uncontrolled exposure to natural light; however, the slope of the dose-response curve for light-exposed samples was similar to that for samples protected from light. A comparison of the alkaline comet assay with the acellular comet assay allowed for the intrinsic repair capacity of the alkaline comet assay to be quantified.

  13. Assays for Determination of Protein Concentration.

    Science.gov (United States)

    Olson, Bradley J S C

    2016-06-01

    Biochemical analysis of proteins relies on accurate quantification of protein concentration. Detailed in this appendix are some commonly used methods for protein analysis, e.g., Lowry, Bradford, bicinchoninic acid (BCA), UV spectroscopic, and 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) assays. The primary focus of this report is assay selection, emphasizing sample and buffer compatibility. The fundamentals of generating protein assay standard curves and of data processing are considered, as are high-throughput adaptations of the more commonly used protein assays. Also included is a rapid, inexpensive, and reliable BCA assay of total protein in SDS-PAGE sample buffer that is used for equal loading of SDS-PAGE gels. © 2016 by John Wiley & Sons, Inc.

  14. Improved benzodiazepine radioreceptor assay using the MultiScreen (R) Assay System

    NARCIS (Netherlands)

    Janssen, MJ; Ensing, K; de Zeeuw, RA

    1999-01-01

    In this article, an improved benzodiazepine radioreceptor assay is described, which allows substantial reduction in assay time, The filtration in this method was performed by using the MultiScreen(R) Assay System. The latter consists of a 96-well plate with glass fibre filters sealed at the bottom,

  15. Nano-immunosafety: issues in assay validation

    Energy Technology Data Exchange (ETDEWEB)

    Boraschi, Diana; Italiani, Paola [Institute of Biomedical Technologies, National Research Council, Via G. Moruzzi 1, 56124 Pisa (Italy); Oostingh, Gertie J; Duschl, Albert [Department of Molecular Biology, University of Salzburg, Hellbrunnerstrasse 34, 5020 Salzburg (Austria); Casals, Eudald; Puntes, Victor F [Institut Catala de Nanotecnologia, Campus de la UAB - Facultat de Ciencies, Edifici CM7, 08193 Bellaterra (Spain); Nelissen, Inge, E-mail: diana.boraschi@itb.cnr.it [VITO NV, Boeretang 200, BE-2400 Mol (Belgium)

    2011-07-06

    Assessing the safety of engineered nanomaterials for human health must include a thorough evaluation of their effects on the immune system, which is responsible for defending the integrity of our body from damage and disease. An array of robust and representative assays should be set up and validated, which could be predictive of the effects of nanomaterials on immune responses. In a trans-European collaborative work, in vitro assays have been developed to this end. In vitro tests have been preferred for their suitability to standardisation and easier applicability. Adapting classical assays to testing the immunotoxicological effects of nanoparticulate materials has raised a series of issues that needed to be appropriately addressed in order to ensure reliability of results. Besides the exquisitely immunological problem of selecting representative endpoints predictive of the risk of developing disease, assay results turned out to be significantly biased by artefactual interference of the nanomaterials or contaminating agents with the assay protocol. Having addressed such problems, a series of robust and representative assays have been developed that describe the effects of engineered nanoparticles on professional and non-professional human defence cells. Two of such assays are described here, one based on primary human monocytes and the other employing human lung epithelial cells transfected with a reporter gene.

  16. Mobile non-destructive assay system

    Energy Technology Data Exchange (ETDEWEB)

    Colarusso, A.P.; Audas, J.H.; Bieri, J.M.; Herrera, G.C.; Hastings, R.D.; Horton, W.S.; Kuckertz, T.H.; Kunz, W.E.; Medvick, P.A.; Vogel, P.A.

    1987-07-01

    A mobile system for non-destructive assay (NDA), developed at the Los Alamos National Laboratory, provides accurate and sensitive measurements for transuranic (TRU) isotopes contained in 208-iota drums of miscellaneous nuclear wastes. The NDA unit consists of four major subsystems: an assay chamber, counting and digital electronics, data acquisition, and a neutron generator. It performs both active and passive neutron waste measurements. The former determines the amount of fissile isotopes at a sensitivity level of 1 mg plutonium. The latter determines spontaneous fission and ..cap alpha..,n) isotopes at a comparable level. A complete assay consists of sequential active and passive measurements. The assay measurement and other supporting data are incorporated in a commercial spreadsheet program (Lotus 1,2,3) for further analysis, which includes various matrix corrections and a determination of whether or not the drum exceeds the 100-nCi/g threshold for TRU wastes. Field tests have been performed on three separate occasions, accomplishing more than 1800 waste drum assays. These waste drum assays are discussed, especially those comparing passive and active neutron measurements with independent segmented gamma scan assays. Results obtained with a set of 15 drums containing plutonium prepared from standards and actual hot waste matrices are also reviewed.

  17. Analytical characterization of the APTIMA HPV Assay.

    Science.gov (United States)

    Dockter, Janel; Schroder, Astrid; Eaton, Barbara; Wang, Ann; Sikhamsay, Nathan; Morales, Liezel; Giachetti, Cristina

    2009-07-01

    Human papillomavirus (HPV) testing has improved the sensitivity for the detection of cervical pre-cancer and cancer as compared to Pap testing. Several HPV tests are commercially available and most target the DNA from 13 or 14 high-risk HPV types. The APTIMA HPV Assay however, detects HPV E6/E7 mRNA from 14 high-risk types of HPV: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68. To determine the analytical performance characteristics of the APTIMA HPV Assay. Analytical sensitivity, analytical specificity, reproducibility, and the effect of potentially interfering substances was determined for the APTIMA HPV Assay on both the DTS (semi-automated) and TIGRIS DTS (fully automated) systems. The 95% detection limit for both systems was between 17 and 488 copies/reaction, depending on the HPV type. The assay did not cross-react with normal flora and opportunistic organisms that may be found in cervical samples, or low-risk HPV types. Spermicides, anti-fungal and anti-itch medications, whole blood, glacial acetic acid, and most lubricants did not interfere with assay performance. Those lubricants containing polyquaternium 15 did interfere with assay performance. Inter-instrument, inter-operator, inter-lot, and inter-run signal variability were 99% of the data. Intra-run variability was HPV Assay showed excellent performance and robustness.

  18. Polymerase chain reaction assay for avian polyomavirus.

    OpenAIRE

    Phalen, D.N.; Wilson, V G; Graham, D L

    1991-01-01

    A polymerase chain reaction assay was developed for detection of budgerigar fledgling disease virus (BFDV). The assay used a single set of primers complementary to sequences located in the putative coding region for the BFDV VP1 gene. The observed amplification product had the expected size of 550 bp and was confirmed to derive from BFDV DNA by its restriction digestion pattern. This assay was specific for BFDV and highly sensitive, being able to detect as few as 20 copies of the virus. By us...

  19. Premiers éléments de caractérisation génétique de l'alose du Rhône (Alosa fallax rhodanensis Roule, 1924

    Directory of Open Access Journals (Sweden)

    LE CORRE M.

    1998-07-01

    Full Text Available La systématique des deux espèces du genre Alosa (A. alosa Linné, 1758-grande alose- et A. fallax Lacépède, 1803 -alose feinte- colonisant l'Atlantique-est et la Méditerranée reste encore confuse en l'absence de caractérisation génétique. En particulier, six sous-espèces ont été reconnues chez l'alose feinte uniquement à partir de critères morphométriques et méristiques. Parmi ces sous-espèces, l'une d'entre elles (Alosa fallax rhodanensis Roule, 1924 -alose du Rhône- est inféodée au bassin méditerranéen français et notamment au fleuve Rhône. Les critères morphométriques et méristiques de ce taxon le rattachent sans équivoque à A. fallax. Cependant, son gabarit moyen et certaines de ses caractéristiques écologiques le rapprochent fortement d'A. alosa. Ainsi, pour connaître la véritable identité de l'alose du Rhône, une première caractérisation génétique a été entreprise à l'aide d'une analyse électrophorétique des protéines du foie (gel d'amidon et du sang (isoélectrofocalisation prélevées sur des échantillons d'adultes. Dans un premier temps, deux des loci les plus performants pour différencier les deux aloses atlantiques (MPI, Mannose Phosphate Isomérase et HBA, chaîne α de l'Hémoglobine ont été utilisés. L'observation des fréquences allèliques montre que l'alose du Rhône semble appartenir à l'espèce A. fallax. Dans un deuxième temps, un troisième locus (ADH, Alcool Déshydrogénase a été utilisé car il a été reconnu comme permettant de discriminer les populations d'alose feinte du Portugal. L'expression de sa fréquence allèlique montre que la population du Rhône resterait proche de celle des fleuves du nord du Portugal. Ces premiers résultats sont discutés en fonction des récentes études génétiques réalisées sur les aloses de l'Atlantique. Elles montrent que, compte tenu du caractère particulier de ce taxon, sa caractérisation génétique doit être compl

  20. La sodomie dans l’affaire Théophile de Viau : questions de genre et de sexualité dans la France du premier xviie siècle

    Directory of Open Access Journals (Sweden)

    Matthieu Dupas

    2010-04-01

    Full Text Available L’affaire Théophile est source de malentendus. Alors que l’historiographie du libertinage, en se concentrant sur le libertinage érudit a fini par oublier que la sodomie était un enjeu majeur du procès, les historiens de la répression homosexuelle abordent les relations sexuelles entre hommes dans une perspective essentialiste, gommant toute distinction entre sodomie au xviie siècle et homosexualité à l’époque contemporaine. Il s’agit donc la première partie de cet article de souligner que la sodomie est au cœur de l’affaire à la fois comme objet de représentation littéraire et comme pratique illicite. La seconde partie consiste dans une historicisation des discours ayant trait aux relations sexuelles entre hommes, qui montre qu’on ne saurait confondre « homosexualité masculine » et « sodomie ». Celle-ci désigne en effet des rapports anaux aussi bien entre hommes et femmes qu’entre hommes seuls ; lorsque la notion est mobilisée pour renvoyer à des relations anales entre hommes, elle ne constitue pas pour autant une orientation sexuelle ; en tant que concept issu de la théologie, elle ne relève pas de la sexualité ; aucune déviation de genre ne lui est associée. Dans une troisième partie, il s’agit de montrer que la notion de sodomie ne doit pas nous conduire à ignorer les autres discours pouvant porter sur les relations sexuelles entre hommes dans la France du premier xviie siècle et qu’à ce titre, on ne saurait se suffire de cette catégorie pour aborder la question. Bref, cet article a pour but de poser les premiers jalons d’une histoire de l’homosexualité masculine dans la France du xviie siècle d’un point de vue historiciste, sinon constructionniste.Sodomy in the Theophile de Viau affair: questions of gender and sexuality in early modern France.There is a puzzling misunderstanding about the Theophile affair. While the historiography of libertinism, focusing on erudite

  1. Determination of cell survival after irradiation via clonogenic assay versus multiple MTT Assay - A comparative study

    Directory of Open Access Journals (Sweden)

    Buch Karl

    2012-01-01

    Full Text Available Abstract For studying proliferation and determination of survival of cancer cells after irradiation, the multiple MTT assay, based on the reduction of a yellow water soluble tetrazolium salt to a purple water insoluble formazan dye by living cells was modified from a single-point towards a proliferation assay. This assay can be performed with a large number of samples in short time using multi-well-plates, assays can be performed semi-automatically with a microplate reader. Survival, the calculated parameter in this assay, is determined mathematically. Exponential growth in both control and irradiated groups was proven as the underlying basis of the applicability of the multiple MTT assay. The equivalence to a clonogenic survival assay with its disadvantages such as time consumption was proven in two setups including plating of cells before and after irradiation. Three cell lines (A 549, LN 229 and F 98 were included in the experiment to study its principal and general applicability.

  2. 使用i-gel(R)与La Premiere(R)一次性喉罩后咽喉与颈部并发症的比较:一项双盲随机对照试验

    Institute of Scientific and Technical Information of China (English)

    Christiaan Keijzer; 刘素平; Dirk R. Buitelaar; Katina M. Efthymiou; Michael (S)rámek; Julia ten Cate; May Ronday; Tino Stoppa; Johannes M. Huitink; Peter F. Schutte

    2011-01-01

    背景 很多声门上的气道装置都可用于气道管理.由于i-gel(R)不使用可充气的套囊,我们推测,相对于标准的喉罩(LM),使用i-gel(R)应该能减少手术后喉部与颈部的不适.方法 218例患者被随机分组,分别使用i-gel(R)或LaPremiere(R)喉罩进行气道管理,并于手术后1、24和48小时随访患者喉部与颈部的不适症状,随访者和患者对所用通气装置均不知情.结果 109例患者应用i-gel,103例患者使用La Premiere喉罩.与LM组比较,i-gel组在手术后1小时(6与32)、24小时(7与48)、48小时(5与25)咽喉疼痛的发生率明显减少;出现吞咽困难的情况两组患者相似;i-gel组在手术后24小时(1与27)、48小时(1与7)颈部疼痛的发生率也较少.结论 通过本随机研究,使用i-gel声门上装置导致的手术后喉部与颈部的不适小于LaPremiere喉罩.

  3. La fouille du fort Saint-Georges à Chinon (Indre-et-Loire. Premiers résultats The excavation of fort Saint-Georges at Chinon (Indre-et-Loire. First results

    Directory of Open Access Journals (Sweden)

    Bruno Dufaÿ

    2006-05-01

    Full Text Available Cette note présente les premiers résultats des fouilles menées en 2003 et 2004 sur la quasi-totalité du fort Saint-Georges à Chinon (Indre-et-Loire. Celui-ci est l’un des trois éléments de la forteresse médiévale qui domine la ville. La fouille a permis de préciser la fonction du fort, construit dans la deuxième moitié du XIIe s., à l’époque où Chinon est le centre administratif des possessions continentales des Plantagenêt, rois d’Angleterre. Du point de vue militaire, il formait une fortification avancée, protégeant le château principal, selon une structure que Richard Cœur de Lion appliquera au Château Gaillard. À l’intérieur, de vastes bâtiments constituaient des logis, conçus peut-être au départ pour héberger la chancellerie royale.This article presents the first results of the excavations undertaken in 2003 and 2004 over almost all of the Fort Saint-Georges at Chinon (Indre-et-Loire, one of three elements of the medieval fortress which dominates the town. The excavation enabled us to clarify the function of the fort, built in the 2nd half of the 12th century at a time when Chinon was the administrative centre of the continental possesions of the Plantagenet King of England. From a military point of view, it formed an advanced fortification protecting the main castle, within a structure that Richard the Lionheart would apply to the Chayeau Gaillard. Inside, some vast buildings made up the dwellings, designed perhaps initially to house the royal chanceller.

  4. Archéologie et sociabilité: la délégation du Pérou au Premier Congrès international des Américanistes, Nancy, 1875

    Directory of Open Access Journals (Sweden)

    1989-01-01

    Full Text Available L'auteur tente de montrer comment l'analyse de la composition de la délégation péruvienne au premier Congrès International des Américanistes peut fournir une vision différente du mouvement anthropologique au XIXème siècle, dans la mesure où la plupart des membres de cette délégation étaient peut-être plus préoccupés de représentation que de l'avancement des sciences humaines. D'où l'idée d'un intérêt pour l'archéologie considéré alors comme un acte de sociabilité. El autor intenta demostrar que el análisis de la delegación peruana al primer Congreso Internacional de Americanistas puede dar otra visión del movimiento antropológico en el siglo XIX, en la medida en que la mayoría de los miembros de esta delegación estaban tal vez más preocupados por su imagen pública que por el progreso de las ciencias humanas. En este caso, el interés por la arqueología sería sobre todo un acto de sociabilidad. The author intends to show how the analysis of the social composition of the peruvian delegation at the first International Congress of Americanists, can offer a different view upon the anthropological movement in the XlXth century, as far as most of the delegates were more worried of their public image instead of the advancement of human sciences. In this particular case, their interest on archaeology would be seen as a social act.

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

    African Journals Online (AJOL)

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

  6. 21 CFR 864.7250 - Erythropoietin assay.

    Science.gov (United States)

    2010-04-01

    ... erythropoietin (an enzyme that regulates the production of red blood cells) in serum or urine. This assay provides diagnostic information for the evaluation of erythrocytosis (increased total red cell mass)...

  7. The comet assay: a heavenly method!

    Science.gov (United States)

    Collins, Andrew R

    2015-01-01

    The contributions to this special issue of Mutagenesis have been selected to cover the main research areas served by the comet assay, namely genotoxicology, environmental toxicology, human biomonitoring and fundamental investigations into mechanisms of DNA damage and repair. Innovative methods are described, technical issues are explored, and guidelines are given for venturing into relatively new or unexploited areas of research. The popularity of the comet assay in a historical context is illustrated by a bibliometric survey.

  8. Electrochemical Assay of Gold-Plating Solutions

    Science.gov (United States)

    Chiodo, R.

    1982-01-01

    Gold content of plating solution is assayed by simple method that required only ordinary electrochemical laboratory equipment and materials. Technique involves electrodeposition of gold from solution onto electrode, the weight gain of which is measured. Suitable fast assay methods are economically and practically necessary in electronics and decorative-plating industries. If gold content in plating bath is too low, poor plating may result, with consequent economic loss to user.

  9. Variables Affecting Two Electron Transport System Assays

    OpenAIRE

    Burton, G. Allen; Lanza, Guy R.

    1986-01-01

    Several methodological variables were critical in two commonly used electron transport activity assays. The dehydrogenase assay based on triphenyl formazan production exhibited a nonlinear relationship between formazan production (dehydrogenase activity) and sediment dilution, and linear formazan production occurred for 1 h in sediment slurries. Activity decreased with increased time of sediment storage at 4°C. Extraction efficiencies of formazan from sediment varied with alcohol type; methan...

  10. Evaluation of red blood cell Pig-a assay and PIGRET assay in rats using chlorambucil.

    Science.gov (United States)

    Maeda, Akihisa; Takahashi, Kei; Tsuchiyama, Hiromi; Oshida, Keiyu

    2016-11-15

    The Pig-a assay is a novel method to assess the in vivo mutagenicity of compounds, and it is expected to be useful for the detection of genotoxicity. In this study, to assess the performance of the Pig-a assay targeting red blood cells (RBCs; RBC Pig-a assay) and reticulocytes (RETs; PIGRET assay), chlorambucil, which is a genotoxicant, was orally administered to male rats once at 10, 20 and 40mg/kg on Day 1, and the mutant frequencies (MFs) of RBCs and RETs were examined periodically. In the RBC Pig-a assay, significant increases in MFs were observed at 40mg/kg on Day 15 and at 20mg/kg or higher on Day 29. In the PIGRET assay, MFs increased significantly at all dose levels on Day 8 and only at 20mg/kg on Day 15, but there was no increase in MFs in the treatment groups on Day 29. In conclusion, the RBC Pig-a assay and PIGRET assay in rats have sufficient sensitivity to detect the mutagenicity of chlorambucil, and the PIGRET assay could detect its mutagenicity earlier and at a lower dose than the RBC Pig-a assay. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Premier numéro bilingue

    Directory of Open Access Journals (Sweden)

    Esther Cloutier

    2008-05-01

    Full Text Available Nous sommes heureux de vous présenter ce nouveau numéro bilingue de la revue PISTES. Nous espérons ainsi faire connaître les travaux francophones sur la santé au travail dans le monde anglo-saxon. Nous vous rappelons que la traduction de ce numéro a été rendu possible grâce à une subvention du CRSH (Conseil de la recherche en sciences humaines du Canada ainsi qu’à la contribution de certains auteurs que nous tenons à remercier. Ce numéro aborde plusieurs thèmes de recherche reliés au travail...

  12. Film Premiere: Arrows of time at exploratorium

    CERN Document Server

    2007-01-01

    "Join award-wimming artist and filmmaker Ken McMullen for a uniqude presentation of films engaging ideas at the forefront of science and culture on Sunday, Apri 29 at 2pm at the Exploratorium." (1/3 page)

  13. Inkscape premiers pas en dessin vectoriel

    CERN Document Server

    Dufour, Nicolas

    2009-01-01

    Avec Inkscape, s'initier au dessin vectoriel devient un jeu d'enfant ! Paramétrez votre espace de travail pour créer avec aisance et précision. Intégrez des photos et des textes à vos dessins. Maniez les outils de forme ou dessinez à main levée avec les courbes de Bézier. Appliquez couleurs et dégradés aux formes et aux contours. Optimisez votre méthode de travail avec les modèles, les calques et les clones. Convertissez les objets en chemin et retouchez le détail de vos tracés. Transformez vos projets avec les filtres et les effets. Validez vos acquis avec 9 études de cas : création de logo, réalisation d'une invitation, conception d'une affiche...

  14. Sur les premiers peuplements du Pacifique sud

    OpenAIRE

    Semah, Anne-Marie; Detroit, F.

    2006-01-01

    About the first human settlements in South Pacific. South Pacific includes Australia and the Pacific Islands: Melanesia, Micronesia, Polynesia. Which human populations settled them? When, why and how? Where did they come from? Did environment influence these migrations? An exhaustive discussion of these vast questions could not be undertaken rigorously within the framework of this short article. We thus propose to discuss here very precise points that could bring a new lighting on the first s...

  15. Controlling variation in the comet assay

    Directory of Open Access Journals (Sweden)

    Andrew Richard Collins

    2014-10-01

    Full Text Available Variability of the comet assay is a serious issue, whether it occurs from experiment to experiment in the same laboratory, or between different laboratories analysing identical samples. Do we have to live with high variability, just because the comet assay is a biological assay rather than analytical chemistry? Numerous attempts have been made to limit variability by standardising the assay protocol, and the critical steps in the assay have been identified; agarose concentration, duration of alkaline incubation, and electrophoresis conditions (time, temperature and voltage gradient are particularly important. Even when these are controlled, variation seems to be inevitable. It is helpful to include in experiments reference standards, i.e. cells with a known amount of specific damage to the DNA. They can be aliquots frozen from a single large batch of cells, either untreated (negative controls or treated with, for example, H2O2 or X-rays to induce strand breaks (positive control for the basic assay, or photosensitiser plus light to oxidise guanine (positive control for Fpg- or OGG1-sensitive sites. Reference standards are especially valuable when performing a series of experiments over a long period - for example, analysing samples of white blood cells from a large human biomonitoring trial - to check that the assay is performing consistently, and to identify anomalous results necessitating a repeat experiment. The reference values of tail intensity can also be used to iron out small variations occurring from day to day. We present examples of the use of reference standards in human trials, both within one laboratory and between different laboratories, and describe procedures that can be used to control variation.

  16. Cell Culture Assay for Human Noroviruses [response

    Energy Technology Data Exchange (ETDEWEB)

    Straub, Tim M.; Honer Zu Bentrup, Kerstin; Orosz Coghlan, Patricia; Dohnalkova, Alice; Mayer, Brooke K.; Bartholomew, Rachel A.; Valdez, Catherine O.; Bruckner-Lea, Cindy J.; Gerba, Charles P.; Abbaszadegan, Morteza A.; Nickerson, Cheryl A.

    2007-07-01

    We appreciate the comments provided by Leung et al., in response to our recently published article “In Vitro Cell Culture Infectivity Assay for Human Noroviruses” by Straub et al. (1). The specific aim of our project was to develop an in vitro cell culture infectivity assay for human noroviruses (hNoV) to enhance risk assessments when they are detected in water supplies. Reverse transcription (RT) qualitative or quantitative PCR are the primary assays for waterborne NoV monitoring. However, these assays cannot distinguish between infectious vs. non-infectious virions. When hNoV is detected in water supplies, information provided by our infectivity assay will significantly improve risk assessment models and protect human health, regardless of whether we are propagating NoV. Indeed, in vitro cell culture infectivity assays for the waterborne pathogen Cryptosporidium parvum that supplement approved fluorescent microscopy assays, do not result in amplification of the environmentally resistant hard-walled oocysts (2). However, identification of life cycle stages in cell culture provides evidence of infectious oocysts in a water supply. Nonetheless, Leung et al.’s assertion regarding the suitability of our method for the in vitro propagation of high titers of NoV is valid for the medical research community. In this case, well-characterized challenge pools of virus would be useful for developing and testing diagnostics, therapeutics, and vaccines. As further validation of our published findings, we have now optimized RT quantitative PCR to assess the level of viral production in cell culture, where we are indeed finding significant increases in viral titer. The magnitude and time course of these increases is dependent on both virus strain and multiplicity of infection. We are currently preparing a manuscript that will discuss these findings in greater detail, and the implications this may have for creating viral challenge pools

  17. The Comet Assay: Tails of the (Unexpected. Use of the comet assay in pharmaceutical development.

    Directory of Open Access Journals (Sweden)

    Bas-jan Van Der Leede

    2015-08-01

    Full Text Available In genotoxicity testing of pharmaceuticals the rodent alkaline comet assay is being increasingly used as a second in vivo assay in addition to the in vivo micronucleus assay to mitigate in vitro positive results as recommended by regulatory guidance. In this presentation we want to give insight into the circumstances in vivo comet assay is deployed in a Genetic Toxicology Department of a pharmaceutical company. As the in vivo comet assay is a salvage assay, it means that some events have occurred in an in vitro assay and that the compound (or metabolite responsible for this signal is potentially deselected for further development. More than often the decision to perform an in vivo comet assay is at a very early stage in development and the first time that the compound will be tested in vivo at high/toxic dose levels. As almost no toxicokinetic data and tissue distribution data are available a careful design with maximizes the chances for successful mitigation is necessary. Decisions on acute or repeated dosing need to be made and arrangements for combining the in vivo comet assay with the in vivo micronucleus assay are to be considered. Often synthesis methods need to be scaled up fast to provide the required amount of compound and information on suitable formulations needs to be in place. As exposure data is crucial for interpretation of results, analytical methods need to be brought in place rapidly. An experienced multi skilled and communicative team needs to be available to deploy successfully this kind of assays at an early stage of development. We will present a few scenarios on study conduct and demonstrate how this assay can make a difference for the further development of a new drug.

  18. Random assay in radioimmunoassay: Feasibility and application compared with batch assay

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Jung Min; Lee, Hwan Hee; Park, Sohyun; Kim, Tae Sung; Kim, Seok Ki [Dept. of Nuclear MedicineNational Cancer Center, Goyang (Korea, Republic of)

    2016-12-15

    The batch assay has been conventionally used for radioimmunoassay (RIA) because of its technical robustness and practical convenience. However, it has limitations in terms of the relative lag of report time due to the necessity of multiple assays in a small number of samples compared with the random assay technique. In this study, we aimed to verify whether the random assay technique can be applied in RIA and is feasible in daily practice. The coefficients of variation (CVs) of eight standard curves within a single kit were calculated in a CA-125 immunoradiometric assay (IRMA) for the reference of the practically ideal CV of the CA-125 kit. Ten standard curves of 10 kits from 2 prospectively collected lots (pLot) and 85 standard curves of 85 kits from 3 retrospectively collected lots (Lot) were obtained. Additionally, the raw measurement data of both 170 control references and 1123 patients' sera were collected retrospectively between December 2015 and January 2016. A standard curve of the first kit of each lot was used as a master standard curve for a random assay. The CVs of inter-kits were analyzed in each lot, respectively. All raw measurements were normalized by decay and radioactivity. The CA-125 values from control samples and patients' sera were compared using the original batch assay and random assay. In standard curve analysis, the CVs of inter-kits in pLots and Lots were comparable to those within a single kit. The CVs from the random assay with normalization were similar to those from the batch assay in the control samples (CVs % of low/high concentration; Lot1 2.71/1.91, Lot2 2.35/1.83, Lot3 2.83/2.08 vs. Lot1 2.05/1.21, Lot2 1.66/1.48, Lot3 2.41/2.14). The ICCs between the batch assay and random assay using patients' sera were satisfactory (Lot1 1.00, Lot2 0.999, Lot3 1.00). The random assay technique could be successfully applied to the conventional CA-125 IRMA kits. The random assay showed strong agreement with the batch assay. The

  19. Using the CPTAC Assay Portal to identify and implement highly characterized targeted proteomics assays

    Science.gov (United States)

    Whiteaker, Jeffrey R; Halusa, Goran N; Hoofnagle, Andrew N; Sharma, Vagisha; MacLean, Brendan; Yan, Ping; Wrobel, John A; Kennedy, Jacob; Mani, DR; Zimmerman, Lisa J; Meyer, Matthew R.; Mesri, Mehdi; Abbatiello, Susan E; Boja, Emily; Carr, Steven A.; Chan, Daniel W.; Chen, Xian; Chen, Jing; Davies, Sherri R; Ellis, Matthew J. C.; Fenyö, David; Hiltke, Tara; Ketchum, Karen A.; Kinsinger, Chris; Kuhn, Eric; Liebler, Daniel C.; Lin, De; Liu, Tao; Loss, Michael; MacCoss, Michael J; Qian, Wei-Jun; Rivers, Robert; Rodland, Karin D.; Ruggles, Kelly V; Scott, Mitchell G; Smith, Richard D.; Thomas, Stefani; Townsend, R. Reid; Whiteley, Gordon; Wu, Chaochao; Zhang, Hui; Zhang, Zhen; Rodriguez, Henry; Paulovich, Amanda G

    2016-01-01

    Summary The Clinical Proteomic Tumor Analysis Consortium (CPTAC) of the National Cancer Institute (NCI) has launched an Assay Portal (http://assays.cancer.gov) to serve as an open-source repository of well-characterized targeted proteomic assays. The portal is designed to curate and disseminate highly characterized, targeted mass spectrometry (MS)-based assays by providing detailed assay performance characterization data, standard operating procedures, and access to reagents. Assay content is accessed via the portal through queries to find assays targeting proteins associated with specific cellular pathways, protein complexes, or specific chromosomal regions. The position of the peptide analytes for which there are available assays are mapped relative to other features of interest in the protein, such as sequence domains, isoforms, single nucleotide polymorphisms, and post-translational modifications. The overarching goals are to enable robust quantification of all human proteins and to standardize the quantification of targeted MS-based assays to ultimately enable harmonization of results over time and across laboratories. PMID:26867747

  20. Using the CPTAC Assay Portal to identify and implement highly characterized targeted proteomics assays

    Energy Technology Data Exchange (ETDEWEB)

    Whiteaker, Jeffrey R.; Halusa, Goran; Hoofnagle, Andrew N.; Sharma, Vagisha; MacLean, Brendan; Yan, Ping; Wrobel, John; Kennedy, Jacob; Mani, DR; Zimmerman, Lisa J.; Meyer, Matthew R.; Mesri, Mehdi; Boja, Emily; Carr, Steven A.; Chan, Daniel W.; Chen, Xian; Chen, Jing; Davies, Sherri; Ellis, Matthew; Fenyo, David; Hiltket, Tara; Ketchum, Karen; Kinsinger, Christopher; Kuhn, Eric; Liebler, Daniel; Liu, Tao; Loss, Michael; MacCoss, Michael; Qian, Weijun; Rivers, Robert; Rodland, Karin D.; Ruggles, Kelly; Scott, Mitchell; Smith, Richard D.; Thomas, Stefani N.; Townsend, Reid; Whiteley, Gordon; Wu, Chaochao; Zhang, Hui; Zhang, Zhen; Rodriguez, Henry; Paulovich, Amanda G.

    2016-02-12

    The Clinical Proteomic Tumor Analysis Consortium (CPTAC) of the National Cancer Institute (NCI) has launched an Assay Portal (http://assays.cancer.gov) to serve as an open-source repository of well-characterized targeted proteomic assays. The portal is designed to curate and disseminate highly characterized, targeted mass spectrometry (MS)-based assays by providing detailed assay performance characterization data, standard operating procedures, and access to reagents. Assay content is accessed via the portal through queries to find assays targeting proteins associated with specific cellular pathways, protein complexes, or specific chromosomal regions. The position of the peptide analytes for which there are available assays are mapped relative to other features of interest in the protein, such as sequence domains, isoforms, single nucleotide polymorphisms, and post-translational modifications. The overarching goals are to enable robust quantification of all human proteins and to standardize the quantification of targeted MS-based assays to ultimately enable harmonization of results over time and across laboratories.

  1. Comet assay on mice testicular cells

    Directory of Open Access Journals (Sweden)

    Anoop Kumar Sharma

    2015-05-01

    Full Text Available Heritable mutations may result in a variety of adverse outcomes including genetic disease in the offspring. In recent years the focus on germ cell mutagenicity has increased and the “Globally Harmonized System of Classification and Labelling of Chemicals (GHS” has published classification criteria for germ cell mutagens (Speit et al., 2009. The in vivo Comet assay is considered a useful tool for investigating germ cell genotoxicity. In the present study DNA strand breaks in testicular cells of mice were investigated. Different classes of chemicals were tested in order to evaluate the sensitivity of the comet assay in testicular cells. The chemicals included environmentally relevant substances such as Bisphenol A, PFOS and Tetrabrombisphenol A. Statistical power calculations will be presented to aid in the design of future Comet assay studies on testicular cells. Power curves were provided with different fold changes in % tail DNA, different number of cells scored and different number of gels (Hansen et al., 2014. An example is shown in Figure 1. A high throughput version of the Comet assay was used. Samples were scored with a fully automatic comet assay scoring system that provided faster scoring of randomly selected cells.

  2. Research highlights: digital assays on chip.

    Science.gov (United States)

    Kim, Donghyuk; Wei, Qingshan; Kong, Janay Elise; Ozcan, Aydogan; Di Carlo, Dino

    2015-01-07

    The ability to break up a volume of fluid into smaller pieces that are confined or separated to prevent molecular communication/transport is a key capability intrinsic to microfluidic systems. This capability has been used to develop or implement digital versions of traditional molecular analysis assays, including digital PCR and digital immunoassays/ELISA. In these digital versions, the concentration of the target analyte is in a range such that, when sampled into smaller fluid volumes, either a single molecule or no molecule may be present. Subsequent amplification is sensitive enough to obtain a digital readout of the presence of these target molecules. Advantages of such approaches that are claimed include quantification without calibration and robustness to variations in reaction conditions or times because the digital readout is less sensitive to absolute signal intensity levels. Weaknesses of digital approaches include a lower dynamic range of concentrations over which the assay is sensitive, which depends on the total volume that can be analyzed. We highlight recent efforts to expand the dynamic range of digital assays based on exploiting reaction/diffusion phenomena. A side-by-side study that evaluates the strengths of digital assays reveals that the majority of these claims are supported, with specific caveats. Finally, we highlight approaches to apply digital assays to analyze new types of reactions, including the active transport of protons across membranes by ATPases at the single protein level - perhaps opening up new biophysical understanding and screening opportunities, similar to widely deployed single-molecule ion channel analysis.

  3. Théorie visco-élastique non-extensive IV. Premiers tests expérimentaux de la théorie simplifiée à modes translationnels

    Science.gov (United States)

    Volino, F.

    This paper describes the first tests of the non-extensive visco-elastic theory with translational modes developed in (III), using viscosity, self-diffusion, shear elasticity, thermal conductivity and surface tension literature data of several liquids, namely water, organic solvents and a nematics, for which numerous data exist in the literature. As in the rotational case (I, II), a remarkable qualitative and quantitative agreement is obtained. The influence of translational-rotational coupling on the flow is examined for the nematics and for water. The obtained results, of equivalent importance as those obtained previouly in (II), are more spectacular in the sense that they concern more common substances. If the proposed description is accepted, important changes in the meaning of some physical concepts are necessarily implied. Cet article décrit les premiers tests de la théorie visco-élastique non-extensive à modes translationnels développée dans l'article (III), à l'aide de données de viscosité, de coefficients d'auto-diffusion, d'élasticité de cisaillement, de conductivité thermique et de tension superficielle, d'un certain nombre de liquides, à savoir l'eau, des solvants organiques usuels et un nématique, pour lesquels de très nombreuses données expérimentales existent dans la littérature. Comme dans le cas rotationnel avec les nématiques (I, II), un accord qualitatif et surtout quantitatif remarquable est obtenu. L'influence du couplage translation-orientation sur les propriétés d'écoulement est examiné dans le cas du nématique, et dans une certaine mesure, de l'eau. Ces résultats, de même importance absolue que ceux obtenus dans (II) avec les aspects rotationnels, sont relativement plus spectaculaires dans la mesure où ils sont relatifs à des substances beaucoup plus communes. Si ce type de description est accepté, ils impliquent inévitablement des changements importants sur la signification de certains concepts physiques.

  4. La stigmatisation particulière du pédéraste passif dans les enquêtes de médecine légale dans la premiere partie du xixe siècle

    Directory of Open Access Journals (Sweden)

    Thierry Pastorello

    2010-01-01

    Full Text Available Les enquêtes de médecine légale dans la première partie du xixe siècle sont les premiers travaux envisageant la pratique de l’homosexualité masculine sous l’angle scientifique. Deux figures sont primordiales : le docteur Pierre-Jean-Georges Cabanis (1757-1808 membre des idéologues et qui entendait faire de la médecine la pièce maîtresse d’une science de l’homme et Ambroise Tardieu 1818-1879 professeur de médecine légale à Paris auteur d’un traité vu comme un aboutissement de ces études médico-légales. Il faut signaler l’influence de la phrénologie. Ces médecins définissent des stigmates physiques et moraux propres à la pratique de la pédérastie, surtout passive et à l’occasion définissent un personnage type. Ces discours relèvent d’abord d’une phraséologie morale et ils puisent leurs racines antérieurement. Cependant il faut aussi replacer ces discours stigmatisant la pédérastie passive dans le contexte de la construction de la virilité moderne. Il faut aussi placer la médecine légale dans le contexte de la médecine et de ses évolutions.The particular stigmatization of the passive pederast in the inquiries of forensic medecine in the first part of the XIX centuryForensic medicine in the first part of the 19th century investigations are first work considering the practice of male homosexuality in scientific terms. Two figures are crucial: the Dr. Pierre-Jean-Georges Cabanis (1757-1808 the ideologists member and intended to do medicine the centrepiece of a human science and Ambroise Tardieu 1818 - 1879 Professor of forensic medicine in Paris author of a treaty as a culmination of these forensic studies. It should be noted the influence of the phrenology. These physicians define physical stigma and moral own practice of pederasty, especially passive and during the define a type character. These speeches are firstly a moral phraseology and they draw their roots previously. However we also

  5. EDTA interference in electrochemiluminescence ACTH assay.

    Science.gov (United States)

    Toprak, Burak; Yalcin, Hulya; Arı, Elif; Colak, Ayfer

    2016-11-01

    Background As plasma is the recommended sample type for Roche adrenocorticotropic hormone (ACTH) assay, we evaluated the effect of EDTA concentration on Cobas ACTH assay. Methods Samples containing twofold and fourfold higher concentrations of EDTA were prepared by adding plasma to empty K2EDTA tubes and by making under-filled EDTA tubes. All measurements were performed with four replicates. Results Increased EDTA concentration resulted in a significant decrease in ACTH concentration. Fifty-per cent-filled EDTA tube showed 19% decrease in ACTH concentration and 25% filled EDTA tube showed 50% decrease in ACTH concentration. Conclusion We recommend that inadequately filled EDTA specimens should be rejected when using Cobas ACTH assay.

  6. Comet assay on tetraploid yeast cells

    DEFF Research Database (Denmark)

    Rank, Jette; Syberg, Kristian; Jensen, Klara

    2009-01-01

    Tetraploid yeast cells (Saccharomyces cerevisiae) were used in the comet assay with the intention of developing a new, fast and easy assay for detecting environmental genotoxic agents without using higher organisms. Two DNA-damaging chemicals, H2O2 and acrylamide, together with wastewater from...... three municipal treatment plants were tested for their effect on the yeast-cell DNA. The main problem with using yeast in the comet assay is the necessity to degrade the cell wall. This was achieved by using Zymolase 100 T twice during the procedure, since Zymolase 20 T did not open the cell wall....... Analytical problems that arose due to the small amount of DNA in the yeast nuclei in haploid and diploid cells, which contain 13 Mbp and 26 Mbp DNA per cell, respectively, were solved by using tetraploid yeast cells (52 Mbp) instead. DNA damage was shown after exposure to H2O2 and acrylamide. The lowest dose...

  7. Fungicide resistance assays for fungal plant pathogens.

    Science.gov (United States)

    Secor, Gary A; Rivera, Viviana V

    2012-01-01

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

  8. Detection of Eperythrozoon wenyoni by PCR assay

    Institute of Scientific and Technical Information of China (English)

    Jian WANG; Yutao ZHU; Jianhua QIN; Fumei ZHANG; Yuelan ZHAO

    2009-01-01

    The objective of this research was to develop a detection method for Eperythrozoon wenyoni infection using polymerase chain reaction (PCR) assay technique. A pair of primers was designed and synthesized according to the conservative sequence 16S rRNA. The PCR assay was performed with the primers. A 985-bp fragment was amplified by using PCR. The amplified fragments with the expected size were identified by EcoR I restriction digestion. The crossing-reaction, specific-reaction and duplicate-reaction indicated that the PCR method is a specific, sensitive, fast and effective method for diagnosing E. Wenyoni infection at group level.

  9. Instructions for Uploading Data to the Assay Portal - Instructions for Uploading Data to the Assay Portal

    Science.gov (United States)

    This document provides instructions for configuring and uploading data files to the CPTAC Assay Portal. It is divided into sections, with an overview checklist provided at the end. If help is needed at any stage of the process, please use the support page: https://assays.cancer.gov/support/

  10. Development and Application of High-flux Molecular Differential Detection System with LAMP Assay for 10 Foodborne Pathogen%10种食源性疾病病原体高通量LAMP分子鉴别检测体系的建立及应用

    Institute of Scientific and Technical Information of China (English)

    孙边成; 长如胜; 宋克云; 欧新华; 姚栋; 苏良; 叶文; 陈法明

    2011-01-01

    viruses pathogens (Norwalk virus and Rotavirus). Each of 25 kinds of common intestinal bacteria and viruses was amplified in a water bath at 65°C for 60 min (LAMP) or 120 min (RT- LAMP), the results were judged by electrophoresis and the naked eye after completion of amplification. 10 kinds of LAMP methods for foodborne pathogens were applied for detection of 365 specimens of vomit, anal swabs, food samples and other specimens from foodborne disease. Results Ten kinds of LAMP or RT - LAMP methods for foodborne pathogens only amplified the target pathogens, the results showed the LAMP - specific laddering with electro-phoretic analysis and the LAMP reaction mixture turned green judgment by naked eye after the addition of SYBR Green I. The : sensitivities of 8 kinds of LAMP for bacterial pathogens were 3 x 10° ~ 1.2 x 102 cfu/ml, 6 kinds of LAMP for bacterial pathogens were higher than that of PCR assay, and the sensitivities of the other pathogens (Vibrio cholerae, EHEC O157-.H7, Norwalk virus and Rotavirus) were equal to those of PCR assay. In 365 specimens from foodborne disease, there were 11 samples of Salmonella, 6 samples of Shigella, 13 sanples of S. Aureus, 2 samples of EHEC 0157: H7, 2 samples of V. Parahaemolyticus, 2 samples of L. Monocytogenes, 12 samples of Norwalk virus and 7 samples of Rotavirus detected with 10 kinds of LAMP methods for foodborne pathogens. Conclusions This study established a relatively complete molecular differential detection system with LAMP, which could be applied to simultaneous high - flux detection of 10 kinds of foodborne pathogens in a water bath, moreover, the detection system had the advantages of rapidity, without needing special instru-merits, and judgment with the naked eye based on the color of reaction mixture. It could be potentially applied to detection of DNA of foodborne pathogens in poor - equipped laboratories.

  11. Drugs and brain death: drug assay perspectives.

    Science.gov (United States)

    Morris, R G

    1996-08-01

    The ability to make any meaningful interpretation of a drug assay result is very dependent upon a knowledge of the limitations of the method(s) used (sensitivity, specificity etc.), and the concentration that may be measured in plasma and its relationship to CNS effects. We need more information about 'critical' concentrations for each drug and sedation in the setting of the brain-injured patient before meaningful interpretation can be applied to such data. While the above discussion is critical of screen-type assays, the alternative specific assays are not easily provided for, as obviously the resourcing of laboratories to be able to deliver such specialized services for a range of therapeutic drugs, in addition to 'social' drugs or other toxins (e.g. glues, pesticides, solvents, environmental substances etc), becomes an increasingly complex issue in the current economic climate. Hence, the analytical laboratory can offer valuable support to the clinical team however, the interpretation of such results must be assessed in the light of many limitations of such assay methods and not seen as the 'gold standard' for assessment of brain function.

  12. Functionalized Nanofiber Meshes Enhance Immunosorbent Assays.

    Science.gov (United States)

    Hersey, Joseph S; Meller, Amit; Grinstaff, Mark W

    2015-12-01

    Three-dimensional substrates with high surface-to-volume ratios and subsequently large protein binding capacities are of interest for advanced immunosorbent assays utilizing integrated microfluidics and nanosensing elements. A library of bioactive and antifouling electrospun nanofiber substrates, which are composed of high-molecular-weight poly(oxanorbornene) derivatives, is described. Specifically, a set of copolymers are synthesized from three 7-oxanorbornene monomers to create a set of water insoluble copolymers with both biotin (bioactive) and triethylene glycol (TEG) (antifouling) functionality. Porous three-dimensional nanofiber meshes are electrospun from these copolymers with the ability to specifically bind streptavidin while minimizing the nonspecific binding of other proteins. Fluorescently labeled streptavidin is used to quantify the streptavidin binding capacity of each mesh type through confocal microscopy. A simplified enzyme-linked immunosorbent assay (ELISA) is presented to assess the protein binding capabilities and detection limits of these nanofiber meshes under both static conditions (26 h) and flow conditions (1 h) for a model target protein (i.e., mouse IgG) using a horseradish peroxidase (HRP) colorimetric assay. Bioactive and antifouling nanofiber meshes outperform traditional streptavidin-coated polystyrene plates under flow, validating their use in future advanced immunosorbent assays and their compatibility with microfluidic-based biosensors.

  13. Endoproteolytic activity assay in malting barley

    Directory of Open Access Journals (Sweden)

    Blanca Gómez Guerrero

    2013-12-01

    Full Text Available Hydrolysis of barley proteins into peptides and amino acids is one of the most important processes during barley germination.The degradation of the endosperm stored proteins facilitates water and enzyme movements, enhances modification, liberates starch granules and increases soluble amino nitrogen. Protease activity is the result of the activities of a mixture of exo- and endo-proteases. The barley proteins are initially solubilized by endo-proteases and the further by exo-proteases. Four classes of endo-proteases have been described: serine-proteases, cysteine-proteases, aspartic-proteases and metallo-proteases. The objective of this work was to develop a rapid and colorimetric enzymatic assay to determine the endo-proteolytic activity of the four endo-protease classes using two different substrates: azo-gelatin and azo-casein. Optimum conditions for the assays such as: pH,reaction time and temperature and absorbance scale were determined. Azo-gelatin presented several difficulties in standardizing an “in solution” assay. On the other hand, azo-casein allowed standardization of the assay for the four enzyme classes to produce consistent results. The endo-proteoteolytic method developed was applied to determine the endo-protease activity in barley, malt and wort.

  14. Comet assay on mice testicular cells

    DEFF Research Database (Denmark)

    Sharma, Anoop Kumar

    2015-01-01

    for germ cell mutagens (Speit et al., 2009). The in vivo Comet assay is considered a useful tool for investigating germ cell genotoxicity. In the present study DNA strand breaks in testicular cells of mice were investigated. Different classes of chemicals were tested in order to evaluate the sensitivity...

  15. In vitro solubility assays in drug discovery.

    Science.gov (United States)

    Kerns, Edward H; Di, Li; Carter, Guy T

    2008-11-01

    The solubility of a compound depends on its structure and solution conditions. Structure determines the lipophilicity, hydrogen bonding, molecular volume, crystal energy and ionizability, which determine solubility. Solution conditions are affected by pH, co-solvents, additives, ionic strength, time and temperature. Many drug discovery experiments are conducted under "kinetic" solubility conditions. In drug discovery, solubility has a major impact on bioassays, formulation for in vivo dosing, and intestinal absorption. A good goal for the solubility of drug discovery compounds is >60 ug/mL. Equilibrium solubility assays can be conducted in moderate throughput, by incubating excess solid with buffer and agitating for several days, prior to filtration and HPLC quantitation. Kinetic solubility assays are performed in high throughput with shorter incubation times and high throughput analyses using plate readers. The most frequently used of these are the nephelometric assay and direct UV assay, which begin by adding a small volume of DMSO stock solution of each test compound to buffer. In nephelometry, this solution is serially diluted across a microtitre plate and undissolved particles are detected via light scattering. In direct UV, undissolved particles are separated by filtration, after which the dissolved material is quantitated using UV absorption. Equilibrium solubility is useful for preformulation. Kinetic solubility is useful for rapid compound assessment, guiding optimization via structure modification, and diagnosing bioassays. It is often useful to customize solubility experiments using conditions that answer specific research questions of drug discovery teams, such as compound selection and vehicle development for pharmacology and PK studies.

  16. Benzodiazepine Synthesis and Rapid Toxicity Assay

    Science.gov (United States)

    Fletcher, James T.; Boriraj, Grit

    2010-01-01

    A second-year organic chemistry laboratory experiment to introduce students to general concepts of medicinal chemistry is described. Within a single three-hour time window, students experience the synthesis of a biologically active small molecule and the assaying of its biological toxicity. Benzodiazepine rings are commonly found in antidepressant…

  17. Production and assay of forskolin antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Ho, L.T.; Ho, R.J.

    1986-05-01

    Forskolin (Fo), a cardiovascular active diterpene of plant origin, has been widely used as a research tool in regulation of the catalytic activity of adenylate cyclase (AC). A linear relationship of Fo binding to plasma membrane with activation of AC has been reported. The present abstract describes the production and assay of Fo antibodies (AB). 7-0-Hemisuccinyl-7-deacetyl Fo, coupled to either human serum albumin or goat IgG, was injected into goats to elicit AB to Fo haptan. AB to Fo in antiserum or an isolated IgG fraction was tested by two assay methods, a radioimmunoassay using /sup 3/H-Fo as a tracer and a colorimetric enzyme-linked immunosorbent assay (ELISA) using horse radish peroxidase-rabbit anti goat IgG as indicator. The titers for Fo antiserum were 4000-10,000. In the defined assay condition, approximately 20-25% of the added /sup 3/H-Fo was found to bind to AB. The bound radioactivity was displaced by Fo-HSA or Fo-goat IgG or free unlabelled Fo ranging from 0.5-50 pmol/tube, or 5-500 nM. The IC/sub 50/ was approximately 8-10 pmol/tube or 80-100 nM. The binding of HRP-rabbit anti goat IgG in the ELISA was inhibited by proper Fo conjugate. The development of methods for production and assay for Fo AB may be useful in the study of mechanism of activation of AC by Fo and Fo-like compound.

  18. Development of an Easy and High-Throughput Cell Assay System with a Culture Chip and an Assay Chip

    Science.gov (United States)

    Sugiura, Kanako; Kaji, Noritada; Okamoto, Yukihiro; Tokeshi, Manabu; Baba, Yoshinobu

    High throughput cell assay is significantly important in drug screening, assessment of toxicity etc. Cell assay with a microchip is one of the candidates for high throughput cell assay. However, reported cell assay system with the microchip requires expensive apparatus for refluxing medium and investigation of optimum experimental condition for steady data. For an inexpensive, easy and high throughput cell assay, we introduce a new cell assay system combined with a culture chip and an assay chip made of poly(dimethyl siloxane). Cell culture chips enabled cell to proliferate along the microchannel without refluxing medium and permitted to prepare cell patterning easily. Also, assay chips formed concentration gradient inside the chip and allowed the cell assay with different concentrations of drug at the same time. Thus, our developed cell assay system can overcome the problems of the present cell assay and would promote the drug discovery, assessment of toxicity etc.

  19. Tadmekka. Archéologie d’une ville caravanière des premiers temps du commerce transsaharien Tadmekka: The archaeology of an early cross-Saharan caravan town in West Africa

    Directory of Open Access Journals (Sweden)

    Sam Nixon

    2013-05-01

    Full Text Available Cet article présente les premières fouilles systématiques de Tadmekka, une des villes commerçantes majeures d’Afrique de l’Ouest, qui a participé à la croissance du commerce transsaharien durant les premières années de l’ère islamique (vers 650-1500. En 2005, des fouilles sur les ruines de Tadmekka (Essouk, dans le Nord du Mali, ont fourni une stratigraphie de 6,5 m, depuis le milieu du premier millénaire jusque vers 1400. Les analyses effectuées ensuite ont considérablement amélioré notre compréhension des périodes « préhistoriques » et historiques, et permettent d’ouvrir des perspectives nouvelles. Après une description des prémisses du projet et une présentation complète de ses résultats, nous présentons une histoire du développement de Tadmekka, en plaçant côte à côte les résultats archéologiques et les indices fournis par la documentation historique. Nous nous focalisons ensuite sur des analyses spécifiques, comme par exemple : les modes de construction à Tadmekka, le monnayage en or, le commerce des produits au-delà du Sahara, les céramiques, les industries locales et les pratiques alimentaires.This paper reports the first systematic excavations of Tadmekka, one of the major West African trading towns that enabled the huge growth of cross-Saharan trade during the early Islamic era (c. AD 650-1500. In 2005 excavations at Tadmekka’s contemporary site (‘Essouk’ in northern Mali yielded a 6.5 m excavated sequence, dating from the mid-first millennium AD to c. 1400. Subsequent analysis has significantly improved understanding of the town’s ‘prehistoric’ and historic periods, and shown how the archaeology presents a wealth of evidence of significance for wider debates. Following a background to the project and a full presentation of its results, we build up an improved story of Tadmekka’s development, by placing the archaeology side by side with the evidence provided by early

  20. Le diagnostic anténatal de la trisomie 21 par l'hybridation in situ en fluorescence (FISH): à propos des premiers tests réalisés au Maroc

    Science.gov (United States)

    Lamzouri, Afaf; Natiq, Abdelhafid; Tajir, Mariam; Sendid, Mohamed; Sefiani, Abdelaziz

    2012-01-01

    Introduction Le but de cette étude était de présenter les premiers résultats de diagnostic anténatal de la trisomie 21 par la technique d'hybridation in situ en fluorescence (FISH) au Maroc et discuter son intérêt dans le diagnostic rapide de cette aneuploïdie. Méthodes Ce travail a été réalisé chez 23 femmes avec des grossesses à haut risque de trisomie 21. La moyenne d’âge des gestantes étaient de 37,43 ans avec des extrêmes de 21 et 43 ans. Toutes étaient musulmanes mariées, mariage légitimé par la Charia, dont trois mariages consanguins, sauf une originaire de la République Démocratique du Congo qui était chrétienne et concubine. La majorité des femmes étaient fonctionnaires et avaient un niveau de scolarisation moyen à élevé. Toutes les patientes ont bénéficié d'une consultation de génétique médicale au cours de laquelle il leur a été donné des informations sur la technique, son intérêt et ses limites. Il s'agit de femmes enceintes qui avaient soit un âge maternel élevé ou des signes d'appel échographiques et/ ou biochimiques. Une des patientes était porteuse d'une translocation robertsonienne t(14;21) équilibrée. Une amniocentèse a été réalisée chez toutes les gestantes et aucun avortement n'a était induit par ce geste invasif. L’âge gestationnel moyen à la première consultation était de 14 semaines d'aménorrhée (SA) et à l'amniocentèse était de 16 SA et 5 jours. L'analyse FISH a été réalisée, après consentement des couples, sur des cellules non cultivées à partir des échantillons de liquides amniotiques, en utilisant des sondes spécifiques du chromosome 21. Résultats Parmi les 23 patientes qui ont bénéficiées d'un diagnostic anténatal de la trisomie 21 par la technique FISH, nous avons pu rassurer 21 d'entre elles, et nous avons détecté deux cas de trisomie 21 fœtal. Conclusion La technique FISH permet un diagnostic anténatal rapide, en moins de 48h, de la trisomie 21 sur

  1. Lump corrections for radioactive waste assay.

    Science.gov (United States)

    Miller, T J

    2009-09-01

    Previous studies have shown that automated radioactive waste assay techniques, such as segmented gamma scanner (SGS) and automated qualitative and quantitative (AQ2), have severely underestimated fissile material due to either the malfunction or absence of appropriate lump correction routines. This paper examines the application of manual techniques, such as Monte Carlo N particle (MCNP) and spectral non-destructive assay platform (SNAP) software, to lump corrections in plutonium (Pu), enriched uranium (EU) and depleted uranium (DU) waste streams. Excellent results have been obtained when comparing MCNP with SNAP and applying the SNAP lump correction routine to a range of simulated and typical wastes containing various Pu and EU lump sizes. It has been concluded that the need for lump corrections was relatively rare and usually apparent from abnormal gamma ray peak area ratios, since most AWE waste streams are only lightly shielded.

  2. Transient expression assays in tobacco protoplasts.

    Science.gov (United States)

    Vanden Bossche, Robin; Demedts, Brecht; Vanderhaeghen, Rudy; Goossens, Alain

    2013-01-01

    The sequence information generated through genome and transcriptome analysis from plant tissues has reached unprecedented sizes. Sequence homology-based annotations may provide hints for the possible function and roles of particular plant genes, but the functional annotation remains nonexistent or incomplete for many of them. To discover gene functions, transient expression assays are a valuable tool because they can be done more rapidly and at a higher scale than generating stably transformed tissues. Here, we describe a transient expression assay in protoplasts derived from suspension cells of tobacco (Nicotiana tabacum) for the study of the transactivation capacities of transcription factors. To enhance throughput and reproducibility, this method can be automated, allowing medium-throughput screening of interactions between large compendia of potential transcription factors and gene promoters.

  3. Identification of irradiated pepper with comet assay

    Energy Technology Data Exchange (ETDEWEB)

    Prieto Miranda, Enrique Fco.; Moreno Alvarez, Damaris L.; Carro Palacio, Sandra [Centro de Aplicaciones Tecnologicas y Desarrollo Nuclear. (CEADEN), Ciudad de La Habana (Cuba)]. E-mail: efprieto@ceaden.edu.cu; damaris@ceaden.edu.cu; Iglesia Enriquez, Isora [Instituto de Investigacion para la Industria Alimenticia (IIIA), Ciudad de La Habana (Cuba)

    2007-07-01

    The treatment of foods with ionizing radiations is a technological process utilized in order to increase the hygienic quality and the storage time of the foods. Several methods of detection of irradiated foods have been recommended. The comet assay of DNA is one fast and economical technique for the qualitative identification of irradiated foods. The objective of the present paper was to identify with the comet assay technique the modifications of the DNA molecule of irradiated pepper storage at environment and refrigeration temperatures and different post-irradiation times for different absorbed dose values, (0.1, 0.3 and 0.5 kGy). It was demonstrated that for the high absorbed dose values was observed a greater break into fragments of the DNA molecule, which shows the application of this technique for the identification of irradiated foods. (author)

  4. Posttranslational Modification Assays on Functional Protein Microarrays.

    Science.gov (United States)

    Neiswinger, Johnathan; Uzoma, Ijeoma; Cox, Eric; Rho, HeeSool; Jeong, Jun Seop; Zhu, Heng

    2016-10-03

    Protein microarray technology provides a straightforward yet powerful strategy for identifying substrates of posttranslational modifications (PTMs) and studying the specificity of the enzymes that catalyze these reactions. Protein microarray assays can be designed for individual enzymes or a mixture to establish connections between enzymes and substrates. Assays for four well-known PTMs-phosphorylation, acetylation, ubiquitylation, and SUMOylation-have been developed and are described here for use on functional protein microarrays. Phosphorylation and acetylation require a single enzyme and are easily adapted for use on an array. The ubiquitylation and SUMOylation cascades are very similar, and the combination of the E1, E2, and E3 enzymes plus ubiquitin or SUMO protein and ATP is sufficient for in vitro modification of many substrates.

  5. Comparative assay of Vipera ammodytes antivenom potency.

    Science.gov (United States)

    Capitanescu, Cristian; Macovei Oprescu, Anca Monica; Supeanu, Alexandru; Coculescu, Bogdan Ioan; Strambu, Victor; Macovei, Radu Alexandru; Manole, Gheorghe

    2016-12-01

    The finding of the most appropriate way to assess precisely the antivenom efficacy represents one of the major issues for antivenom standardization and success increasing of antivenom therapy. The efficacy of experimental Vipera ammodytes antivenom raised in sheep was determined using in vivo mouse lethality test, respectively, L-aminoacid oxidase, total proteinase and phospholipase A2 antienzymatic effectiveness. The values gained for the antivenom potency depend on the method of measure. So, some of the most toxic venom proteins own phospholipase A2 activity and provide the highest antivenom potency (lowest effective dose) values by antienzymatic assay method. This value is similar with total antiproteolytic antivenom potency value, but almost three times higher than value obtained by L-aminoacid oxidase (low toxic viper venom protein) antienzymatic assay method.

  6. Kinetic viability assays using DRAQ7 probe.

    Science.gov (United States)

    Wlodkowic, Donald; Akagi, Jin; Dobrucki, Jurek; Errington, Rachel; Smith, Paul J; Takeda, Kazuo; Darzynkiewicz, Zbigniew

    2013-07-01

    Cell death within cell populations is a stochastic process where cell-to-cell variation in temporal progression through the various stages of cell death arises from asynchrony of subtle fluctuations in the signaling pathways. Most cell death assays rely on detection of the specific marker of cell demise at the end-point of cell culturing. Such an approach cannot account for the asynchrony and the stochastic nature of cell response to the death-inducing signal. There is a need therefore for rapid and high-throughput bioassays capable of continuously tracking viability of individual cells from the time of encountering a stress signal up to final stages of their demise. In this context, a new anthracycline derivative, DRAQ7, is gaining increasing interest as an easy-to-use marker capable of long-term monitoring of cell death in real-time. This novel probe neither penetrates the plasma membrane of living cells nor does it affect the cells' susceptibility to the death-inducing agents. However, when the membrane integrity is compromised, DRAQ7 enters cells undergoing demise and binds readily to nuclear DNA to report cell death. Here, we provide three sets of protocols for viability assays using DRAQ7 probe. The first protocol describes the innovative use of single-color DRAQ7 real-time assay to dynamically track cell viability. The second protocol outlines a simplified end-point DRAQ7 staining approach. The final protocol highlights the real-time and multiparametric apoptosis assay utilizing DRAQ7 dye concurrently with tetramethylrhodamine methyl ester (TMRM), the mitochondrial trans-membrane electrochemical potential (ΔΨm) sensing probe.

  7. A new fluorescent assay for sialyltransferase.

    Science.gov (United States)

    Kajihara, Y; Kamitani, T; Sakakibara, T

    2001-04-23

    A new fluorescent assay for the sialyltransferase reaction was established. After incubation of the sialyltransferase reaction, the sialyloligosaccharide obtained was treated by acid hydrolysis, and then the NeuAc that was released was labeled with 1,2-diamino-4,5-methylenedioxibenzene. The fluorescent-labeled NeuAc could be estimated by HPLC (excitation: 373 nm; emission: 448 nm) and a Lineweaver-Burk plot could be plotted with the data from this analysis.

  8. Delivery of High-Quality Biomarker Assays

    Directory of Open Access Journals (Sweden)

    Brian N. Swanson

    2002-01-01

    Full Text Available Biomarker measurements now support key decisions throughout the drug development process, from lead optimization to regulatory approvals. They are essential for documenting exposure-response relationships, specificity and potency toward the molecular target, untoward effects, and therapeutic applications. In a broader sense, biomarkers constitute the basis of clinical pathology and laboratory medicine. The utility of biomarkers is limited by their specificity and sensitivity toward the drug or disease process and by their overall variability. Understanding and controlling sources of variability is not only imperative for delivering high-quality assay results, but ultimately for controlling the size and expense of research studies. Variability in biomarker measurements is affected by: biological and environmental factors (e.g., gender, age, posture, diet and biorhythms, sample collection factors (e.g., preservatives, transport and storage conditions, and collection technique, and analytical factors (e.g., purity of reference material, pipetting precision, and antibody specificity. The quality standards for biomarker assays used in support of nonclinical safety studies fall under GLP (FDA regulations, whereas, those assays used to support human diagnostics and healthcare are established by CLIA (CMS regulations and accrediting organizations such as the College of American Pathologists. While most research applications of biomarkers are not regulated, biomarker laboratories in all settings are adopting similar laboratory practices in order to deliver high-quality data. Because of the escalation in demand for biomarker measurements, the highly-parallel (multi-plexed assay platforms that have fueled the rise of genomics will likely evolve into the analytical engines that drive the biomarker laboratories of tomorrow.

  9. Methods and devices for protein assays

    Science.gov (United States)

    Chhabra, Swapnil; Cintron, Jose M.; Shediac, Renee

    2009-11-03

    Methods and devices for protein assays based on Edman degradation in microfluidic channels are disclosed herein. As disclosed, the cleaved amino acid residues may be immobilized in an array format and identified by detectable labels, such as antibodies, which specifically bind given amino acid residues. Alternatively, the antibodies are immobilized in an array format and the cleaved amino acids are labeled identified by being bound by the antibodies in the array.

  10. Immunoreagents and competitive assays to fludioxonil

    OpenAIRE

    Abad Fuentes, Antonio; Agulló, Consuelo; Esteve Turrillas, Francesc Albert; Abad Somovilla, Antonio; Mercader Badia, Josep Vicent

    2014-01-01

    Fludioxonil is a new-generation fungicide widely used for postharvest fruit protection. The aim of this study was to produce hitherto unreported immunoreagents for Fludioxonil analysis by immunoassay. Derivatives of this agrochemical were synthesized with different linker tethering sites. Those functionalized haptens were activated, and the purified active esters were efficiently conjugated to different carrier proteins for immunogen and assay antigen preparation. Antibodies to Fludioxonil we...

  11. Cell based assay for hypoglycemic drugs screening

    Institute of Scientific and Technical Information of China (English)

    LiZHANG; Juan-juanHU; Guan-huaDU

    2004-01-01

    OBJECTIVE: To establish a cell based assay for hypoglyc emicdrugs. METHODS: The five cell lines, BALB/c3T3, HepG2, NIH3T3, Be17402, and L929 were incubated with insulin (0-125n mol/L) for 48 h. Their sensitivities to insulin were studied by detecting glucose consumption. The dose-response and time-response relationship between the sensitive cell line (BALB/c 3T3)

  12. Polymerase chain reaction assay for avian polyomavirus.

    Science.gov (United States)

    Phalen, D N; Wilson, V G; Graham, D L

    1991-05-01

    A polymerase chain reaction assay was developed for detection of budgerigar fledgling disease virus (BFDV). The assay used a single set of primers complementary to sequences located in the putative coding region for the BFDV VP1 gene. The observed amplification product had the expected size of 550 bp and was confirmed to derive from BFDV DNA by its restriction digestion pattern. This assay was specific for BFDV and highly sensitive, being able to detect as few as 20 copies of the virus. By using the polymerase chain reaction, BFDV was detected in adult, nestling, and embryo budgerigar (Melopsitticus undulatus) tissue DNAs and in sera from adult and nestling budgerigars. These results suggest the possibility of persistent infections in adult birds and lend further support to previously described evidence of possible in ovo transmission. BFDV was also detected in chicken embryo fibroblast cell cultures and chicken eggs inoculated with the virus. A 550-bp product with identical restriction enzyme sites was amplified from a suspected polyomavirus isolated from a peach-faced lovebird (Agapornis pesonata) and from tissue DNA from a Hahn's macaw (Ara nobilis) and a sun conure (Aratinga solstitialis) with histological lesions suggestive of polyomavirus infection. These fragments also hybridized with a BFDV-derived probe, proving that they were derived from a polyomavirus very similar, if not identical, to BFDV.

  13. Diagnostic Certified Assay: Neuromuscular and Cardiac Assessments

    Directory of Open Access Journals (Sweden)

    Rea Valaperta

    2013-01-01

    Full Text Available The expansion of the specific trinucleotide sequence, [CTG], is the molecular pathological mechanism responsible for the clinical manifestations of DM1. Many studies have described different molecular genetic techniques to detect DM1, but as yet there is no data on the analytical performances of techniques used so far in this disease. We therefore developed and validated a molecular method, “Myotonic Dystrophy SB kit,” to better characterize our DM1 population. 113 patients were examined: 20 DM1-positive, 11 DM1/DM2-negative, and13 DM1-negative/DM2-positive, who had a previous molecular diagnosis, while 69 were new cases. This assay correctly identified 113/113 patients, and all were confirmed by different homemade assays. Comparative analysis revealed that the sensitivity and the specificity of the new kit were very high (>99%. Same results were obtained using several extraction procedures and different concentrations of DNA. The distribution of pathologic alleles showed a prevalence of the “classical” form, while of the 96 nonexpanded alleles 19 different allelic types were observed. Cardiac and neuromuscular parameters were used to clinically characterize our patients and support the new genetic analysis. Our findings suggest that this assay appears to be a very robust and reliable molecular test, showing high reproducibility and giving an unambiguous interpretation of results.

  14. Assessment of plaque assay methods for alphaviruses.

    Science.gov (United States)

    Juarez, Diana; Long, Kanya C; Aguilar, Patricia; Kochel, Tadeusz J; Halsey, Eric S

    2013-01-01

    Viruses from the Alphavirus genus are responsible for numerous arboviral diseases impacting human health throughout the world. Confirmation of acute alphavirus infection is based on viral isolation, identification of viral RNA, or a fourfold or greater increase in antibody titers between acute and convalescent samples. In convalescence, the specificity of antibodies to an alphavirus may be confirmed by plaque reduction neutralization test. To identify the best method for alphavirus and neutralizing antibody recognition, the standard solid method using a cell monolayer overlay with 0.4% agarose and the semisolid method using a cell suspension overlay with 0.6% carboxymethyl cellulose (CMC) overlay were evaluated. Mayaro virus, Una virus, Venezuelan equine encephalitis virus (VEEV), and Western equine encephalitis virus (WEEV) were selected to be tested by both methods. The results indicate that the solid method showed consistently greater sensitivity than the semisolid method. Also, a "semisolid-variant method" using a 0.6% CMC overlay on a cell monolayer was assayed for virus titration. This method provided the same sensitivity as the solid method for VEEV and also had greater sensitivity for WEEV titration. Modifications in plaque assay conditions affect significantly results and therefore evaluation of the performance of each new assay is needed.

  15. Hyperpolarized NMR Probes for Biological Assays

    Directory of Open Access Journals (Sweden)

    Sebastian Meier

    2014-01-01

    Full Text Available During the last decade, the development of nuclear spin polarization enhanced (hyperpolarized molecular probes has opened up new opportunities for studying the inner workings of living cells in real time. The hyperpolarized probes are produced ex situ, introduced into biological systems and detected with high sensitivity and contrast against background signals using high resolution NMR spectroscopy. A variety of natural, derivatized and designed hyperpolarized probes has emerged for diverse biological studies including assays of intracellular reaction progression, pathway kinetics, probe uptake and export, pH, redox state, reactive oxygen species, ion concentrations, drug efficacy or oncogenic signaling. These probes are readily used directly under natural conditions in biofluids and are often directly developed and optimized for cellular assays, thus leaving little doubt about their specificity and utility under biologically relevant conditions. Hyperpolarized molecular probes for biological NMR spectroscopy enable the unbiased detection of complex processes by virtue of the high spectral resolution, structural specificity and quantifiability of NMR signals. Here, we provide a survey of strategies used for the selection, design and use of hyperpolarized NMR probes in biological assays, and describe current limitations and developments.

  16. A single step protein assay that is both detergent and reducer compatible: The cydex blue assay.

    Science.gov (United States)

    Rabilloud, Thierry

    2016-10-01

    Determination of protein concentration is often an absolute prerequisite in preparing samples for biochemical and proteomic analyses. However, current protein assay methods are not compatible with both reducers and detergents, which are however present simultaneously in most denaturing extraction buffers used in proteomics and electrophoresis, and in particular in SDS electrophoresis. It was found that inclusion of cyclodextrins in a Coomassie blue-based assay made it compatible with detergents, as cyclodextrins complex detergents in a 1:1 molecular ratio. As this type of assay is intrinsically resistant to reducers, a single-step assay that is both detergent and reducer compatible was developed. Depending on the type and concentration of detergents present in the sample buffer, either beta-cyclodextrin or alpha-cyclodextrin can be used, the former being able to complex a wider range of detergents and the latter being able to complex higher amounts of detergents due to its greater solubility in water. Cyclodextrins are used at final concentrations of 2-10 mg/mL in the assay mix. This typically allows to measure samples containing as little as 0.1 mg/mL protein, in the presence of up to 2% detergent and reducers such as 5% mercaptoethanol or 50 mM DTT in a single step with a simple spectrophotometric assay.

  17. A novel assay detecting recall response to Mycobacterium tuberculosis: Comparison with existing assays.

    Science.gov (United States)

    Hsu, Denise C; Zaunders, John J; Plit, Marshall; Leeman, Craig; Ip, Susanna; Iampornsin, Thatri; Pett, Sarah L; Bailey, Michelle; Amin, Janaki; Ubolyam, Sasiwimol; Avihingsanon, Anchalee; Ananworanich, Jintanat; Ruxrungtham, Kiat; Cooper, David A; Kelleher, Anthony D

    2012-07-01

    A strategy to reduce the burden of active TB is isoniazid preventive therapy for latent TB infection (LTBI). However, current assays used to diagnose LTBI all have limitations. In these proof of concept studies, we compared the agreement of a novel flow cytometry assay detecting CD25/CD134 co-expression with QuantiFERON-TB Gold In-Tube (QFN-GIT) and Tuberculin skin test (TST) in the detection of recall immune response to TB. The CD25/CD134 assay, QFN-GIT and TST were performed on 74 participants referred for TB screening in Sydney and on 50 participants with advanced HIV infection (CD4 ≤ 350 × 10(6) cells/L) in Bangkok. The agreement between CD25/CD134 assay and QFN-GIT was 93.2% (Kappa 0.631 95% CI 0.336-0.926) in Sydney and 90% (Kappa 0.747 95% CI 0.541-0.954) in Bangkok. Discordant results occurred around the cut off of both tests. The agreement between CD25/CD134 assay and TST was 73.6% (Kappa 0.206 95% CI 0.004-0.409) in Sydney and 84% (Kappa 0.551 95% CI 0.296-0.806) in Bangkok. The CD25/CD134 assay showed good agreement with QFN-GIT in detecting recall response to TB both in well and less resourced setting as well as in persons with advanced HIV infection. Further study into the performance of this assay is thus warranted.

  18. Bacterial assay of contact lens wearers.

    Science.gov (United States)

    Hart, D E; Hosmer, M; Georgescu, M; Farris, R L

    1996-03-01

    The goal of the project was to determine the quantity of bacteria on the contact lens and adjacent areas of the eye. This paper is a quantitative study of the contact lens and ocular aerobic microbiota in a mixed group of daily and extended wear disposable contact lens users. The contact lens, the lower fornix, tears collecting at the lower fornix, and edge of the lower lid at the Meibomian gland margin were assayed for the quantity of bacterial colony forming units (CFU). Eighteen patients wearing 49 disposable high water content hydrogel contact lenses were assayed and the mean lens age was 8.8 +/- 4.6 days. Three patients wore their lenses on a daily wear basis and 15 on an extended wear schedule. Tear samples were obtained with sterile microbial loops and the lens was macerated into small particles with a tissue grinder. The samples were poured onto the surface of chocolate agar plates and incubated at 35 degrees C for 48 h in 5% Co2. The lid margin revealed the greatest bacterial presence (mean = 9.7 CFU; median = 2 CFU; mode = 0 CFU). The lens showed the next greatest presence of CFU (mean = 4.5 CFU; median = 1 CFU; mode = 0). The fornix and tears revealed the least bacterial presence (fornix: mean = 2.6 CFU; median = 0 CFU; mode = 0 CFU). The bacteria were coagulase-negative staphylococci. The bacterial assay of disposable lens wearing contact lens subjects indicates that the lid margins are the greatest source of bacteria with the tears being the lowest. These studies support the concept that in the eye, the lens typically does not possess a large number of bacteria under normal conditions.

  19. Nondestructive boxed transuranic (TRU) waste assay systems

    Science.gov (United States)

    Caldwell, John T.; Jones, Stephanie A.; Lucero, Randy F.

    1999-01-01

    A brief history of boxed waste assay systems (primarily those developed at Los Alamos National Laboratory) is presented. The characteristics and design process involved with current generation systems--as practiced by BII--are also discussed in some detail. Finally, a specific boxed waste assay system and acceptance test results are presented. This system was developed by BII and installed at the Waste Receiving and Packaging (WRAP) facility in Hanford, Washington in early 1997. The WRAP system combines imaging passive/active neutron (IPAN) techniques with gamma- ray energy analysis (GEA) to assay crates up to 2.5 m X 2.5 m X 6.5 m in size. (Systems that incorporate both these methodologies are usually denoted IPAN/GEA types.) Two separate gamma-ray measurements are accomplished utilizing 16 arrayed NaI detectors and a moveable HPGe detector, while 3He detectors acquire both active and passive neutron data. These neutron measurements use BII's proprietary imaging methodology. Acceptance testing of the system was conducted at Hanford in January 1998. The system's operating performance was evaluated based on accuracy and sensitivity requirements for three different matrix types. Test results indicate an average 13% active mode accuracy for 10 nCi/g loadings of Pu waste and 5% passive mode accuracy for 10 g loadings of Pu waste. Sensitivity testing demonstrated an active mode lower limit of detection of less than 5 nCi/g of 239Pu for the medium matrix and less than 20 pCi/g of fission and activation products at 3(sigma) above background.

  20. Antibacterial effect of protamine assayed by impedimetry

    DEFF Research Database (Denmark)

    Johansen, Charlotte; Gill, T.; Gram, Lone

    1995-01-01

    Impedimetric measurements were used to assay the antibacterial effect of protamine. A good linear correlation between the impedance detection time and the initial cell counts was obtained (r = 0 . 99, n = 2). As basic peptides may cause clumping of cells, this correlation curve was used when esti...... A suspended in buffer was not lethal as was the effect on growing cells; however, protamine (50-500 mu g ml(-1)) killed the Gram-negative fish spoilage bacteria Shewanella putrefaciens when the live cells were suspended in buffer....

  1. Test procedure for boxed waste assay system

    Energy Technology Data Exchange (ETDEWEB)

    Wachter, J. [Los Alamos National Lab., NM (United States)

    1994-12-07

    This document, prepared by Los Alamos National Laboratory`s NMT-4 group, details the test methodology and requirements for Acceptance/Qualification testing of a Boxed Waste Assay System (BWAS) designed and constructed by Pajarito Scientific Corporation. Testing of the BWAS at the Plutonium Facility (TA55) at Los Alamos National Laboratory will be performed to ascertain system adherence to procurement specification requirements. The test program shall include demonstration of conveyor handling capabilities, gamma ray energy analysis, and imaging passive/active neutron accuracy and sensitivity. Integral to these functions is the system`s embedded operating and data reduction software.

  2. Purification and kinase assay of PKN.

    Science.gov (United States)

    Mukai, Hideyuki; Ono, Yoshitaka

    2006-01-01

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

  3. Bicarbonate alters cellular responses in respiration assays.

    Science.gov (United States)

    Krycer, James R; Fisher-Wellman, Kelsey H; Fazakerley, Daniel J; Muoio, Deborah M; James, David E

    2017-08-05

    Metabolic assay buffers often omit bicarbonate, which is susceptible to alkalinisation in an open environment. Here, we assessed the effect of including bicarbonate in respirometry experiments. By supplementing HEPES-buffered media with low concentrations of bicarbonate, we found increased respiration in adipocytes and hepatocytes, but not myotubes. This was observed across multiple respirometry platforms and was independent of effects on enhanced insulin sensitivity, pH drift, or mitochondrial function. Permeabilised cell experiments suggest that bicarbonate increases substrate availability, likely by acting as a cofactor for carboxylase enzymes. This emphasises the importance of buffer choice in experimental biology. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Mass-based readout for agglutination assays

    Science.gov (United States)

    Chunara, Rumi; Godin, Michel; Knudsen, Scott M.; Manalis, Scott R.

    2007-11-01

    We present a mass-based readout for agglutination assays. The suspended microchannel resonator (SMR) is used to classify monomers and dimers that are formed during early stage aggregation, and to relate the total count to the analyte concentration. Using a model system of streptavidin functionalized microspheres and biotinylated antibody as the analyte, we obtain a dose-response curve over a concentration range of 0.63-630nM and show that the results are comparable to what has been previously achieved by image analysis and conventional flow cytometry.

  5. [Human angiogenin: expression, purification, biological assay].

    Science.gov (United States)

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

    2001-01-01

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

  6. Standardization of cytokine flow cytometry assays

    Directory of Open Access Journals (Sweden)

    Cox Josephine

    2005-06-01

    Full Text Available Abstract Background Cytokine flow cytometry (CFC or intracellular cytokine staining (ICS can quantitate antigen-specific T cell responses in settings such as experimental vaccination. Standardization of ICS among laboratories performing vaccine studies would provide a common platform by which to compare the immunogenicity of different vaccine candidates across multiple international organizations conducting clinical trials. As such, a study was carried out among several laboratories involved in HIV clinical trials, to define the inter-lab precision of ICS using various sample types, and using a common protocol for each experiment (see additional files online. Results Three sample types (activated, fixed, and frozen whole blood; fresh whole blood; and cryopreserved PBMC were shipped to various sites, where ICS assays using cytomegalovirus (CMV pp65 peptide mix or control antigens were performed in parallel in 96-well plates. For one experiment, antigens and antibody cocktails were lyophilised into 96-well plates to simplify and standardize the assay setup. Results (CD4+cytokine+ cells and CD8+cytokine+ cells were determined by each site. Raw data were also sent to a central site for batch analysis with a dynamic gating template. Mean inter-laboratory coefficient of variation (C.V. ranged from 17–44% depending upon the sample type and analysis method. Cryopreserved peripheral blood mononuclear cells (PBMC yielded lower inter-lab C.V.'s than whole blood. Centralized analysis (using a dynamic gating template reduced the inter-lab C.V. by 5–20%, depending upon the experiment. The inter-lab C.V. was lowest (18–24% for samples with a mean of >0.5% IFNγ + T cells, and highest (57–82% for samples with a mean of Conclusion ICS assays can be performed by multiple laboratories using a common protocol with good inter-laboratory precision, which improves as the frequency of responding cells increases. Cryopreserved PBMC may yield slightly more

  7. Premier`s Task Force on NAFTA wind turbine manufacturing facility and windpower plants : final report

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1998-03-01

    The proposal for a 700 MW wind farm and associated manufacturing facility at Pincher Creek, Alberta was discussed. York WindPower and ENERCON submitted the joint proposal in the spring of 1996 and requested a financial arrangement to guarantee a sale price of 5.4 cents per kWh (escalated) over 25 years. This was later revised in February, 1997 to 350 MW, 4.9 cents per kWh (escalated) over 15 years. A Task Force was established to assess this proposal and any other prospects for development of renewables in general. The two inseparable elements of the proposed project would be a manufacturing facility which would produce approximately 400 wind turbines a year, and a 700 MW wind farm, phased in over 10 years. The size of the wind farm would be based on a calculation of the minimal annual production required for the manufacturing facility to be viable. Pincher Creek residents are supportive of renewable energy and have been promoting wind energy long before the York/ENERCON proposal. They view this project as a support for regional economic development. The Task Force was pleased that York/ENERCON is pursuing the Alberta Advantage and is considering setting up a manufacturing facility. The Task Force did not seek to make a finding with respect to the business viability of the project, instead, the Task Force concluded that the commercial test for the project should be provided by competition in the marketplace. Nevertheless, the Task Force is supportive of the project, provided it proceeds on a voluntary, market-driven basis, and there are no conflicts with the existing government policy framework. 4 tabs., 2 appendices.

  8. Assaying environmental nickel toxicity using model nematodes.

    Science.gov (United States)

    Rudel, David; Douglas, Chandler D; Huffnagle, Ian M; Besser, John M; Ingersoll, Christopher G

    2013-01-01

    Although nickel exposure results in allergic reactions, respiratory conditions, and cancer in humans and rodents, the ramifications of excess nickel in the environment for animal and human health remain largely undescribed. Nickel and other cationic metals travel through waterways and bind to soils and sediments. To evaluate the potential toxic effects of nickel at environmental contaminant levels (8.9-7,600 µg Ni/g dry weight of sediment and 50-800 µg NiCl2/L of water), we conducted assays using two cosmopolitan nematodes, Caenorhabditis elegans and Pristionchus pacificus. We assayed the effects of both sediment-bound and aqueous nickel upon animal growth, developmental survival, lifespan, and fecundity. Uncontaminated sediments were collected from sites in the Midwestern United States and spiked with a range of nickel concentrations. We found that nickel-spiked sediment substantially impairs both survival from larval to adult stages and adult longevity in a concentration-dependent manner. Further, while aqueous nickel showed no adverse effects on either survivorship or longevity, we observed a significant decrease in fecundity, indicating that aqueous nickel could have a negative impact on nematode physiology. Intriguingly, C. elegans and P. pacificus exhibit similar, but not identical, responses to nickel exposure. Moreover, P. pacificus could be tested successfully in sediments inhospitable to C. elegans. Our results add to a growing body of literature documenting the impact of nickel on animal physiology, and suggest that environmental toxicological studies could gain an advantage by widening their repertoire of nematode species.

  9. Assaying environmental nickel toxicity using model nematodes.

    Directory of Open Access Journals (Sweden)

    David Rudel

    Full Text Available Although nickel exposure results in allergic reactions, respiratory conditions, and cancer in humans and rodents, the ramifications of excess nickel in the environment for animal and human health remain largely undescribed. Nickel and other cationic metals travel through waterways and bind to soils and sediments. To evaluate the potential toxic effects of nickel at environmental contaminant levels (8.9-7,600 µg Ni/g dry weight of sediment and 50-800 µg NiCl2/L of water, we conducted assays using two cosmopolitan nematodes, Caenorhabditis elegans and Pristionchus pacificus. We assayed the effects of both sediment-bound and aqueous nickel upon animal growth, developmental survival, lifespan, and fecundity. Uncontaminated sediments were collected from sites in the Midwestern United States and spiked with a range of nickel concentrations. We found that nickel-spiked sediment substantially impairs both survival from larval to adult stages and adult longevity in a concentration-dependent manner. Further, while aqueous nickel showed no adverse effects on either survivorship or longevity, we observed a significant decrease in fecundity, indicating that aqueous nickel could have a negative impact on nematode physiology. Intriguingly, C. elegans and P. pacificus exhibit similar, but not identical, responses to nickel exposure. Moreover, P. pacificus could be tested successfully in sediments inhospitable to C. elegans. Our results add to a growing body of literature documenting the impact of nickel on animal physiology, and suggest that environmental toxicological studies could gain an advantage by widening their repertoire of nematode species.

  10. Response definition criteria for ELISPOT assays revisited.

    Science.gov (United States)

    Moodie, Z; Price, L; Gouttefangeas, C; Mander, A; Janetzki, S; Löwer, M; Welters, M J P; Ottensmeier, C; van der Burg, S H; Britten, Cedrik M

    2010-10-01

    No consensus has been reached on how to determine if an immune response has been detected based on raw data from an ELISPOT assay. The goal of this paper is to enable investigators to understand and readily implement currently available methods for response determination. We describe empirical and statistical approaches, identifying the strengths and limitations of each approach to allow readers to rationally select and apply a scientifically sound method appropriate to their specific laboratory setting. Five representative approaches were applied to data sets from the CIMT Immunoguiding Program and the response detection and false positive rates were compared. Simulation studies were also performed to compare empirical and statistical approaches. Based on these, we recommend the use of a non-parametric statistical test. Further, we recommend that six medium control wells or four wells each for both medium control and experimental conditions be performed to increase the sensitivity in detecting a response, that replicates with large variation in spot counts be filtered out, and that positive responses arising from experimental spot counts below the estimated limit of detection be interpreted with caution. Moreover, a web-based user interface was developed to allow easy access to the recommended statistical methods. This interface allows the user to upload data from an ELISPOT assay and obtain an output file of the binary responses.

  11. Standardization of anti-DNA antibody assays.

    Science.gov (United States)

    Pisetsky, David S

    2013-07-01

    Antibodies to DNA (anti-DNA) are the serological hallmark of systemic lupus erythematosus and represent important biomarkers for clinical and research purposes. These antibodies are part of a family of antibodies to nucleosomes and bind to conserved sites widely present on DNA. While the value of anti-DNA as a biomarker is well established, the assay for these antibodies has involved a variety of DNA sources and systems to detect DNA-anti-DNA interactions. The influence of these variations on antibody detection has complicated assay standardization. As an antigen, DNA has unique features since it is a highly charged polymer that has structural heterogeneity. This heterogeneity can affect antigenicity which can vary on the basis of DNA origin, size, conformation and mobility. In addition, as a polymer, DNA can promote patterns of antibody binding based on monogamous or bivalent interaction which require an extended polynucleotide structure. Understanding the nature of DNA as an antigen can facilitate interpretation of serological tests and underpin efforts at better standardization.

  12. Assaying environmental nickel toxicity using model nematodes

    Science.gov (United States)

    Rudel, David; Douglas, Chandler; Huffnagle, Ian; Besser, John M.; Ingersoll, Christopher G.

    2013-01-01

    Although nickel exposure results in allergic reactions, respiratory conditions, and cancer in humans and rodents, the ramifications of excess nickel in the environment for animal and human health remain largely undescribed. Nickel and other cationic metals travel through waterways and bind to soils and sediments. To evaluate the potential toxic effects of nickel at environmental contaminant levels (8.9-7,600 µg Ni/g dry weight of sediment and 50-800 µg NiCl2/L of water), we conducted assays using two cosmopolitan nematodes, Caenorhabditis elegans and Pristionchus pacificus. We assayed the effects of both sediment-bound and aqueous nickel upon animal growth, developmental survival, lifespan, and fecundity. Uncontaminated sediments were collected from sites in the Midwestern United States and spiked with a range of nickel concentrations. We found that nickel-spiked sediment substantially impairs both survival from larval to adult stages and adult longevity in a concentration-dependent manner. Further, while aqueous nickel showed no adverse effects on either survivorship or longevity, we observed a significant decrease in fecundity, indicating that aqueous nickel could have a negative impact on nematode physiology. Intriguingly, C. elegansand P. pacificus exhibit similar, but not identical, responses to nickel exposure. Moreover, P. pacificus could be tested successfully in sediments inhospitable to C. elegans. Our results add to a growing body of literature documenting the impact of nickel on animal physiology, and suggest that environmental toxicological studies could gain an advantage by widening their repertoire of nematode species.

  13. Cryptosporidium cell culture infectivity assay design.

    Science.gov (United States)

    King, B J; Keegan, A R; Robinson, B S; Monis, P T

    2011-05-01

    Members of the genus Cryptosporidium, which cause the gastrointestinal disease cryptosporidiosis, still represent a significant cause of water-borne disease worldwide. While intensive efforts have been invested in the development of techniques for parasite culture, in vitro growth has been hampered by a number of factors including low levels of infectivity as well as delayed life-cycle development and poor synchronicity. In this study we examined factors affecting the timing of contact between excysted sporozoites and target host cells and the subsequent impact of this upon the establishment of infection. We demonstrate that excystation rate impacts upon establishment of infection and that in our standard assay format the majority of sporozoites are not close enough to the cell monolayer when they are released from the oocyst to successfully establish infection. However, this can be easily overcome by centrifugation of oocysts onto the cell monolayer, resulting in approximately 4-fold increases in sporozoite attachment and subsequent infection. We further demonstrate that excystation procedures can be tailored to control excystation rate to match the assay end purpose and that excystation rate can influence data interpretation. Finally, the addition of both a centrifugation and washing step post-sporozoite attachment may be appropriate when considering the design of in vitro culture experiments for developmental analysis and stage-specific gene expression as this appears to increase the synchronicity of early developmental stages.

  14. ACUTE ENTERIC INFECTIONS POLYMERASE CHAIN REACTION ASSAY IN PEDIATRIC PRACTICE: OPPORTUNITIES AND CHALLENGES

    Directory of Open Access Journals (Sweden)

    E. D. Sokolova

    2016-01-01

    Full Text Available The aim of the study is estimate the opportunities of local multi-prime PCR reagents kits in children enteric infections etiological diagnostics amongst the patients with diarrhoea vs traditional bacteriological methods. We used 4 kits of reagents that provide multiple pathogens simultaneous indication in one sample: 1 Rotavirus, Norovirus, Astrovirus; 2 Shigella spp./EIEC, Salmonella spp., Campylobacter spp.; 3 Yersinia enterocolitica and Yersinia pseudotuberculosis; 4 E. coli: EIEC (enteroinvasive, EPEC (enteropathogenic, ETEC (enterotoxigenic, EHEC (enterohaemorrhagic, EAgEC (enteroaggregative. It has been shown that the viral intestinal infections is increased by 14%, bacterial — in 2,5 times. PCR diagnostics identified in 62% of patients the viral gastroenteritis: Rotavirus (52%, Norovirus (9%, Astrovirus (1%. Detected bacterial pathogens PCR markers number proved up to 2.5 times high than according to bacteriological examination. The spectrum of bacterial agents increased due to E. coli and Y. enterocolitica. PCR diagnostics increased detection of Campylobacter up to 2 times. Detected E. coli DNA prevalence: EPEC — 66%, EAgEC, ETEC and EHEC were 31%, 9% and 4%, respectively. DNA Campylobacter spp. and E. coli constituted 2/3 of all findings: Campylobacter spp. (41%, E. coli (24%, Salmonella spp. (19%, Yersinia spp. (11%, Shigella spp./EIEC (5%. The positive results of bacteriological and serological methods duplicate the positive results of PCR diagnostics. In general, the positive results of PCR diagnosis of bacterial pathogens were detected in 46.35% of the examined patients. In 48.4% of patients identified PCR markers viral — bacterial infection, in 5.25% — of bacterial associations, in 11% of them were found the DNA 2–3 bacterial pathogens. The study was shown in children in St. Petersburg in 2012–2014 dominated rotavirus infection, campylobacteriosis and escherichiosis. The prevalence of viral-bacterial confections is more

  15. Fundamental equations for two-phase flow. Part 1: general conservation equations. Part 2: complement and remarks; Equations fondamentales des ecoulements diphasiques. Premiere partie: equations generales de conservation. Deuxieme partie: complements et remarques

    Energy Technology Data Exchange (ETDEWEB)

    Delhaye, J.M. [Commissariat a l' Energie Atomique, 38 - Grenoble (France). Centre d' Etudes Nucleaires

    1968-12-01

    equations concerning different methods of presenting two-phase flows with those proposed in the literature. As a result a detailed study has been made of the work of C.G. TELETOV, S.S. KUTATELADZE, M.A. STYRIKOVICH and N. ZUBER; a summary of this work presented. (author) [French] Ce rapport traite des equations generales de conservation de la masse, de la quantite de mouvement et de l'energie pour un ecoulement diphasique. Ces equations sont presentees sous plusieurs formes a partir des equations integrales qui sont posees a priori. 1. Equations aux variables locales instantanees et conditions d'interface; 2. Equations aux variables instantanees moyennees dans une section et applications pratiques: ces equations renferment une donnee experimentale interessante qui est le rapport de la section de passage d'une phase sur la section totale d'une conduite. 3. Equations aux moyennes statistiques locales et equations moyennees dans le temps: une tentative plus poussee pour relier experience et theorie consiste a prendre les moyennes statistiques des equations locales. On obtient alors des equations ou interviennent des variables moyennees dans le temps par application d'une hypothese ergodique. 4. Combinaisons des moyennes statistiques et des moyennes dans une section: on considere dans cette etude des variables locales moyennees statistiquement, puis moyennees dans la section et egalement des variables moyennees dans la section puis moyennees statistiquement. 5. Equations generales relatives aux emulsions: dans ce cas, une phase se presente (localement sous un aspect tres divise. Cette particularite permet de definir une concentration volumique locale et d'etablir des equations aux applications multiples. On a complete et precise certains points de la premiere partie de ce rapport concernant les equations generales de conservation des ecoulements diphasiques. On a introduit dans les equations generales les termes correspondant a la tension

  16. A Spectrophotometric Assay Optimizing Conditions for Pepsin Activity.

    Science.gov (United States)

    Harding, Ethelynda E.; Kimsey, R. Scott

    1998-01-01

    Describes a laboratory protocol optimizing the conditions for the assay of pepsin activity using the Coomasie Blue dye binding assay of protein concentration. The dye bonds through strong, noncovalent interactions to basic and aromatic amino acid residues. (DDR)

  17. Assay Method for 235U in Low-Density Waste

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    <正>235U assay method will provide a semi-quantitative assay for any uranium lumps that might exist in low-density, low-Z material waste boxes within a short count time. These materials will consist of

  18. Is the Comet Assay a Sensitive Procedure for Detecting Genotoxicity?

    Directory of Open Access Journals (Sweden)

    Satomi Kawaguchi

    2010-01-01

    Full Text Available Although the Comet assay, a procedure for quantitating DNA damage in mammalian cells, is considered sensitive, it has never been ascertained that its sensitivity is higher than the sensitivity of other genotoxicity assays in mammalian cells. To determine whether the power of the Comet assay to detect a low level of genotoxic potential is superior to those of other genotoxicity assays in mammalian cells, we compared the results of Comet assay with those of micronucleus test (MN test. WTK1 human lymphoblastoid cells were exposed to methyl nitrosourea (MNU, ethyl nitrosourea (ENU, methyl methanesulfonate (MMS, ethyl methanesulfonate (EMS, bleomycin (BLM, or UVC. In Comet assay, cells were exposed to each mutagen with (Comet assay/araC and without (Comet assay DNA repair inhibitors (araC and hydroxyurea. Furthermore, acellular Comet assay (acellular assay was performed to determine how single-strand breaks (SSBs as the initial damage contributes to DNA migration and/or to micronucleus formation. The lowest genotoxic dose (LGD, which is defined as the lowest dose at which each mutagen causes a positive response on each genotoxicity assay, was used to compare the power of the Comet assay to detect a low level of genotoxic potential and that of MN test; that is, a low LGD indicates a high power. Results are summarized as follows: (1 for all mutagens studied, LGDs were MN test ≦ Comet assay; (2 except for BLM, LGDs were Comet assay/araC ≦ MN test; (3 except for UVC and MNU, LGDs were acellular assay ≦ Comet assay/araC ≦ MN test ≦ Comet assay. The following is suggested by the present findings: (1 LGD in the Comet assay is higher than that in MN test, which suggests that the power of the MN test to detect a low level of genotoxic potential is superior to that of the Comet assay; (2 for the studied mutagens, all assays were able to detect all mutagens correctly, which suggests that the sensitivity of the Comet assay and that of the MN test were

  19. Assay optimization: a statistical design of experiments approach.

    Science.gov (United States)

    Altekar, Maneesha; Homon, Carol A; Kashem, Mohammed A; Mason, Steven W; Nelson, Richard M; Patnaude, Lori A; Yingling, Jeffrey; Taylor, Paul B

    2007-03-01

    With the transition from manual to robotic HTS in the last several years, assay optimization has become a significant bottleneck. Recent advances in robotic liquid handling have made it feasible to reduce assay optimization timelines with the application of statistically designed experiments. When implemented, they can efficiently optimize assays by rapidly identifying significant factors, complex interactions, and nonlinear responses. This article focuses on the use of statistically designed experiments in assay optimization.

  20. Performance Characteristics of Xpert Flu/RSV XC Assay.

    Science.gov (United States)

    Popowitch, Elena B; Miller, Melissa B

    2015-08-01

    The Xpert Flu/RSV XC assay was compared to laboratory-developed tests (LDTs) (n = 207) and the Xpert Flu assay (n = 147) using archived nasopharyngeal swabs. The percentages of positive agreements with LDTs were 97.8% for influenza A, 97.2% for influenza B, and 89.3% for RSV. The sensitivity of influenza detection was improved with the Xpert Flu/RSV XC assay compared to the Xpert Flu assay.

  1. Comparison of two rapid assays for Clostridium difficile Common antigen and a C difficile toxin A/B assay with the cell culture neutralization assay.

    Science.gov (United States)

    Reller, Megan E; Alcabasa, Romina C; Lema, Clara A; Carroll, Karen C

    2010-01-01

    We compared 3 rapid assays for Clostridium difficile with a cell culture cytotoxicity neutralization assay (CCNA). Of 600 stool samples, 46 were positive for toxigenic C difficile. Both rapid common antigen assays were highly sensitive (91.3%-100%) and, therefore, were appropriate screening tests. The rapid toxin assay had poor sensitivity (61%) but excellent specificity (99.3%). Testing stools for glutamate dehydrogenase (step 1) and those positive with a rapid toxin assay (step 2) would correctly classify 81% of submitted specimens within 2 hours, including during periods of limited staffing (evenings, nights, and weekends). CCNA could then be used as a third step to test rapid toxin-negative samples, thereby providing a final result for the remaining 19% of samples by 48 to 72 hours. The use of rapid assays as outlined could enhance timely diagnosis of C difficile.

  2. Systems, devices, and methods for agglutination assays using sedimentation

    Science.gov (United States)

    Schaff, Ulrich Y.; Sommer, Gregory J.; Singh, Anup K.

    2016-01-26

    Embodiments of the present invention include methods for conducting agglutination assays using sedimentation. Aggregates may be exposed to sedimentation forces and travel through a density medium to a detection area. Microfluidic devices, such as microfluidic disks, are described for conducting the agglutination assays, as are systems for conducting the assays.

  3. 21 CFR 866.2350 - Microbiological assay culture medium.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Microbiological assay culture medium. 866.2350... Microbiological assay culture medium. (a) Identification. A microbiological assay culture medium is a device that... organism in the innoculated medium. Test results aid in the diagnosis of disease resulting from either...

  4. 21 CFR 864.7470 - Glycosylated hemoglobin assay.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Glycosylated hemoglobin assay. 864.7470 Section... Glycosylated hemoglobin assay. (a) Identification. A glycosylated hemoglobin assay is a device used to measure the glycosylated hemoglobins (A1a, A1b, and A1c) in a patient's blood by a column...

  5. 21 CFR 864.7400 - Hemoglobin A2 assay.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Hemoglobin A2 assay. 864.7400 Section 864.7400...) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7400 Hemoglobin A2 assay. (a) Identification. A hemoglobin A2 assay is a device used to determine the hemoglobin A2...

  6. 21 CFR 864.7500 - Whole blood hemoglobin assays.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Whole blood hemoglobin assays. 864.7500 Section... blood hemoglobin assays. (a) Identification. A whole blood hemoglobin assay is a device consisting or... hemoglobin content of whole blood for the detection of anemia. This generic device category does not...

  7. 21 CFR 864.7415 - Abnormal hemoglobin assay.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Abnormal hemoglobin assay. 864.7415 Section 864... hemoglobin assay. (a) Identification. An abnormal hemoglobin assay is a device consisting of the reagents... hemoglobin types. (b) Classification. Class II (performance standards)....

  8. 21 CFR 866.3402 - Plasmodium species antigen detection assays.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Plasmodium species antigen detection assays. 866... Plasmodium species antigen detection assays. (a) Identification. A Plasmodium species antigen detection assay... malaria caused by the four malaria species capable of infecting humans: Plasmodium falciparum, Plasmodium...

  9. 40 CFR 79.64 - In vivo micronucleus assay.

    Science.gov (United States)

    2010-07-01

    ... micronucleus assay. (a) Purpose. The micronucleus assay is an in vivo cytogenetic test which uses erythrocytes...) Test evaluation. (i) Positive results in the micronucleus test provide information on the ability of a..., Mammalian Bone Marrow Cytogenetics Tests: Micronucleus Assay. (2) Cihak, R. “Evaluation of Benzidine by...

  10. 21 CFR 866.3305 - Herpes simplex virus serological assays.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Herpes simplex virus serological assays. 866.3305... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3305 Herpes simplex virus serological assays. (a) Identification. Herpes simplex virus serological assays are devices...

  11. 21 CFR 864.7100 - Red blood cell enzyme assay.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Red blood cell enzyme assay. 864.7100 Section 864...) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7100 Red blood cell enzyme assay. (a) Identification. Red blood cell enzyme assay is a device used to measure the activity...

  12. Assays for investigating deSUMOylation enzymes.

    Science.gov (United States)

    Madu, Ikenna G; Chen, Yuan

    2012-07-01

    Post-translational modifications by the SUMO (Small Ubiquitin-like MOdifier) family of proteins are recently discovered essential regulatory mechanisms. All SUMO proteins are synthesized as larger precursors that are matured by SUMO-specific proteases, known as SENPs, which remove several C-terminal amino acids of SUMO to expose the Gly-Gly motif. SENPs also remove SUMO modifications from target proteins, making this modification highly dynamic. At least six deSUMOylation enzymes, all of which are encoded by essential genes, have been identified in mammals. SENP1 has been shown to play an important role in the development of prostate cancer and in angiogenesis. This unit describes and discusses methods for characterizing the deSUMOylation enzymes. These assays enable the identification of inhibitors of these enzymes and investigation of their mechanism of inhibition in order to develop research tools and future therapeutics.

  13. Nondestructive Assay Options for Spent Fuel Encapsulation

    Energy Technology Data Exchange (ETDEWEB)

    Tobin, Stephen J. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Jansson, Peter [Uppsala Univ. (Sweden)

    2014-10-02

    This report describes the role that nondestructive assay (NDA) techniques and systems of NDA techniques may have in the context of an encapsulation and deep geological repository. The potential NDA needs of an encapsulation and repository facility include safeguards, heat content, and criticality. Some discussion of the facility needs is given, with the majority of the report concentrating on the capability and characteristics of individual NDA instruments and techniques currently available or under development. Particular emphasis is given to how the NDA techniques can be used to determine the heat production of an assembly, as well as meet the dual safeguards needs of 1) determining the declared parameters of initial enrichment, burn-up, and cooling time and 2) detecting defects (total, partial, and bias). The report concludes with the recommendation of three integrated systems that might meet the combined NDA needs of the encapsulation/repository facility.

  14. Assay of Endocannabinoid Oxidation by Cyclooxygenase-2

    Science.gov (United States)

    Kudalkar, Shalley N.; Kingsley, Philip J.; Marnett, Lawrence J.

    2017-01-01

    Summary The endocannabinoids, 2-arachidonoylglycerol (2-AG) and arachidonylethanolamide (AEA) are endogenous ligands for the cannabinoid receptors (CB1 and CB2) and are implicated in a wide array of physiological processes. These neutral arachidonic acid (AA) derivatives have been identified as efficient substrates for the second isoform of the cyclooxygenase enzyme (COX-2). A diverse family of prostaglandin glycerol esters (PG-Gs) and prostaglandin ethanolamides (PG-EAs) is generated by the action of COX-2 (and downstream prostaglandin synthases) on 2-AG and AEA. As the biological importance of the endocannabinoid system becomes more apparent, there is a tremendous need for robust, sensitive and efficient analytical methodology for the endocannabinoids and their metabolites. In this chapter we describe methodology suitable for carrying out oxygenation of endocannabinoids by COX-2, and analysis of products of endocannabinoid oxygenation by COX-2 and of endocannabinoids themselves from in-vitro and cell assays. PMID:27245906

  15. Stable isotope dilution assays in mycotoxin analysis

    Energy Technology Data Exchange (ETDEWEB)

    Rychlik, Michael; Asam, Stefan [Universitaet Muenchen, Lehrstuhl fuer Lebensmittelchemie der Technischen, Garching (Germany)

    2008-01-15

    The principle and applications of stable isotope dilution assays (SIDAs) in mycotoxin analysis are critically reviewed. The general section includes historical aspects of SIDAs, the prerequisites and limitations of the use of stable isotopically labelled internal standards, and possible calibration procedures. In the application section actual SIDAs for the analysis of trichothecenes, zearalenone, fumonisins, patulin, and ochratoxin A are presented. The syntheses and availability of labelled mycotoxins for use as internal standards is reviewed and specific advances in food analysis and toxicology are demonstrated. The review indicates that LC-MS applications, in particular, require the use of stable isotopically labelled standards to compensate for losses during clean-up and for discrimination due to ion suppression. As the commercial availability of these compounds continues to increase, SIDAs can be expected to find expanding use in mycotoxin analysis. (orig.)

  16. Lab-on-a-Chip Multiplex Assays.

    Science.gov (United States)

    Peter, Harald; Wienke, Julia; Bier, Frank F

    2017-01-01

    Lab-on-a-chip multiplex assays allow a rapid identification of multiple parameters in an automated manner. Here we describe a lab-based preparation followed by a rapid and fully automated DNA microarray hybridization and readout in less than 10 min using the Fraunhofer in vitro diagnostics (ivD) platform to enable rapid identification of bacterial species and detection of antibiotic resistance. The use of DNA microarrays allows a fast adaptation of new biomarkers enabling the identification of different genes as well as single-nucleotide-polymorphisms (SNPs) within these genes. In this protocol we describe a DNA microarray developed for identification of Staphylococcus aureus and the mecA resistance gene.

  17. Development of a Radioactive Waste Assay System

    Energy Technology Data Exchange (ETDEWEB)

    Kang, Duck Won; Song, Myung Jae; Shin, Sang Woon; Sung, Kee Bang; Ko, Dae Hach [Korea Electric Power Research Institute, Taejon (Korea, Republic of); Kim, Kil Jeong; Park, Jong Mook; Jee, Kwang Yoong [Korea Atomic Energy Research Institute, Taejon (Korea, Republic of)

    1996-12-31

    Nuclear Act of Korea requires the manifest of low and intermediate level radioactive waste generated at nuclear power plants prior to disposal sites.Individual history records of the radioactive waste should be contained the information about the activity of nuclides in the drum, total activity, weight, the type of waste. A fully automated nuclide analysis assay system, non-destructive analysis and evaluation system of the radioactive waste, was developed through this research project. For the nuclides that could not be analysis directly by MCA, the activities of the representative {gamma}-emitters(Cs-137, Co-60) contained in the drum were measured by using that system. Then scaling factors were used to calculate the activities of {alpha}, {beta}-emitters. Furthermore, this system can automatically mark the analysis results onto the drum surface. An automated drum handling system developed through this research project can reduce the radiation exposure to workers. (author). 41 refs., figs.

  18. Stable isotope dilution assays in mycotoxin analysis.

    Science.gov (United States)

    Rychlik, Michael; Asam, Stefan

    2008-01-01

    The principle and applications of stable isotope dilution assays (SIDAs) in mycotoxin analysis are critically reviewed. The general section includes historical aspects of SIDAs, the prerequisites and limitations of the use of stable isotopically labelled internal standards, and possible calibration procedures. In the application section actual SIDAs for the analysis of trichothecenes, zearalenone, fumonisins, patulin, and ochratoxin A are presented. The syntheses and availability of labelled mycotoxins for use as internal standards is reviewed and specific advances in food analysis and toxicology are demonstrated. The review indicates that LC-MS applications, in particular, require the use of stable isotopically labelled standards to compensate for losses during clean-up and for discrimination due to ion suppression. As the commercial availability of these compounds continues to increase, SIDAs can be expected to find expanding use in mycotoxin analysis.

  19. Universal fieldable assay with unassisted visual detection

    Science.gov (United States)

    Chelyapov, Nicolas (Inventor)

    2012-01-01

    A universal detection system based on allosteric aptamers, signal amplification cascade, and eye-detectable phrase transition. A broadly applicable homogeneous detection system is provided. It utilizes components of the blood coagulation cascade in the presence of polystyrene microspheres (MS) as a signal amplifier. Russell's viper venom factor X activator (RVV-X) triggers the cascade, which results in an eye-visible phase transition--precipitation of MS bound to clotted fibrin. An allosteric RNA aptamer, RNA132, with affinity for RVV-X and human vascular endothelial growth factor (VEGF.sub.165) was created. RNA132 inhibits enzymatic activity of RVV-X. The effector molecule, VEGF.sub.165, reverses the inhibitory activity of RNA132 on RVV-X and restores its enzymatic activity, thus triggering the cascade and enabling the phase transition. Similar results were obtained for another allosteric aptamer modulated by a protein tyrosine phosphatase. The assay is instrumentation-free for both processing and readout.

  20. COMPARATIVE STUDY ON MILK CASEIN ASSAY METHODS

    Directory of Open Access Journals (Sweden)

    RODICA CĂPRIłĂ

    2013-12-01

    Full Text Available Casein, the main milk protein was determined by different assay methods: the gravimetric method, the method based on the neutralization of the NaOH excess used for the casein precipitate solving and the method based on the titration of the acetic acid used for the casein precipitation. The last method is the simplest one, with the fewer steps, and also with the lowest error degree. The results of the experiment revealed that the percentage of casein from the whole milk protein represents between 72.6–81.3% in experiment 1, between 73.6–81.3% in experiment 2 and between 74.3–81% in experiment 3.

  1. Enzymatic assay for methotrexate in erythrocytes

    DEFF Research Database (Denmark)

    Schrøder, H; Heinsvig, E M

    1985-01-01

    Methotrexate (MTX) accumulates in erythrocytes in MTX-treated patients. We present a modified enzymatic assay measuring MTX concentrations between 10 and 60 nmol/l in erythrocytes, adapted for a centrifugal analyser (Cobas Bio). About 40 patient's samples could be analysed within 1 h. The detection...... limit was 3 nmol/l. Within run and between-run precision was 7.4% and 13.5% for control 10 nmol/l and 1.2% and 3.2% for control 50 nmol/l. Recovery was 85-115% of MTX added to haemolysed erythrocytes. We found the method useful for pharmacokinetic studies of MTX in erythrocytes in MTX-treated patients...

  2. Advancement in biochemical assays in andrology

    Institute of Scientific and Technical Information of China (English)

    Wolf-BernhardSchill; RaftHenkel

    1999-01-01

    Determination of maikers of sperm function, accessory sex gland secretion and silent male genital tract inflammation is of considerable diagnostic value in the evaluation of male infertility. The introduction of biochemical tests into the analysis of male factor has the advantage that standardized assays with a coefficient of variafion characteristic of clinical chemistry are performed, in contrast to biological test systems with a large variability .Biochemical parameters may be used in clinical practice to evaluate the sperm fertitizing capacity (acrosin, aniline blue,ROS), to characterize male accessory sex gland secretinns (fructose, a-glucosidase, PSA), and to identify men with silent genital tract inflammation (elastase, C'3 complement component, coeruloplasmin, IgA, IgG, ROS). (As/an J Androl 1999 Jun; 1: 45-51)

  3. Medical Devices; Immunology and Microbiology Devices; Classification of the Assayed Quality Control Material for Clinical Microbiology Assays. Final order.

    Science.gov (United States)

    2017-07-27

    The Food and Drug Administration (FDA, Agency, or we) is classifying the assayed quality control material for clinical microbiology assays into class II (special controls). The special controls that will apply to the device are identified in this order and will be part of the codified language for the assayed quality control material for clinical microbiology assays' classification. The Agency is classifying the device into class II (special controls) to provide a reasonable assurance of safety and effectiveness of the device.

  4. Improved internal control for molecular diagnosis assays.

    Science.gov (United States)

    Vinayagamoorthy, T; Maryanski, Danielle; Vinayagamoorthy, Dilanthi; Hay, Katie S L; Yo, Jacob; Carter, Mark; Wiegel, Joseph

    2015-01-01

    The two principal determining steps in molecular diagnosis are the amplification and the identification steps. Accuracy of DNA amplification is primarily determined by the annealing sequence of the PCR primer to the analyte DNA. Accuracy for identification is determined either by the annealing region of a labelled probe for the real time PCR analysis, or the annealing of a sequencing primer for DNA sequencing analysis, that binds to the respective analyte (amplicon). Presently, housekeeping genes (Beta globin, GAPDH) are used in molecular diagnosis to verify that the PCR conditions are optimum, and are thus known as amplification controls [1-4]. Although these genes have been useful as amplification controls, they lack the true definition of an internal control because the primers and annealing conditions are not identical to the analyte being assayed. This may result in a false negative report [5]. The IC-Code platform technology described here provides a true internal control where the internal control and analyte share identical PCR primers annealing sequences for the amplification step and identical sequencing primer annealing sequence for the identification step. •The analyte and internal control have the same PCR and sequencing annealing sequences.•This method assures for little or no false negatives and false positives due to the method's design of using identical annealing conditions for the internal control and analyte, and by using DNA sequencing analysis for the identification step of the analyte, respectively.•This method also allows for a set lower limit of detection to be used by varying the amount of internal control used in the assay.

  5. Antioxidant assay using genetically engineered bioluminescent Escherichia coli

    Science.gov (United States)

    Bartolome, Amelita; Macalino, Bernadette; Pastoral, Ian Lemuel; Sevilla, Fortunato, III

    2006-02-01

    A new antioxidant activity assay based on the reactive oxygen species (ROS)-inducible bacterial strain (E. coli DPD2511) is described. The strain harbors the plasmid pKatG::luxCDABE and responds to hydrogen peroxide treatment by increasing light emission at 490 nm. Antioxidant capacity is evaluated through the ability of an agent to inhibit the hydrogen peroxide-induced bioluminescence of E. coli DPD2511. Applicability of the developed assay in detecting levels of antioxidants in various aqueous plant extracts is demonstrated. The assay was validated against 2,2-diphenylpicrylhydrazyl (DPPH) assay, a known antioxidant assay.

  6. Spectrophotometric Enzyme Assays for High-Throughput Screening

    Directory of Open Access Journals (Sweden)

    Jean-Louis Reymond

    2004-01-01

    Full Text Available This paper reviews high-throughput screening enzyme assays developed in our laboratory over the last ten years. These enzyme assays were initially developed for the purpose of discovering catalytic antibodies by screening cell culture supernatants, but have proved generally useful for testing enzyme activities. Examples include TLC-based screening using acridone-labeled substrates, fluorogenic assays based on the β-elimination of umbelliferone or nitrophenol, and indirect assays such as the back-titration method with adrenaline and the copper-calcein fluorescence assay for aminoacids.

  7. Performance of MycAssay Aspergillus DNA real-time PCR assay compared with the galactomannan detection assay for the diagnosis of invasive aspergillosis from serum samples.

    Science.gov (United States)

    Danylo, Alexis; Courtemanche, Chantal; Pelletier, René; Boudreault, Alexandre A

    2014-08-01

    Invasive aspergillosis (IA) is a major problem in the immunocompromised population, and its diagnosis is difficult due to the low sensitivity of available tests. Detection of Aspergillus nucleic acid by polymerase chain reaction (PCR) in serum samples is a promising diagnostic tool; however, use of multiple "in-house" methods precludes standardization. The first commercial PCR assay, MycAssay Aspergillus (Myconostica, Ltd), became available recently, and its performance in the diagnosis of IA was evaluated and compared with the galactomannan (GM) assay. Serum samples obtained from patients with hematological cancer were tested retrospectively with MycAssay Aspergillus PCR. Per-episode and per-test analyses were undertaken with 146 sera from 35 hematological patients. Sixteen patients had proven or probable IA and 19 had possible or no IA. In per-episode analysis, MycAssay Aspergillus had a sensitivity of 43.8% (95% confidence interval [CI], 19.8%-70.1%) and a specificity of 63.2% (95% CI, 38.4%-83.7%) for IA diagnosis. In per-test analyses, MycAssay Aspergillus had a lower specificity than the GM assay (83.3% vs. 93.1%, P = 0.04). The addition of PCR to routine clinical practice would have permitted the diagnosis of one additional probable IA in our cohort. Use of PCR instead of GM assay would have delayed the diagnosis in two cases. Aspergillus DNA detection by PCR with serum specimens using MycAssay showed a lower specificity than the GM assay and was associated with a low sensitivity for IA diagnosis. More studies are needed to determine the exact role of MycAssay in IA diagnosis in patients with hematological malignancy.

  8. Shiga toxins decrease enterohaemorrhagic Escherichia coli survival within Acanthamoeba castellanii.

    Science.gov (United States)

    Chekabab, Samuel M; Daigle, France; Charette, Steve J; Dozois, Charles M; Harel, Josée

    2013-07-01

    Enterohaemorrhagic Escherichia coli (EHEC) are zoonotic pathogens transmitted to humans through contaminated water or bovine products. One of the strategies used by pathogenic bacteria to survive in aquatic environments is using free-living amoebae as hosts. Acanthamoeba castellanii is an amoeba known to host several waterborne pathogens. This study investigates the survival of EHEC with A. castellanii, which could contribute to its spread and transmission to humans. We used a gentamicin protection assay as well as fluorescence and electron microscopy to monitor the intra-amoebae survival of EHEC O157:H7 over 24 h. The results showed that EHEC were able to survive within A. castellanii and that this survival was reduced by Shiga toxins (Stx) produced by EHEC. A toxic effect mediated by Stx was demonstrated by amoebae mortality and LDH release during co-culture of EHEC and amoeba. This work describes the ability of EHEC to survive within A. castellanii, and this host-pathogen interaction is partially controlled by the Stx. Thus, this ubiquitous amoeba could represent an environmental niche for EHEC survival and transmission.

  9. Technical note: comparison of the PrestoBlue and LDH release assays with the MTT assay for skin viability assessment.

    Science.gov (United States)

    Gaucher, Sonia; Jarraya, Mohamed

    2015-09-01

    MTT assay is the gold standard for assessing skin sample viability but it is time-consuming. Here we compared the MTT test with two other assays for the assessment of skin viability. The MTT, PrestoBlue (colorimetric method) and LDH release assays were applied to fresh and cryopreserved skin. Skin viability was considered proportional to the optical density values of the relevant analytes. PrestoBlue did not reliably distinguish between fresh and cryopreserved skin. The LDH release assay did not allow us to establish a viability index. We recommend the MTT assay for assessing skin viability.

  10. The low molecular weight DNA diffusion assay as an indicator of cytotoxicity for the in vitro comet assay.

    Science.gov (United States)

    Speit, Günter; Vesely, Alexandra; Schütz, Petra; Linsenmeyer, Regina; Bausinger, Julia

    2014-07-01

    The low molecular weight DNA diffusion assay (LMW assay) has been recommended as a measure for cytotoxicity for the in vivo comet assay. To better understand the relationship between effects in the LMW assay, DNA migration in the comet assay and effects in established cytotoxicity tests, we performed in vitro experiments with cultured human cell lines (TK6, A549) and comparatively investigated five test substances (methyl methanesulfonate, (±)-benzo[a]pyrene diol epoxide, sodium dodecyl sulphate, menthol and sodium arsenite). We measured DNA migration (tail intensity) in the comet assay and the frequency of 'hedgehogs' (cells with almost all DNA in the tail), DNA diffusion in the LMW assay, cell viability (trypan blue and fluorescein diacetate/ethidium bromide staining) and inhibition of proliferation (relative cell counts). Our in vitro experiments indicate that effects in the LMW assay occur independently from DNA effects in the comet assay and are not related to the occurrence of hedgehogs. Results from the LMW assay are in good agreement with results from viability assays and seem to allow discriminating genotoxic from non-genotoxic substances when appropriate preparation times are considered. Measurements of cytotoxicity by these methods only at an early preparation time after exposure to genotoxic substances may lead to erroneous results.

  11. Heated oligonucleotide ligation assay (HOLA): an affordable single nucleotide polymorphism assay.

    Science.gov (United States)

    Black, W C; Gorrochotegui-Escalante, N; Duteau, N M

    2006-03-01

    Most single nucleotide polymorphism (SNP) detection requires expensive equipment and reagents. The oligonucleotide ligation assay (OLA) is an inexpensive SNP assay that detects ligation between a biotinylated "allele-specific detector" and a 3' fluorescein-labeled "reporter" oligonucleotide. No ligation occurs unless the 3' detector nucleotide is complementary to the SNP nucleotide. The original OLA used chemical denaturation and neutralization. Heated OLA (HOLA) instead uses a thermal stable ligase and cycles of denaturing and hybridization for ligation and SNP detection. The cost per genotype is approximately US$1.25 with two-allele SNPs or approximately US$1.75 with three-allele SNPs. We illustrate the development of HOLA for SNP detection in the Early Trypsin and Abundant Trypsin loci in the mosquito Aedes aegypti (L.) and at the a-glycerophosphate dehydrogenase locus in the mosquito Anopheles gambiae s.s.

  12. Treatment of in vitro enterohemorrhagic Escherichia coli infection using phage and probiotics.

    Science.gov (United States)

    Dini, C; Bolla, P A; de Urraza, P J

    2016-07-01

    To assay the combination of phage and probiotics against EHEC in vitro on infected Hep-2 cells. Phage and probiotics treatments on EHEC O157:H7-infected Hep-2 cells were assayed individually or combined. The effect of freeze-drying on phage and probiotic antimicrobial activity was also studied. While treatment with phage alone increased cell detachment caused by EHEC infection, the treatments with MM alone or in combination with phage proved to effectively diminish cell damage caused by EHEC infection. Combined treatment showed a decrease in apoptotic cell count of 57·3% and a reduction in EHEC adhesion to cell monolayer of 1·2 log CFU. The simultaneous use of phage and probiotics showed no antagonistic effect, and freeze-drying did not affect their antipathogenic activity. The combination of phage and probiotics has great potential for reducing the number of pathogens adhered to epithelial cells during EHEC O157:H7 infection and attenuating the cytotoxic effect derived from it. Further in vivo assays are needed for assessing the actual effectiveness of the treatment. This study presents a freeze-dried formulation of phage and probiotics capable of controlling EHEC infections and reducing epithelial cell damage in vitro. © 2016 The Society for Applied Microbiology.

  13. Repeated dose liver micronucleus assay using adult mice with multiple genotoxicity assays concurrently performed as a combination test.

    Science.gov (United States)

    Hagio, Soichiro; Furukawa, Satoshi; Abe, Masayoshi; Kuroda, Yusuke; Hayashi, Seigo; Ogawa, Izumi

    2014-06-01

    Recently, the liver micronucleus (MN) assay using young adult rats with repeated administrations has been investigated by employing a new method without partial hepatectomy or in situcollagenase perfusion as the repeated dose liver MN (RDLMN) assay by Narumi et al. (2012). In our study, in order to investigate the possibility of the RDLMN assay using young adult mice instead of rats and the feasibility of employing some genotoxicity assays along with the RDLMN assay as a combination test, two genotoxic carcinogens (N,N-diethylnitrosoamine (DEN) and cisplatin (CIS)) and a nongenotoxic carcinogen (phenobarbital sodium (PHE)) were administered to mice for 15 or 29 days. Then, the liver MN assay, peripheral blood (PB) MN assay and comet assay using the liver and kidney were concurrently performed as a combination test. DEN showed positive responses to all endpoints except MN induction in PB after 15 days of repeat administration. A cross-linking agent, CIS, showed MN induction in liver after 29 days of repeat administration, and in PB after 15 and 29 days of repeat administration, although the comet assay yielded negative responses for both organs at both sampling times. PHE yielded negative responses for all endpoints. In conclusion, it is suggested that the RDLMN assay using mice is a feasible method to be integrated into the general repeated toxicity test along with the combination assays, i.e., comet assay or PB MN assay, which would help in risk assessment for carcinogenicity by comparing the results of combination assays with each other.

  14. Fluoride assay methodology for carbonated beverages.

    Science.gov (United States)

    Heilman, Judith R; Levy, Steven M; Wefel, James S; Patterson, Kristine Y; Cutrufelli, Rena; Pehrsson, Pamela R; Holden, Joanne M

    2006-01-01

    The purpose of this paper was to review different methodological techniques used for the assessment of fluoride in carbonated beverages, and compare results using a fluoride ion electrode direct read method with and without a prior decarbonation treatment. The carbonated beverages in this study were either purchased locally at grocery stores in Iowa City, Iowa, or purchased as part of a national representative sampling approach included in the National Fluoride Database and Intake Assessment Study (NFDIAS). The samples were compared with and without a decarbonating process. Soda pop and beer samples were analyzed by removing a 1-ml sample and adding a 1-ml buffer solution. The fluoride concentration of the sample and buffer combination was then determined using a fluoride ion specific electrode. There was no significant difference in the fluoride concentration of the samples with or without prior decarbonation. The mean absolute difference between the soda pop group with and without decarbonation was 0.01 ppm F, while results from the beer samples showed variation of 0.00 to 0.02 parts per million fluoride (ppm F). These differences were not statistically significant for the soda pop or beer groups (P=.50 and P=.74, respectively). Whether or not decarbonation was conducted prior to analysis, the fluoride assay results were the same. Therefore, decarbonation of soda pop and beer was deemed unnecessary prior to fluoride analysis.

  15. The synchronous active neutron detection assay system

    Energy Technology Data Exchange (ETDEWEB)

    Pickrell, M.M.; Kendall, P.K.

    1994-08-01

    We have begun to develop a novel technique for active neutron assay of fissile material in spent nuclear fuel. This approach will exploit a 14-MeV neutron generator developed by Schlumberger. The technique, termed synchronous active neutron detection (SAND), follows a method used routinely in other branches of physics to detect very small signals in presence of large backgrounds. Synchronous detection instruments are widely available commercially and are termed ``lock-in`` amplifiers. We have implemented a digital lock-in amplifier in conjunction with the Schlumberger neutron generator to explore the possibility of synchronous detection with active neutrons. The Schlumberger system can operate at up to a 50% duty factor, in effect, a square wave of neutron yield. Results are preliminary but promising. The system is capable of resolving the fissile material contained in a small fraction of the fuel rods in a cold fuel assembly; it also appears resilient to background neutron interference. The interrogating neutrons appear to be non-thermal and penetrating. Work remains to fully explore relevant physics and optimize instrument design.

  16. Prandiology of Drosophila and the CAFE assay

    Science.gov (United States)

    Ja, William W.; Carvalho, Gil B.; Mak, Elizabeth M.; de la Rosa, Noelle N.; Fang, Annie Y.; Liong, Jonathan C.; Brummel, Ted; Benzer, Seymour

    2007-01-01

    Studies of feeding behavior in genetically tractable invertebrate model systems have been limited by the lack of proper methodology. We introduce the Capillary Feeder (CAFE), a method allowing precise, real-time measurement of ingestion by individual or grouped fruit flies on the scale of minutes to days. Using this technique, we conducted the first quantitative analysis of prandial behavior in Drosophila melanogaster. Our results allow the dissection of feeding into discrete bouts of ingestion, defining two separate parameters, meal volume and frequency, that can be uncoupled and thus are likely to be independently regulated. In addition, our long-term measurements show that flies can ingest as much as 1.7× their body mass over 24 h. Besides the study of appetite, the CAFE can be used to monitor oral drug delivery. As an illustration, we used the CAFE to test the effects of dietary supplementation with two compounds, paraquat and ethanol, on food ingestion and preference. Paraquat, a prooxidant widely used in stress tests, had a strong anorexigenic effect. In contrast, in a feeding preference assay, ethanol-laced food, but not ethanol by itself, acted as an attractant. PMID:17494737

  17. Assaying Visual Memory in the Desert Locust

    Directory of Open Access Journals (Sweden)

    Senne Dillen

    2015-04-01

    Full Text Available The involvement of associative learning cues has been demonstrated in several stages of feeding and food selection. Short neuropeptide F (sNPF, an insect neuropeptide whose effects on feeding behavior have previously been well established, may be one of the factors bridging feeding and learning behavior. Recently, it was shown in Drosophila melanogaster that the targeted reduction of Drome-sNPF transcript levels significantly reduced sugar-rewarded olfactory memory. While Drosophila mainly relies on olfactory perception in its food searching behavior, locust foraging behavior is likely to be more visually orientated. Furthermore, a feeding-dependent regulation of Schgr-sNPF transcript levels has previously been observed in the optic lobes of the locust brain, suggesting a possible involvement in visual perception of food and visual associative memory in this insect species. In this study, we describe the development of a robust and reproducible assay allowing visual associative memory to be studied in the desert locust, Schistocerca gregaria. Furthermore, we performed an exploratory series of experiments, studying the role of Schgr-sNPF in this complex process.

  18. Liquid chromatographic assay for dicloxacillin in plasma.

    Science.gov (United States)

    Alderete, Oscar; González-Esquivel, Dinora F; Del Rivero, L Misael; Castro Torres, Nelly

    2004-06-15

    A simple high-performance liquid chromatographic method for the determination of dicloxacillin in plasma has been developed. The method only requires 0.5 ml of plasma, phosphate buffer solution (pH = 4.7), acidification with 0.5N hydrochloride acid and liquid extraction with dichloromethane. Posterior evaporation of organic under nitrogen steam and redissolution in mobile phase is carried out. The analysis was performed on a Spherisorb C18 (5 microm) column, using methanol -0.05 M phosphate buffer, pH = 4.7 (75:25; v/v) as mobile phase, with ultraviolet detection at 220 nm. Results showed that the assay is sensitive: 0.5 microg/ml. The response is linear in the range of 0.5 - 10 microg/ml. Maximum inter-day coefficient of variation was 12.4%. Mean extraction recovery obtained was 96.95%. Stability studies showed that the loss was not higher than 10%, samples are stable at room temperature for 6 h, at -20 Celsius for 2 months, processed samples were stable at least for 24 h and also after two freeze-thaw cycles. The method has been used to perform pharmacokinetic and bioequivalence studies in humans.

  19. Fluorometric enzymatic assay of L-arginine

    Science.gov (United States)

    Stasyuk, Nataliya; Gayda, Galina; Yepremyan, Hasmik; Stepien, Agnieszka; Gonchar, Mykhailo

    2017-01-01

    The enzymes of L-arginine (further - Arg) metabolism are promising tools for elaboration of selective methods for quantitative Arg analysis. In our study we propose an enzymatic method for Arg assay based on fluorometric monitoring of ammonia, a final product of Arg splitting by human liver arginase I (further - arginase), isolated from the recombinant yeast strain, and commercial urease. The selective analysis of ammonia (at 415 nm under excitation at 360 nm) is based on reaction with o-phthalaldehyde (OPA) in the presence of sulfite in alkali medium: these conditions permit to avoid the reaction of OPA with any amino acid. A linearity range of the fluorometric arginase-urease-OPA method is from 100 nM to 6 μМ with a limit of detection of 34 nM Arg. The method was used for the quantitative determination of Arg in the pooled sample of blood serum. The obtained results proved to be in a good correlation with the reference enzymatic method and literature data. The proposed arginase-urease-OPA method being sensitive, economical, selective and suitable for both routine and micro-volume formats, can be used in clinical diagnostics for the simultaneous determination of Arg as well as urea and ammonia in serum samples.

  20. PARALLEL ASSAY OF OXYGEN EQUILIBRIA OF HEMOGLOBIN

    Science.gov (United States)

    Lilly, Laura E.; Blinebry, Sara K.; Viscardi, Chelsea M.; Perez, Luis; Bonaventura, Joe; McMahon, Tim J.

    2013-01-01

    Methods to systematically analyze in parallel the function of multiple protein or cell samples in vivo or ex vivo (i.e. functional proteomics) in a controlled gaseous environment have thus far been limited. Here we describe an apparatus and procedure that enables, for the first time, parallel assay of oxygen equilibria in multiple samples. Using this apparatus, numerous simultaneous oxygen equilibrium curves (OECs) can be obtained under truly identical conditions from blood cell samples or purified hemoglobins (Hbs). We suggest that the ability to obtain these parallel datasets under identical conditions can be of immense value, both to biomedical researchers and clinicians who wish to monitor blood health, and to physiologists studying non-human organisms and the effects of climate change on these organisms. Parallel monitoring techniques are essential in order to better understand the functions of critical cellular proteins. The procedure can be applied to human studies, wherein an OEC can be analyzed in light of an individual’s entire genome. Here, we analyzed intraerythrocytic Hb, a protein that operates at the organism’s environmental interface and then comes into close contact with virtually all of the organism’s cells. The apparatus is theoretically scalable, and establishes a functional proteomic screen that can be correlated with genomic information on the same individuals. This new method is expected to accelerate our general understanding of protein function, an increasingly challenging objective as advances in proteomic and genomic throughput outpace the ability to study proteins’ functional properties. PMID:23827235

  1. Microrheological Coagulation Assay Exploiting Micromechanical Resonators.

    Science.gov (United States)

    Padovani, Francesco; Duffy, James; Hegner, Martin

    2017-01-03

    Rheological measurements in biological liquids yield insights into homeostasis and provide information on important molecular processes that affect fluidity. We present a fully automated cantilever-based method for highly precise and sensitive measurements of microliter sample volumes of human blood plasma coagulation (0.009 cP for viscosity range 0.5-3 cP and 0.0012 g/cm(3) for density range 0.9-1.1 g/cm(3)). Microcantilever arrays are driven by a piezoelectric element, and resonance frequencies and quality factors of sensors that change over time are evaluated. A highly accurate approximation of the hydrodynamic function is introduced that correlates resonance frequency and quality factor of cantilever beams immersed in a fluid to the viscosity and density of that fluid. The theoretical model was validated using glycerol reference solutions. We present a surface functionalization protocol that allows minimization of unspecific protein adsorption onto cantilevers. Adsorption leads to measurement distortions and incorrect estimation of the fluid parameters (viscosity and density). Two hydrophilic terminated self-assembled monolayers (SAMs) sensor surfaces are compared to a hydrophobic terminated SAM coating. As expected, the hydrophobic modified surfaces induced the highest mass adsorption and could promote conformational changes of the proteins and subsequent abnormal biological activity. Finally, the activated partial thromboplastin time (aPTT) coagulation assay was performed, and the viscosity, density, and coagulation rate of human blood plasma were measured along with the standard coagulation time. The method could extend and improve current coagulation testing.

  2. Fluorescence Polarization Assays in Small Molecule Screening

    Science.gov (United States)

    Lea, Wendy A.; Simeonov, Anton

    2011-01-01

    Importance of the field Fluorescence polarization (FP) is a homogeneous method that allows rapid and quantitative analysis of diverse molecular interactions and enzyme activities. This technique has been widely utilized in clinical and biomedical settings, including the diagnosis of certain diseases and monitoring therapeutic drug levels in body fluids. Recent developments in the field has been symbolized by the facile adoption of FP in high-throughput screening (HTS) and small molecule drug discovery of an increasing range of target classes. Areas covered in this review The article provides a brief overview on the theoretical foundation of FP, followed by updates on recent advancements in its application for various drug target classes, including G-protein coupled receptors (GPCRs), enzymes and protein-protein interactions (PPIs). The strengths and weaknesses of this method, practical considerations in assay design, novel applications, and future directions are also discussed. What the reader will gain The reader will be informed of the most recent advancements and future directions of FP application to small molecule screening. Take home message In addition to its continued utilization in high-throughput screening, FP has expanded into new disease and target areas and has been marked by increased use of labeled small molecule ligands for receptor binding studies. PMID:22328899

  3. Background Assay and Rejection in DRIFT

    CERN Document Server

    Brack, Jeff; Dorofeev, Alexei; Ezeribe, Anthony; Gauvreau, Jean-Luc; Gold, Michael; Harton, John; Lafler, Randy; Lauer, Robert; Lee, Eric R; Loomba, Dinesh; Matthews, John; Miller, Eric H; Monte, Alissa; Murphy, Alex; Paling, Sean; Phan, Nguyen; Sadler, Steve; Scarff, Andrew; Snowden-Ifft, Daniel; Spooner, Neil; Telfer, Sam; Walker, Daniel; Williams, Matt; Yuriev, Leonid

    2014-01-01

    The DRIFT-IId dark matter detector is a m$^3$-scale low-pressure TPC with directional sensitivity to WIMP-induced nuclear recoils. Its primary backgrounds were due to alpha decays from contamination on the central cathode. Efforts to reduce these backgrounds led to replacing the 20 \\mu m wire central cathode with one constructed from 0.9 \\mu m aluminized mylar, which is almost totally transparent to alpha particles. Detailed modeling of the nature and origin of the remaining backgrounds led to an in-situ, ppt-sensitive assay of alpha decay backgrounds from the central cathode. This led to further improvements in the thin-film cathode resulting in over 2 orders of magnitude reduction in backgrounds compared to the wire cathode. Finally, the addition of O$_2$ to CS$_2$ gas was found to produce multiple species of electronegative charge carriers, providing a method to determine the absolute position of nuclear recoils and reject all known remaining backgrounds while retaining a high efficiency for nuclear recoil...

  4. Mouse lung adhesion assay for Bordetella pertussis

    Energy Technology Data Exchange (ETDEWEB)

    Burns, K.A.; Freer, J.H. (Department of Microbiology, Alexander Stone Building, Bearsden, Glasgow, Scotland)

    1982-03-01

    The ability of Bordetella pertussis to adhere to cell surfaces has been demonstrated by adhesion to tissue culture cells and adhesion to chicken, hamster or rabbit trachea in organ culture. In this report a mouse lung assay for adhesion is described and the results obtained using two virulent strains of B. pertussis and their avirulent counterparts. These were a C modulation of one of the original virulent strains and a phase IV variant of the other virulent strain. Organisms were radiolabelled by adding 1 ..mu..Ci (37 K Bq) of (/sup 14/C)glutamic acid per 10 ml of culture medium before inoculation and incubation for 5 days. The lungs were washed by perfusion in situ with at least two volumes (1 ml) of sterile 1% (w/v) casamino acids. The percentage of the inoculated organisms retained in the lungs was determined, after removal of the lungs, by one of the following two methods: viable count or radioactive count. Results for both methods were expressed as the percentage of the inoculum retained in the lungs plus or minus one standard deviation.

  5. Rho family and Rap GTPase activation assays.

    Science.gov (United States)

    Jennings, Richard T; Knaus, Ulla G

    2014-01-01

    The detection of Ras superfamily GTPase activity in innate immune cells is important when studying signaling events elicited by various ligands and cellular processes. The development of high-affinity probes detecting the activated, GTP-bound form of small GTPases has significantly enhanced our understanding of initiation and termination of GTPase-regulated signaling pathways. These probes are created by fusing a high-affinity GTPase-binding domain derived from a specific downstream effector protein to glutathione S-transferase (GST). Such domains bind preferentially to the GTP-bound form of the upstream Rho or Ras GTPase. Coupling these probes to beads enables extraction of the complex and subsequent quantification of the active GTP-binding protein by immunoblotting. Although effector domains that discriminate efficiently between GDP- and GTP-bound states and highly specific antibodies are not yet available for every small GTPase, analysis of certain members of the Rho and Ras GTPase family is now routinely performed. Here, we describe affinity-based pulldown assays for detection of Rho GTPase (Rac1/2, Cdc42, RhoA/B) and Rap1/2 activity in stimulated neutrophils or macrophages.

  6. Establishment of indirect immunofluorescence assay for rotavirus.

    Science.gov (United States)

    Tao, J; Zhang, J; Liu, X; Jin, H; Jiang, C; Yin, Y

    2016-03-01

    Rotavirus infection is the most frequent cause of infantile gastroenteritis worldwide and a significant cause of death in infants and young children, following severe diarrhea and dehydration. Rotavirus vaccines are considered the most effective way to prevent rotavirus infections. In the process of developing rotavirus vaccines, it is crucial to establish a reliable and standardized method to determine vaccine titer. In this study, we developed an indirect immunofluorescence assay (IFA) to determine the infectious titer of Lanzhou lamb rotavirus (LLR) vaccine grown in MA104 cells. The activating concentration of trypsin was 1 µg/ml for healthy monolayers of MA104 cells at 100% confluence. After incubation for 18 hr, a rabbit anti-SA11 polyclonal antibody, diluted at 1:800 in PBS, was added to all wells, followed by an Alexa-488-conjugated secondary antibody diluted at 1:500 in PBS. Cells were examined with a fluorescence microscope. Our results show that IFA was more reproducible, more sensitive, simpler, and more rapid than the log 50% cell culture infectious dose-ELISA (lgCCID50-ELISA) in measuring the rotavirus vaccines. IFA provided a reliable basis for the qualitative and quantitative analysis of rotavirus, and the certification of rotavirus vaccine production.

  7. A comprehensive company database analysis of biological assay variability.

    Science.gov (United States)

    Kramer, Christian; Dahl, Göran; Tyrchan, Christian; Ulander, Johan

    2016-08-01

    Analysis of data from various compounds measured in diverse biological assays is a central part of drug discovery research projects. However, no systematic overview of the variability in biological assays has been published and judgments on assay quality and robustness of data are often based on personal belief and experience within the drug discovery community. To address this we performed a reproducibility analysis of all biological assays at AstraZeneca between 2005 and 2014. We found an average experimental uncertainty of less than a twofold difference and no technologies or assay types had higher variability than others. This work suggests that robust data can be obtained from the most commonly applied biological assays.

  8. Optimization of cell motility evaluation in scratch assay

    Directory of Open Access Journals (Sweden)

    Gotsulyak N. Ya.

    2014-05-01

    Full Text Available A scratch test is one of the most popular methods of classical cell migration assay in a monolayer culture. At the same time, the scratch assay has some disadvantages that can be easily corrected. Aim. Optimization the existent scratch assay on the base of detection of scratch wound surface area and the length of the field of observation which is more objective and less time consuming. Methods. Scratch assay. Results. The modification of scratch assay enables to perform measurement more accurately and rapidly. This approach is more simple and eliminates the main disadvantages of the classical method. Conclusions. The procedure of scratch wound width measurement calculated on the base of detection of cell free area and the length of the field of observation is more effective than the classical wound healing assay. It will be useful for the estimation of cell migration dynamics in monolayer culture for a wide range of live cell based experiments.

  9. Ecotoxicological Assessment of Aquatic Genotoxicity Using the Comet Assay

    Directory of Open Access Journals (Sweden)

    KHUSNUL YAQIN

    2006-09-01

    Full Text Available Comet assay is a novel biological analysis, which is a sensitive, flexible, simple, rapid, and inexpensive method to assess aquatic genotoxicant. Since Singh and co-workers developed the method in 1988, its use has increased exponentially in various fields. This review discourses on the application of this assay in aquatic ecosystems. Various types of cells from various aquatic organisms have been tested by various genotoxicant both direct- and indirect-acting using the comet assay. The applications of this assay suggest that it is a useful assay to assess aquatic genotoxicants. However, there are some factors, which should be taken into account when using this assay as aquatic ecotoxicological assessment device such as inter-animal and cell variability.

  10. Establishment of an Enzyme Linked Immunosorbent Assay for Neonatal Thyrotropin

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A sensitive and specific ELISA for neonatal thyrotropin(Neonatal TSH) is established with using twoanti-TSH monoclonal antibody. One of them is coated on the microtiter plate, the other is conjugate ofbiotin. The label is horseradish peroxidase(HRP) conjugate of streptavidin. TMB-H2O2 solution is used asthe substrate of HRP.The sensitivity of the assay is 0.5 mIU/L, the intra-assay CVs and the intre-assay

  11. Validation of Laboratory-Developed Molecular Assays for Infectious Diseases

    OpenAIRE

    Burd, Eileen M.

    2010-01-01

    Summary: Molecular technology has changed the way that clinical laboratories diagnose and manage many infectious diseases. Excellent sensitivity, specificity, and speed have made molecular assays an attractive alternative to culture or enzyme immunoassay methods. Many molecular assays are commercially available and FDA approved. Others, especially those that test for less common analytes, are often laboratory developed. Laboratories also often modify FDA-approved assays to include different e...

  12. An ultrasensitive rapid immunocytotoxicity assay for detecting Clostridium difficile toxins

    Science.gov (United States)

    He, Xiangyun; Wang, Jufang; Steele, Jennifer; Sun, Xingmin; Nie, Weijia; Tzipori, Saul; Feng, Hanping

    2009-01-01

    We describe a novel ultrasensitive cell-based immunocytotoxicity assay for detecting less then 1 pg/ml of Clostridium difficile toxins in porcine clinical samples. The assay is simple to perform with a turnaround time of approximately 3 hours and capable of detecting less then 1 pg/ml of toxin A. Using this assay, we were able to detect the presence of C. difficile toxins in the fecal and serum specimens of experimentally infected piglets. PMID:19393695

  13. The Spheroplast Lysis Assay for Yeast in Microtiter Plate Format

    Science.gov (United States)

    Ovalle, Rafael; Spencer, Moyah; Thiwanont, Monthiwa; Lipke, Peter N.

    1999-01-01

    A yeast lysis assay in the microtiter plate format improved precision and throughput and led to an improved algorithm for estimating lag time. The assay reproducibly revealed differences of 10% or greater in the maximal lysis rate and 50% or greater in the lag time. Clonal differences were determined to be the major source of variation. Microtiter-based assays should be useful for screening for drug susceptibility and for analyzing mutant phenotypes. PMID:10427014

  14. BROMATOMATRIC ASSAY OF GATIFLOXACIN IN PHARMACEUTICALS

    Directory of Open Access Journals (Sweden)

    KALSANG THARPA

    2008-09-01

    Full Text Available Three new, simple, and cost-effective visible spectrophotometric methods are proposed for determination of gatifloxacin (GTF using bromate-bromide mixture, and three dyes, methyl orange, indigocarmine and thymol blue, as reagents.The methods engross the addition of a known excess of bromate-bromide mixture to GTF in hydrochloric acid medium followed by determination of residual bromine by reacting with a fixed amount of either methyl orange andmeasuring the absorbance at 520 nm (method A or indigo carmine and measuring the absorbance at 610 nm (method B or thymol blue and measuring the absorbance at 550 nm (method C. In all the methods, the amount of brominereacted corresponds to the amount of GTF, and the absorbance is found to increase linearly with the concentration of GTF. Under the optimum conditions, GTF could be assayed in the concentration range 0.25-1.5, 0.5-6.0, and 0.5-10μg/mL by method A, method B and method C, respectively. The apparent molar absorptivities are calculated to be 1.6x105, 4.0x104 and 3.2x104 L mol-1 cm-1 for the method A, method B and method C, respectively, and the corresponding Sandell sensitivity values are 0.0025, 0.010 and 0.012 μg/cm2. The intra-day and inter-day precision, and the accuracy of the methods were evaluated as per the current ICH guidelines. The methods were successfully applied to the determination of GTF in pharmaceutical preparations without the interference from any of the pharmaceutical adjuvants.

  15. Inferring Protein Associations Using Protein Pulldown Assays

    Energy Technology Data Exchange (ETDEWEB)

    Sharp, Julia L.; Anderson, Kevin K.; Daly, Don S.; Auberry, Deanna L.; Borkowski, John J.; Cannon, William R.

    2007-02-01

    Background: One method to infer protein-protein associations is through a “bait-prey pulldown” assay using a protein affinity agent and an LC-MS (liquid chromatography-mass spectrometry)-based protein identification method. False positive and negative protein identifications are not uncommon, however, leading to incorrect inferences. Methods: A pulldown experiment generates a protein association matrix wherein each column represents a sample from one bait protein, each row represents one prey protein and each cell contains a presence/absence association indicator. Our method evaluates the presence/absence pattern across a prey protein (row) with a Likelihood Ratio Test (LRT), computing its p-value with simulated LRT test statistic distributions after a check with simulated binomial random variates disqualified the large sample 2 test. A pulldown experiment often involves hundreds of tests so we apply the false discovery rate method to control the false positive rate. Based on the p-value, each prey protein is assigned a category (specific association, non-specific association, or not associated) and appraised with respect to the pulldown experiment’s goal and design. The method is illustrated using a pulldown experiment investigating the protein complexes of Shewanella oneidensis MR-1. Results: The Monte Carlo simulated LRT p-values objectively reveal specific and ubiquitous prey, as well as potential systematic errors. The example analysis shows the results to be biologically sensible and more realistic than the ad hoc screening methods previously utilized. Conclusions: The method presented appears to be informative for screening for protein-protein associations.

  16. Comparative investigation of various cellulase assay procedures

    Energy Technology Data Exchange (ETDEWEB)

    Canevascini, G.; Gattlen, C.

    1981-07-01

    The cellulolytic activity of crude enzyme preparations from different cellulolytic fungi (namely Trichoderma viride, Trichoderma koningii, Fusarium solani, Sporotrichum pulverulentum, Sporotrichum thermophile) was assayed comparatively with several common analytical procedures described in the literature. The investigation was carried out with the objective of evaluating, with raw culture filtrates, the different cellulase tests in relation to their specificity for endo- and exo-cellulase action as well as to allow comparisons to be made between results from different research groups using different methods. 1) Cellulase activity was tested viscometrically as well as chemically (determination of reducing end groups) with different carboxymethylcelluloses as substrates. Essentially constant ratios between both kinds of activities were obtained, indicating that they are directly related. By estimating cellulase activity with insoluble cellulosic substrates no direct relationship could be established with the above-described activities except in the case where the cellulose was amorphous. The ratio profile between activities thus obtained and endo-cellulase activities determined viscometrically shows that some enzyme preparations (such as those from both Trichoderma sp.) are clearly more active than others against crystalline cellulose reflecting quantitative differences in enzyme composition. Nevertheless, for a biological understanding of cellulolysis, analytical procedures using crystalline celluloses are not adequate for specifically monitoring exo-cellulase activity in crude enzyme solutions for essentially two reasons: a) they are not sufficiently sensitive to detect small changes in enzyme activity during the early phase of growth, and b) exo-cellulase activity in crude enzyme solutions also depends on the endo-cellulase activity present. (Refs. 39).

  17. Robust versatile tyrosine kinase assay for HTS in drug discovery

    Science.gov (United States)

    Deshpande, Sudhir S.; Mineyev, I.; Owicki, John C.

    1999-04-01

    A fluorescence polarization assay was developed as an alternative to the radiolabeled SPA assays currently used to monitor the activity of tyrosine kinases in drug discovery. The assay can be used with enzymes having substrate specificity similar to that of the insulin receptor, the EGF receptor and the Src kinase receptor enzymes. The assay is easy to configure in 96, 384 and 1536-well microplates in assay volumes ranging from (mu) L with minimal efforts. The reconstituted reagents are stable for up to 24 hr at ambient temperatures, thereby minimizing the need for replenishing the stock solutions during the course of a high-throughput screen. Because of the stability and equilibrium kinetics, the assay allows the user the luxury of scheduling the reading of plates any time up to 24 hr after the completion of the assay without substantial deterioration in the assay signal. The antibody and the tracer solutions can also be premixed and added as a preformed complex in a single step. The performance of the assay with the insulin receptor kinase is described. In addition, given the diversity of the substrates used in measuring the activity of different tyrosine kinases, LJL's on-going efforts to provide different antibodies of wide ranging specificity and sensitivity are described.

  18. Click Chemistry-Mediated Nanosensors for Biochemical Assays

    National Research Council Canada - National Science Library

    Chen, Yiping; Xianyu, Yunlei; Wu, Jing; Yin, Binfeng; Jiang, Xingyu

    2016-01-01

    Click chemistry combined with functional nanoparticles have drawn increasing attention in biochemical assays because they are promising in developing biosensors with effective signal transformation...

  19. A Multi-detection Assay for Malaria Transmitting Mosquitoes

    Science.gov (United States)

    Lee, Yoosook; Weakley, Allison M.; Nieman, Catelyn C.; Malvick, Julia; Lanzaro, Gregory C.

    2015-01-01

    The Anopheles gambiae species complex includes the major malaria transmitting mosquitoes in Africa. Because these species are of such medical importance, several traits are typically characterized using molecular assays to aid in epidemiological studies. These traits include species identification, insecticide resistance, parasite infection status, and host preference. Since populations of the Anopheles gambiae complex are morphologically indistinguishable, a polymerase chain reaction (PCR) is traditionally used to identify species. Once the species is known, several downstream assays are routinely performed to elucidate further characteristics. For instance, mutations known as KDR in a para gene confer resistance against DDT and pyrethroid insecticides. Additionally, enzyme-linked immunosorbent assays (ELISAs) or Plasmodium parasite DNA detection PCR assays are used to detect parasites present in mosquito tissues. Lastly, a combination of PCR and restriction enzyme digests can be used to elucidate host preference (e.g., human vs. animal blood) by screening the mosquito bloodmeal for host-specific DNA. We have developed a multi-detection assay (MDA) that combines all of the aforementioned assays into a single multiplex reaction genotyping 33SNPs for 96 or 384 samples at a time. Because the MDA includes multiple markers for species, Plasmodium detection, and host blood identification, the likelihood of generating false positives or negatives is greatly reduced from previous assays that include only one marker per trait. This robust and simple assay can detect these key mosquito traits cost-effectively and in a fraction of the time of existing assays. PMID:25867057

  20. Modeling error in experimental assays using the bootstrap principle: understanding discrepancies between assays using different dispensing technologies

    Science.gov (United States)

    Hanson, Sonya M.; Ekins, Sean; Chodera, John D.

    2015-12-01

    All experimental assay data contains error, but the magnitude, type, and primary origin of this error is often not obvious. Here, we describe a simple set of assay modeling techniques based on the bootstrap principle that allow sources of error and bias to be simulated and propagated into assay results. We demonstrate how deceptively simple operations—such as the creation of a dilution series with a robotic liquid handler—can significantly amplify imprecision and even contribute substantially to bias. To illustrate these techniques, we review an example of how the choice of dispensing technology can impact assay measurements, and show how large contributions to discrepancies between assays can be easily understood and potentially corrected for. These simple modeling techniques—illustrated with an accompanying IPython notebook—can allow modelers to understand the expected error and bias in experimental datasets, and even help experimentalists design assays to more effectively reach accuracy and imprecision goals.

  1. Modeling error in experimental assays using the bootstrap principle: understanding discrepancies between assays using different dispensing technologies.

    Science.gov (United States)

    Hanson, Sonya M; Ekins, Sean; Chodera, John D

    2015-12-01

    All experimental assay data contains error, but the magnitude, type, and primary origin of this error is often not obvious. Here, we describe a simple set of assay modeling techniques based on the bootstrap principle that allow sources of error and bias to be simulated and propagated into assay results. We demonstrate how deceptively simple operations--such as the creation of a dilution series with a robotic liquid handler--can significantly amplify imprecision and even contribute substantially to bias. To illustrate these techniques, we review an example of how the choice of dispensing technology can impact assay measurements, and show how large contributions to discrepancies between assays can be easily understood and potentially corrected for. These simple modeling techniques--illustrated with an accompanying IPython notebook--can allow modelers to understand the expected error and bias in experimental datasets, and even help experimentalists design assays to more effectively reach accuracy and imprecision goals.

  2. Assay optimization for molecular detection of Zika virus

    Science.gov (United States)

    Corman, Victor M; Rasche, Andrea; Baronti, Cecile; Aldabbagh, Souhaib; Cadar, Daniel; Reusken, Chantal BEM; Pas, Suzan D; Goorhuis, Abraham; Schinkel, Janke; Molenkamp, Richard; Kümmerer, Beate M; Bleicker, Tobias; Brünink, Sebastian; Eschbach-Bludau, Monika; Eis-Hübinger, Anna M; Koopmans, Marion P; Schmidt-Chanasit, Jonas; Grobusch, Martin P; de Lamballerie, Xavier; Drosten, Christian

    2016-01-01

    Abstract Objective To examine the diagnostic performance of real-time reverse transcription (RT)-polymerase chain reaction (PCR) assays for Zika virus detection. Methods We compared seven published real-time RT–PCR assays and two new assays that we have developed. To determine the analytical sensitivity of each assay, we constructed a synthetic universal control ribonucleic acid (uncRNA) containing all of the assays’ target regions on one RNA strand and spiked human blood or urine with known quantities of African or Asian Zika virus strains. Viral loads in 33 samples from Zika virus-infected patients were determined by using one of the new assays. Findings Oligonucleotides of the published real-time RT–PCR assays, showed up to 10 potential mismatches with the Asian lineage causing the current outbreak, compared with 0 to 4 mismatches for the new assays. The 95% lower detection limit of the seven most sensitive assays ranged from 2.1 to 12.1 uncRNA copies/reaction. Two assays had lower sensitivities of 17.0 and 1373.3 uncRNA copies/reaction and showed a similar sensitivity when using spiked samples. The mean viral loads in samples from Zika virus-infected patients were 5 × 104 RNA copies/mL of blood and 2 × 104 RNA copies/mL of urine. Conclusion We provide reagents and updated protocols for Zika virus detection suitable for the current outbreak strains. Some published assays might be unsuitable for Zika virus detection, due to the limited sensitivity and potential incompatibility with some strains. Viral concentrations in the clinical samples were close to the technical detection limit, suggesting that the use of insensitive assays will cause false-negative results. PMID:27994281

  3. ToxCast HTS Assay Development and Retrofitting: Strategies ...

    Science.gov (United States)

    A presentation to EC JRC partners on new ToxCast HTS assay methods and strategies to address current limitations to HTS methods Slide presentation to EC JRC partners on new ToxCast HTS assay methods and strategies to address current limitations to HTS methods.

  4. Nanoparticle-assay marker interaction: effects on nanotoxicity assessment

    Science.gov (United States)

    Zhao, Xinxin; Xiong, Sijing; Huang, Liwen Charlotte; Ng, Kee Woei; Loo, Say Chye Joachim

    2015-01-01

    Protein-based cytotoxicity assays such as lactate dehydrogenase (LDH) and tumor necrosis factor-alpha (TNF-α) are commonly used in cytotoxic evaluation of nanoparticles (NPs) despite numerous reports on possible interactions with protein markers in these assays that can confound the results obtained. In this study, conventional cytotoxicity assays where assay markers may (LDH and TNF- α) or may not (PicoGreen and WST-8) come into contact with NPs were used to evaluate the cytotoxicity of NPs. The findings revealed selective interactions between negatively charged protein assay markers (LDH and TNF- α) and positively charged ZnO NPs under abiotic conditions. The adsorption and interaction with these protein assay markers were strongly influenced by surface charge, concentration, and specific surface area of the NPs, thereby resulting in less than accurate cytotoxic measurements, as observed from actual cell viability measurements. An improved protocol for LDH assay was, therefore, proposed and validated by eliminating any effects associated with protein-particle interactions. In view of this, additional measures and precautions should be taken when evaluating cytotoxicity of NPs with standard protein-based assays, particularly when they are of opposite charges.

  5. Engineering luciferase enzymes and substrates for novel assay capabilities

    Science.gov (United States)

    Wood, Keith V.

    2004-06-01

    In the development of HTS as a central paradigm of drug discovery, fluorescent reporter molecules have generally been adopted as the favored signal transducer. Nevertheless, luminescence has maintained a prominent position among certain methodologies, most notably genetic reporters. Recently, there has been growing partiality for luminescent assays across a broader range of applications due to their sensitivity, extensive linearity, and robustness to library compounds and complex biological samples. This trend has been fostered by development several new assay designs for diverse targets such as kinases, cytochrome p450's, proteases, apoptosis, and cytotoxicity. This review addresses recent progress made in the use of bioluminescent assays for drug discovery, highlighting new detection capabilities brought about by engineering luciferase enzymes and substrates. In reporter gene applications, modified luciferases have provided greatly improved expression efficiency in mammalian cells, improved responsiveness to changes of transcriptional rate, and increased the magnitude of the reporter response. Highly stabilized luciferase mutants have enabled new assays strategies for high-throughput screening based on detection of ATP and luciferin. Assays based on ATP support rapid analysis of cell metabolism and enzymatic processes coupled to ATP hydrolysis. Although luciferin is found natively only in luminous beetles, coupled assays have been designed using modified forms of luciferin requiring the action of second enzyme to yield luminescence. Due to the very low inherent background and protection of the photon-emitter afforded by the enzyme, bioluminescent assays often outperform the analogous fluorescent assays for analyses performed in multiwell plates.

  6. Colorimetric micro-assay for accelerated screening of mould inhibitors

    Science.gov (United States)

    Carol A. Clausen; Vina W. Yang

    2013-01-01

    Since current standard laboratory methods are time-consuming macro-assays that rely on subjective visual ratings of mould growth, rapid and quantitative laboratory methods are needed to screen potential mould inhibitors for use in and on cellulose-based products. A colorimetric micro-assay has been developed that uses XTT tetrazolium salt to enzymatically assess...

  7. Radioimmunoprecipitation polyethylene glycol assay for circulating Entamoeba histolytica antigens

    Energy Technology Data Exchange (ETDEWEB)

    Pillai, S.; Mohimen, A.; Mehra, S. (Calcutta Medical Research Inst., Calcutta (India). Kothari Centre of Gastroenterology)

    1982-12-17

    An assay capable of detecting circulating Entamoeba histolytica antigens in amoebiasis is described. This assay utilised a radiolabelled affinity purified rabbit anti-E. histolytica antibody that had been depleted of antibodies that cross-react with human serum proteins, and a polyethylene glycol precipitation step.

  8. Microfluidic assay without blocking for rapid HIV screening and confirmation.

    Science.gov (United States)

    Song, Lusheng; Zhang, Yi; Wang, Wenjun; Ma, Liying; Liu, Yong; Hao, Yanlin; Shao, Yiming; Zhang, Wei; Jiang, Xingyu

    2012-08-01

    The essential step for HIV spreading limitation is the screening tests. However, there are multiple disadvantages in current screening assays which need further confirmation test. Herein we developed a rapid HIV assay combining screening and confirmation test by using the microfluidic network assay. Meanwhile, the assay is accelerated by bypassing the step of blocking. We call this method as microfluidic assay without blocking (MAWB). Both the limit of detection and reagent incubation time of MAWB are determined by screening of one model protein pair: ovalbumin and its antibody. The assay time is accelerated about 25% while the limit of detection (LOD) is well kept. Formatting the method in for both HIV screening (testing 8 HIV-related samples) and confirmation (assaying 6 kinds of HIV antibodies of each sample) within 30 min was successful. Fast HIV screening and confirmation of 20 plasma samples were also demonstrated by this method. MAWB improved the assay speed while keeping the LOD of conventional ELISA. Meanwhile, both the accuracy and throughput of MAWB were well improved, which made it an excellent candidate for a quick HIV test for both screening and confirmation. Methods like this one will find wide applications in clinical diagnosis and biochemical analysis based on the interactions between pairs of molecules.

  9. The glucose oxidase-peroxidase assay for glucose

    Science.gov (United States)

    The glucose oxidase-peroxidase assay for glucose has served as a very specific, sensitive, and repeatable assay for detection of glucose in biological samples. It has been used successfully for analysis of glucose in samples from blood and urine, to analysis of glucose released from starch or glycog...

  10. A Simple Endpoint Assay for Starch-Degrading Enzymes.

    Science.gov (United States)

    Kroen, William K.

    1998-01-01

    Since many of the important energy-transferring pathways involve synthesis or degradation of biological macromolecules, observations of the enzymes responsible for starch breakdown provide a useful case study. Provides a short, one-step assay for the enzymes amylase and amyloglucosidase. Topics covered include goals, preparation, assay procedure,…

  11. 21 CFR 864.7455 - Fetal hemoglobin assay.

    Science.gov (United States)

    2010-04-01

    ... hemoglobin present. The assay may be used to detect fetal red cells in the maternal circulation or to detect... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Fetal hemoglobin assay. 864.7455 Section 864.7455...) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7455 Fetal...

  12. Exposure-based validation list for developmental toxicity screening assays

    NARCIS (Netherlands)

    Daston, George P.; Beyer, Bruce K.; Carney, Edward W.; Chapin, Robert E.; Friedman, Jan M.; Piersma, Aldert H.; Rogers, John M.; Scialli, Anthony R.

    2014-01-01

    Validation of alternative assays requires comparison of the responses to toxicants in the alternative assay with in vivo responses. Chemicals have been classified as "positive" or "negative" in vivo, despite the fact that developmental toxicity is conditional on magnitude of exposure. We developed a

  13. Nanoparticle-assay marker interaction: effects on nanotoxicity assessment

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Xinxin; Xiong, Sijing; Huang, Liwen Charlotte; Ng, Kee Woei, E-mail: kwng@ntu.edu.sg; Loo, Say Chye Joachim, E-mail: joachimloo@ntu.edu.sg [Nanyang Technological University, School of Materials Science and Engineering (Singapore)

    2015-01-15

    Protein-based cytotoxicity assays such as lactate dehydrogenase (LDH) and tumor necrosis factor-alpha (TNF-α) are commonly used in cytotoxic evaluation of nanoparticles (NPs) despite numerous reports on possible interactions with protein markers in these assays that can confound the results obtained. In this study, conventional cytotoxicity assays where assay markers may (LDH and TNF- α) or may not (PicoGreen and WST-8) come into contact with NPs were used to evaluate the cytotoxicity of NPs. The findings revealed selective interactions between negatively charged protein assay markers (LDH and TNF- α) and positively charged ZnO NPs under abiotic conditions. The adsorption and interaction with these protein assay markers were strongly influenced by surface charge, concentration, and specific surface area of the NPs, thereby resulting in less than accurate cytotoxic measurements, as observed from actual cell viability measurements. An improved protocol for LDH assay was, therefore, proposed and validated by eliminating any effects associated with protein–particle interactions. In view of this, additional measures and precautions should be taken when evaluating cytotoxicity of NPs with standard protein-based assays, particularly when they are of opposite charges.

  14. Bioluminescence-Sensing Assay for Microbial Growth Recognition

    Directory of Open Access Journals (Sweden)

    Heba Ramadan Eed

    2016-01-01

    Full Text Available The conventional methods for microbial viability quantification require cultivation and are laborious. There is consequently a widespread need for cultivation-free methods. The adenosine triphosphate (ATP bioluminescence-sensing assay is considered an extremely effective biosensor; hence ATP is the energy currency of all living microbes and can be used as a rapid indicator of microbial viability. We developed an ATP bioluminescence-sensing assay to detect microbial viability. A bioluminescent recombinant E. coli strain was used with luciferase extracted from transformed bacteria. Results showed that there is a direct correlation between the bioluminescence intensity of the ATP bioluminescence-sensing assay and the microbial viability. Bacterial counts from food samples were detected using the developed sensing assay and validated by the traditional plate-counting method. Compared with the plate-counting method, ATP bioluminescence-sensing assay is a more rapid and efficient approach for detecting microbial viability.

  15. Comparison of immunoturbidimetric and immunonephelometric assays for specific proteins.

    Science.gov (United States)

    Mali, Bahera; Armbruster, David; Serediak, Ernie; Ottenbreit, Tammy

    2009-10-01

    Immunoturbidimetric assays for specific proteins are available on "open system" clinical chemistry analyzers. The analytical performance of nine immunoturbidimetric specific protein assays (C3, C4, CRP, Haptoglobin, IgA, IgG, IgM, RF, and Transferrin) was compared to immunonephelometry. Testing was performed on the Abbott ARCHITECT ci8200 and the Dade Behring BNII nephelometer and evaluated for precision, linearity, limit of detection, prozone phenomenon, method comparison, workflow, and proficiency testing survey comparison. Immunoturbidimetric assays performance was satisfactory for total precision, linearity, limit of detection and the prozone effect was not observed. Method comparison was acceptable for the immunoglobulins, CRP and transferrin but less favorable for the other assays, likely due to methodology and antibody specificity differences. Immunourbidimetric specific protein assays allow for efficient test consolidation on a general purpose clinical chemistry analyzer.

  16. Development and Validation of a HPV-32 Specific PCR Assay

    Directory of Open Access Journals (Sweden)

    Leigh Janet

    2009-06-01

    Full Text Available Abstract Background Human Papillomavirus-32 (HPV-32 has traditionally been associated with focal-epithelial-hyperplasia (FEH. It is also present in 58% of oral warts of HIV-positive individuals whose prevalence is increasing. Current methods for the detection of HPV-32 are labor-intensive and insensitive so the goal of this work was to develop a highly sensitive and easy to use specific polymerase chain reaction (PCR assay. Materials and methods An HPV-32 L1 specific PCR assay was developed and optimized. The sensitivity and specificity was compared to previous assays utilized for detection (PGMY and MY09/11 PCR with dot blot hybridization using cloned HPV-32 L1, the closely related HPV-42 L1 as well as clinical samples (oral swabs and fluids from 89 HIV-positive subjects. Results The HPV-32 specific PCR assay showed improved sensitivity to 5 copies of HPV-32 as compared to the PGMY PCR, MY09/11 PCR and dot blot which had a limit of detection of approximately 3,000 copies. Using the HPV-32 dot blot hybridization assay as the gold standard, the HPV-32 specific PCR assay has a sensitivity of 95.8% and 88.9% by sample and subject, respectively, and specificity was 87.8% and 58.8% by sample and subject, respectively. The low sensitivity is due to the HPV-32 specific PCR assays ability to detect more HPV-32 positive samples and may be the new gold standard. Conclusion Due to the ease, sensitivity, and specificity the HPV-32 specific PCR assay is superior to previous assays and is ideal for detection of HPV-32 in large cohorts. This assay provides an excellent tool to study the natural history of HPV-32 infection and the development of oral warts.

  17. Challenges and approaches to conducting and interpreting the amphibian metamorphosis assay and the fish short-term reproduction assay.

    Science.gov (United States)

    Coady, Katherine Kemler; Lehman, Christine Marie; Currie, Rebecca J; Marino, Troy Alan

    2014-02-01

    The amphibian metamorphosis assay (AMA) and the fish short-term reproduction assay (FSTRA) are screening assays designed to detect potential endocrine activity of a test substance. These assays are included in a battery of assays in Tier 1 of U.S. Environmental Protection Agency's Endocrine Disruptor Screening Program. Based on our laboratory's experience with these two assays, we have noted several challenges in the conduct and interpretation of the AMA and FSTRA, including, but not limited to, diseased/parasitized test organisms, failure to meet some guideline performance criteria, and issues selecting and maintaining test concentrations. Various approaches are described for addressing the challenges associated with both the conduct and interpretation of these assays. Historical control data for both the AMA and FSTRA are presented to further understand background occurrences of histopathological phenomena and variability associated with the measured endpoints in these assays. In the historical control database for the AMA, wet weight on day 7 was the most variable endpoint (coefficient of variation = 26%), while developmental stage on day 21 was least variable (coefficient of variation = 0.47%). In the FSTRA, vitellogenin concentrations were the most variable endpoint (coefficient of variation = 47-84%), while fertility was the least variable endpoint (coefficient of variation = 1.5%) among historical controls.

  18. Assessment and reduction of comet assay variation in relation to DNA damage: studies from the European Comet Assay Validation Group

    DEFF Research Database (Denmark)

    Møller, Peter; Möller, Lennart; Godschalk, Roger W L

    2010-01-01

    The alkaline single cell gel electrophoresis (comet) assay has become a widely used method for the detection of DNA damage and repair in cells and tissues. Still, it has been difficult to compare results from different investigators because of differences in assay conditions and because the data ...

  19. Comet assay, cloning assay, and light and electron microscopy on one preselected cell

    Science.gov (United States)

    Koenig, Karsten; Oehring, Hartmut; Halbhuber, Karl-Juergen; Fiedler, Ursula; Bauer, Eckhard; Greulich, Karl-Otto

    1998-01-01

    In order to perform long-term studies up to one week on a preselected single cell after micromanipulation (e.g. UVA and NIR microbeam exposure) in comparison with non-treated neighbor cells (control cells) we applied a variety of single cell diagnostic techniques and developed a special comet assay for single preselected cells. For that purpose adherent cells were grown in low concentrations and maintained in special sterile centimeter-sized glass cell chambers. After preselection, a single cell was marked by means of diamond-produced circles on the outer cell chamber window. During exposure to microbeams, NADH-attributed autofluorescence of the chosen cell was detected by fluorescence imaging and spectroscopy. In addition, cell morphology was video-monitored (formation of pseudopodia, membrane blebbing,...). Maintaining the microchamber in the incubator, the irradiated cell was examined 24 h later for cell division (clone formation) and modifications in autofluorescence and morphology (including daughter cells). In the case that no division occurred the vitality of the light-exposed cell and of the control cells were probed by intranuclear propidium iodide accumulation. After fixation, either electron microscopy or single cell gel electrophoresis (comet assay) was performed. To monitor comet formation indicating photoinduced DNA damage in the preselected single cell in comparison with the non-exposed neighbor cells the chamber was filled with low-melting gel and lysis solution and exposed to an electric field. In contrast to the conventional comet assay, where only randomly chosen cells of a suspension are investigated, the novel optimized electrophoresis technique should enhance the possibilities of DNA damage detection to a true single (preselected) cell level. The single cell techniques applied to UVA microexposed Chinese hamster ovary cells (364 nm, 1 mW, 3.5 W/cm2) revealed significant cell damage for J/cm2 fluences such as modifications of intracellular

  20. The line blot assay: problems with titrating first and second antibodies for Western blot and immunohistochemistry assays?

    Science.gov (United States)

    Rojas-Espinosa, O; Silva-Miranda, M; Wek-Rodriguez, K; Arce-Paredes, P

    2006-01-01

    We describe a technique designed to assess the optimal dilution of primary and secondary antibodies, to be used in Western blot, dot blot, the multi-antigen print immunoassay (MAPIA) and immunohistochemistry assays. The method that we call "line blot" is not an alternative but a practical, complementary tool for the above techniques that assures definitive results are obtained from single assays, so there is no need to repeat the assay. As with most immunoenzymatic assays, the line blot assay is very sensitive, allowing the detection of absolute amounts of antigen as low as 2.5 ng in the 0.5 cm-long segment line (see Results), depending on the strength of the secondary, enzyme-labelled antibody.

  1. [Role of ultrasensitive TSH assay and ultrarapid total thyroxine assay in the thyroid studies].

    Science.gov (United States)

    Fragu, P; Comoy, E; Noël, M; Lounis, M; Patois, E; Parmentier, C

    1987-11-07

    The diagnostic value of high sensitivity thyrotrophin assay (HS-TSH) and that of ultra-short determination of total thyroxine (T4) by fluorescence polarization (T4-TDX) were evaluated in 284 patients (29 hyperthyroid, 137 euthyroid without treatment and 118 under substitutive therapy). After comparing the clinician's first impression with the results of these laboratory tests the proportion of correctly classified untreated euthyroid patients was the same with T4-TDX as with HS-TSH (90%). In contrast, this proportion in hyperthyroid patients was about twice as low with HS-TSH (23%) as with T4-TDX (42%). While HS-TSH gives a better diagnostic evaluation, T4-TDX is an excellent indicator of the degree of hyperthyroidism. In patients under suppressive therapy HS-TSH and T4-TDX provide a better evaluation of therapeutic effectiveness.

  2. Immunoradiometric assay for quantification of prostate-specific antigen (PSA)

    Energy Technology Data Exchange (ETDEWEB)

    Ruiz, Miriam [Institute of Nuclear Sciences and Technology, Havana (Cuba). Dept.of Radiochemistry]. E-mail: mirian@fctn.isctn.edu.cu

    2002-07-01

    To develop an immunoradiometric assay (IRMA) for quantification of total PSA in serum, we carried out the evaluation of a panel of 12 monoclonal antibodies (MAbs). The mAbs (CB-PSA 1 to CB-PSA 10) were developed in the Center of Genetic Engineering and Biotechnology (CIGB) Havana, Cuba and mAbs (CMC-H9, CMC-A5) in the Center of Immunology and Biologics (CENIB), Camaguey, Cuba. The inhibition assays were carried out using as reference a commercial equimolar assay for total PSA. For the evaluation, the mAbs were labeled with {sup 125} I by the method of Chloramine T and the capture mAbs were used conjugated with biotin. For separation, it was used the avidin coupled to magnetic particles. To validate the assay we evaluated the sensibility, inter and intraassay precision and the correlation with reference assay in 65 samples of patient under suspicion of prostate cancer. The partner CB-PSA 4 / CB-PSA 9 (capture / tracer) showed the best results in the IRMA. The developed assay presented a detection limit of 0.025 mg/L, a good intra and interassay precision and a high correlation with the reference assay. (author)

  3. Quantitative Fissile Assay In Used Fuel Using LSDS System

    Directory of Open Access Journals (Sweden)

    Lee YongDeok

    2017-01-01

    Full Text Available A quantitative assay of isotopic fissile materials (U235, Pu239, Pu241 was done at Korea Atomic Energy Research Institute (KAERI, using lead slowing down spectrometer (LSDS. The optimum design of LSDS was performed based on economics, easy maintenance and assay effectiveness. LSDS system consists of spectrometer, neutron source, detection and control. LSDS system induces fissile fission and fast neutrons are collected at fission chamber. The detected signal has a direct relation to the mass of existing fissile isotopes. Many current commercial assay technologies have a limitation in direct application on isotopic fissile assay of spent fuel, except chemical analysis. In the designed system, the fissile assay model was setup and the correction factor for self-shield was obtained. The isotopic fissile content assay was performed by changing the content of Pu239. Based on the fuel rod, the isotopic content was consistent with ~2% uncertainty for Pu239. By applying the covering (neutron absorber, the effective shielding was obtained and the activation was calculated on the target. From the assay evaluation, LSDS technique is very powerful and direct to analyze the isotopic fissile content. LSDS is applicable for nuclear fuel cycle and spent fuel management for safety and economics. Additionally, an accurate fissile content will contribute to the international transparency and credibility on spent fuel.

  4. Traditional and Model Based Assay of Irregular Geometry Items

    Energy Technology Data Exchange (ETDEWEB)

    MOORE, FRANK S.; SALAYMEH, SALEEM

    2005-06-15

    The Analytical Development Section (ADS) of SRNL was requested to perform a waste disposal assay of two heater boxes which had been used in the HB Line dissolvers. They had been sent to SRNL for study to make recommendations on how to prevent future failure of the units when they were replaced. The study having been completed, the units needed to be characterized prior to sending to Solid Waste for disposal. An assay station consisting of a turntable, HPGe detector, CANBERRA Inspector, transmission source and a portable computer was set up to do the required assays. The assays indicate the presence of U-235, Pu-239 and Cs-137. No measurable amounts of U-235 or Pu-239 were found. Therefore the Minimum Detectable Activities for U-235 and Pu-239 were calculated. For Heater Box 1, 0.23 grams of U-235 and 0.24 grams of Pu-239. For Heater Box 2, the results were 0.21 grams of U-235 and 0.21 grams of Pu-239. This paper describes and documents the assays employed to determine the amount of U, Pu and Cs contents of the heater boxes. The paper provides results of SNM assays using traditional calibration of the system and on one based on modeling. It also provides the scientific community with data that will assist the user in determining the method of choice for assaying items with irregular geometries.

  5. Quantitative Fissile Assay In Used Fuel Using LSDS System

    Science.gov (United States)

    Lee, YongDeok; Jeon, Ju Young; Park, Chang-Je

    2017-09-01

    A quantitative assay of isotopic fissile materials (U235, Pu239, Pu241) was done at Korea Atomic Energy Research Institute (KAERI), using lead slowing down spectrometer (LSDS). The optimum design of LSDS was performed based on economics, easy maintenance and assay effectiveness. LSDS system consists of spectrometer, neutron source, detection and control. LSDS system induces fissile fission and fast neutrons are collected at fission chamber. The detected signal has a direct relation to the mass of existing fissile isotopes. Many current commercial assay technologies have a limitation in direct application on isotopic fissile assay of spent fuel, except chemical analysis. In the designed system, the fissile assay model was setup and the correction factor for self-shield was obtained. The isotopic fissile content assay was performed by changing the content of Pu239. Based on the fuel rod, the isotopic content was consistent with 2% uncertainty for Pu239. By applying the covering (neutron absorber), the effective shielding was obtained and the activation was calculated on the target. From the assay evaluation, LSDS technique is very powerful and direct to analyze the isotopic fissile content. LSDS is applicable for nuclear fuel cycle and spent fuel management for safety and economics. Additionally, an accurate fissile content will contribute to the international transparency and credibility on spent fuel.

  6. New low-viscosity overlay medium for viral plaque assays

    Directory of Open Access Journals (Sweden)

    Garten Wolfgang

    2006-08-01

    Full Text Available Abstract Background Plaque assays in cell culture monolayers under solid or semisolid overlay media are commonly used for quantification of viruses and antiviral substances. To overcome the pitfalls of known overlays, we tested suspensions of microcrystalline cellulose Avicel RC/CL™ as overlay media in the plaque and plaque-inhibition assay of influenza viruses. Results Significantly larger plaques were formed under Avicel-containing media, as compared to agar and methylcellulose (MC overlay media. The plaque size increased with decreasing Avicel concentration, but even very diluted Avicel overlays (0.3% ensured formation of localized plaques. Due to their low viscosity, Avicel overlays were easier to use than methylcellulose overlays, especially in the 96-well culture plates. Furthermore, Avicel overlay could be applied without prior removal of the virus inoculum thus facilitating the assay and reducing chances of cross-contamination. Using neuraminidase inhibitor oseltamivir carboxylate, we demonstrated applicability of the Avicel-based plaque reduction assay for testing of antiviral substances. Conclusion Plaque assay under Avicel-containing overlay media is easier, faster and more sensitive than assays under agar- and methylcellulose overlays. The assay can be readily performed in a 96-well plate format and seems particularly suitable for high-throughput virus titrations, serological studies and experiments on viral drug sensitivity. It may also facilitate work with highly pathogenic agents performed under hampered conditions of bio-safety labs.

  7. The application of protein microarray assays in psychoneuroimmunology.

    Science.gov (United States)

    Ayling, K; Bowden, T; Tighe, P; Todd, I; Dilnot, E M; Negm, O H; Fairclough, L; Vedhara, K

    2017-01-01

    Protein microarrays are miniaturized multiplex assays that exhibit many advantages over the commonly used enzyme-linked immunosorbent assay (ELISA). This article aims to introduce protein microarrays to readers of Brain, Behavior, and Immunity and demonstrate its utility and validity for use in psychoneuroimmunological research. As part of an ongoing investigation of psychological and behavioral influences on influenza vaccination responses, we optimized a novel protein microarray to quantify influenza-specific antibody levels in human sera. Reproducibility was assessed by calculating intra- and inter-assay coefficients of variance on serially diluted human IgG concentrations. A random selection of samples was analyzed by microarray and ELISA to establish validity of the assay. For IgG concentrations, intra-assay and inter-assay precision profiles demonstrated a mean coefficient of variance of 6.7% and 11.5% respectively. Significant correlations were observed between microarray and ELISA for all antigens, demonstrating the microarray is a valid alternative to ELISA. Protein microarrays are a highly robust, novel assay method that could be of significant benefit for researchers working in psychoneuroimmunology. They offer high throughput, fewer resources per analyte and can examine concurrent neuro-immune-endocrine mechanisms.

  8. Comparative endpoint sensitivity of in vitro estrogen agonist assays.

    Science.gov (United States)

    Dreier, David A; Connors, Kristin A; Brooks, Bryan W

    2015-07-01

    Environmental and human health implications of endocrine disrupting chemicals (EDCs), particularly xenoestrogens, have received extensive study. In vitro assays are increasingly employed as diagnostic tools to comparatively evaluate chemicals, whole effluent toxicity and surface water quality, and to identify causative EDCs during toxicity identification evaluations. Recently, the U.S. Environmental Protection Agency (USEPA) initiated ToxCast under the Tox21 program to generate novel bioactivity data through high throughput screening. This information is useful for prioritizing chemicals requiring additional hazard information, including endocrine active chemicals. Though multiple in vitro and in vivo techniques have been developed to assess estrogen agonist activity, the relative endpoint sensitivity of these approaches and agreement of their conclusions remain unclear during environmental diagnostic applications. Probabilistic hazard assessment (PHA) approaches, including chemical toxicity distributions (CTD), are useful for understanding the relative sensitivity of endpoints associated with in vitro and in vivo toxicity assays by predicting the likelihood of chemicals eliciting undesirable outcomes at or above environmentally relevant concentrations. In the present study, PHAs were employed to examine the comparative endpoint sensitivity of 16 in vitro assays for estrogen agonist activity using a diverse group of compounds from the USEPA ToxCast dataset. Reporter gene assays were generally observed to possess greater endpoint sensitivity than other assay types, and the Tox21 ERa LUC BG1 Agonist assay was identified as the most sensitive in vitro endpoint for detecting an estrogenic response. When the sensitivity of this most sensitive ToxCast in vitro endpoint was compared to the human MCF-7 cell proliferation assay, a common in vitro model for biomedical and environmental monitoring applications, the ERa LUC BG1 assay was several orders of magnitude less

  9. The EHEC-crisis - Impact and lessons learned - Sustainable cross-border crisis control and communication

    NARCIS (Netherlands)

    Hamer, M.; Terlau, W.; Terlau, W.; Roest, van der J.; Petersen, B.

    2016-01-01

    Food-borne disease is an ever-present threat and is often associated with the consumption of fresh food such as horticultural products. Apart from chemicals, heavy metals, fungi and viruses, bacteria are the most common cause for food poisoning. A serious outbreak in Germany and neighbouring

  10. Facile and Sensitive Epifluorescent Silica Nanoparticles for the Rapid Screening of EHEC

    Directory of Open Access Journals (Sweden)

    Pravate Tuitemwong

    2013-01-01

    Full Text Available This study was to develop antibodies conjugated fluorescent dye-doped silica nanoparticles (FDS-NPs aiming to increase signals for the rapid detection of Escherichia coli O157:H7 with glass slide method. The FDS-NPs were produced with microemulsion/sol-gel techniques resulting in spherical in shape with 47 ± 6 nm in diameter. The particles showed high intensity and stable orange color Rubpy luminescent dye. The XRD spectrum showed a broad diffraction peak in the range of – (centered at indicating an amorphous structure. Surface modifications for bioconjugation with affinity chromatography purified (IgGs antibodies were successful. The properties were evident from FTIR spectra at 1631.7 . Results indicated that nanoparticles could attach onto cells of E. coli O157:H7 coated on a glass slide, and give distinctively bright color under epifluorescence microscope (400x. It was shown that FDS-NPs could detect a very low amount of cells of E. coli O157:H7 (16 CFU in 10 ml in 60 min. The phosphate buffered saline (PBS with ionic strength of 1.70 gave zeta potential of good particle dispersion (−40 mV. This work demonstrated that highly sensitive bioconjugated E. coli O157:H7 FDS-NPs were successfully developed with a potential to be used for the rapid detection of E. coli O157:H7 in foods.

  11. The EHEC-crisis - Impact and lessons learned - Sustainable cross-border crisis control and communication

    NARCIS (Netherlands)

    Hamer, M.; Terlau, W.; Terlau, W.; Roest, van der J.; Petersen, B.

    2016-01-01

    Food-borne disease is an ever-present threat and is often associated with the consumption of fresh food such as horticultural products. Apart from chemicals, heavy metals, fungi and viruses, bacteria are the most common cause for food poisoning. A serious outbreak in Germany and neighbouring coun

  12. Droplet microfluidics for high-throughput biological assays.

    Science.gov (United States)

    Guo, Mira T; Rotem, Assaf; Heyman, John A; Weitz, David A

    2012-06-21

    Droplet microfluidics offers significant advantages for performing high-throughput screens and sensitive assays. Droplets allow sample volumes to be significantly reduced, leading to concomitant reductions in cost. Manipulation and measurement at kilohertz speeds enable up to 10(8) samples to be screened in one day. Compartmentalization in droplets increases assay sensitivity by increasing the effective concentration of rare species and decreasing the time required to reach detection thresholds. Droplet microfluidics combines these powerful features to enable currently inaccessible high-throughput screening applications, including single-cell and single-molecule assays.

  13. A rapid and efficient assay for extracting DNA from fungi

    Science.gov (United States)

    Griffin, Dale W.; Kellogg, C.A.; Peak, K.K.; Shinn, E.A.

    2002-01-01

    Aims: A method for the rapid extraction of fungal DNA from small quantities of tissue in a batch-processing format was investigated. Methods and Results: Tissue (DNA for PCR/ sequencing applications. Conclusions: The method allowed batch DNA extraction from multiple fungal isolates using a simple yet rapid and reliable assay. Significance and Impact of the Study: Use of this assay will allow researchers to obtain DNA from fungi quickly for use in molecular assays that previously required specialized instrumentation, was time-consuming or was not conducive to batch processing.

  14. A non-isotopic assay for histone deacetylase activity.

    Science.gov (United States)

    Hoffmann, K; Brosch, G; Loidl, P; Jung, M

    1999-05-01

    Inhibitors of histone deacetylase (HD) bear great potential as new drugs due to their ability to modulate transcription and to induce apoptosis or differentiation in cancer cells. To study the activity of HD and the effect of potential inhibitors in vitro so far only radio-active assays have existed. For the search of new inhibitors and for the use in HD identification and purification we established a simple, non-radioactive assay that allows screening of large numbers of compounds. The assay is based on an aminocoumarin derivative of an Omega-acetylated lysine as enzyme substrate.

  15. A Sensitive Chemotaxis Assay Using a Novel Microfluidic Device

    Directory of Open Access Journals (Sweden)

    Chen Zhang

    2013-01-01

    Full Text Available Existing chemotaxis assays do not generate stable chemotactic gradients and thus—over time—functionally measure only nonspecific random motion (chemokinesis. In comparison, microfluidic technology has the capacity to generate a tightly controlled microenvironment that can be stably maintained for extended periods of time and is, therefore, amenable to adaptation for assaying chemotaxis. We describe here a novel microfluidic device for sensitive assay of cellular migration and show its application for evaluating the chemotaxis of smooth muscle cells in a chemokine gradient.

  16. Advances in Assays and Analytical Approaches for Botulinum Toxin Detection

    Energy Technology Data Exchange (ETDEWEB)

    Grate, Jay W.; Ozanich, Richard M.; Warner, Marvin G.; Bruckner-Lea, Cindy J.; Marks, James D.

    2010-08-04

    Methods to detect botulinum toxin, the most poisonous substance known, are reviewed. Current assays are being developed with two main objectives in mind: 1) to obtain sufficiently low detection limits to replace the mouse bioassay with an in vitro assay, and 2) to develop rapid assays for screening purposes that are as sensitive as possible while requiring an hour or less to process the sample an obtain the result. This review emphasizes the diverse analytical approaches and devices that have been developed over the last decade, while also briefly reviewing representative older immunoassays to provide background and context.

  17. Ethanal assay, using an enzymo-conductimetric method

    Energy Technology Data Exchange (ETDEWEB)

    Saad, I.; Wallach, J.M. (I.C.B.M.C., Villeurbanne (France))

    1992-01-01

    An enzymo-conductimetric method for the assay of ethanol is described. It is based on ethanol oxidation is presence of yeast aldehyde dehydrogenase. Experimental conditions compatible with conductimetry were optimized and kinetic parameters of the enzymatic reaction determined. Due to the low K{sub M} value of ethanol for the enzyme, an end-point assay was proposed. A linear relationship was demonstrated between experimental conductance changes and ethanol concentration up to 25 {mu}M. Validation of the assay was made on wines by comparison with a spectrophotometric method. Both methods gave similar results, but conductimetry avoids sample treatment if solutions are colored or turbid.

  18. Application of radioreceptor assay of benzodiazepines for toxicology

    Energy Technology Data Exchange (ETDEWEB)

    Aaltonen, L.; Scheinin, M. (Department of Pharmacology and the Department of Internal Medicine, University of Turku, Finland)

    1982-01-01

    A radioreceptor assay (RRA) for determining benzodiazepines (BZ) has been developed and applied to toxicological analysis of serum samples from 21 patients with acute BZ overdosage. The method was sensitive (e.g., lorazepam 17 nM, diazepam 41 nM), and specific for pharmacologically active BZ derivatives. The reproducibility of the results was good (intra-assay variation < 8%, inter-assay variation < 10%). Concentrations measured by the RRA showed a good correlation with those obtained by gas-liquid chromatographic analysis of the same samples. The quantitative results represent the sum of one or several parent substances and all biologically active metabolites, in proportion to their receptor binding affinities.

  19. De David Belasco à Giacomo Puccini : La Fanciulla del West, premier opéra « américain » From David Belasco to Giacomo Puccini:La Fanciulla del West, first “American” opera

    Directory of Open Access Journals (Sweden)

    Walter Zidaric

    2009-10-01

    Full Text Available La Fanciulla del West is a “horse opera” resulting from the second collaboration between David Belasco and Giacomo Puccini. This work, specially composed for the American stage, which had its world premiere at New York’s Metropolitan Opera in December 1910, paved the way for American composers who increasingly focused on American History in their quest for a “national” opera. For the first time, Puccini resorted to genuine Indian music, a Zuni melody, which he re-elaborated for dramatic purposes and associated with the nostalgic mood of uprooted people, the miners who left their home country for America. The Other – the native American viewed from the vantage point of the white, as well as the foreigner – no longer represented a threat, as had been the case in the operatic repertoire before, but became part and parcel of a multicultural society, like the Italian immigrant to whom Puccini also paid tribute. The Zuni melody, which opens and closes the opera, contributes to giving a genuine twist to a score which is the first one to tackle the foundations of the American nation.

  20. Application of Adaptation Theory in Practice of Interpreting Taking the Interpreting Reports of Premier WEN in 2010 .as an Example%语言顺应论在口译实践中的应用——以2010年温家宝总理答记者问现场口译为例

    Institute of Scientific and Technical Information of China (English)

    蒋瑛

    2011-01-01

    Adaptation theory was proposed by Versehueren in 2000, the main idea of which is that language has three properties : variability, negotiability and adaptability. Based on the adaptation theory, how the target language adapts to the context, language structure and the dynamic process of interpreting were explained to demonstrate the important guiding function through studying the interpreting reports of Premier WEN in 2010.%语言顺应论是Verschueren于2000年提出,之后被广泛应用于语言学界,其主要内容包括变异性、商讨性和顺应性。以语言顺应论为理论支撑,以2010年温总理答记者问现场口译稿为语料,分析口译实践过程中语言顺应论如何解释译语顺应语境、语言结构和口译的动态过程,说明语言顺应论在口译实践过程中的重要指导作用。

  1. Comparative evaluation under routine conditions of the nitrate reduction assay, the proportion assay and the MGIT 960 assay for drug susceptibility testing of clinical isolates of Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Leila de Souza Fonseca

    2012-02-01

    Full Text Available The performance of the nitrate reductase assay (NRA was compared with the proportion method (PM on Lowenstein-Jensen medium and the BACTEC MGIT960 assay under routine conditions using 160 clinical isolates of Mycobacterium tuberculosis with a high proportion of resistant strains. The mean time to obtain results was 8.8 days and the overall agreements between NRA and PM and NRA and M960 were 95% and 94%, respectively. NRA was easy to perform and represents a useful tool for the rapid screening of drug-resistant M. tuberculosis strains in low-resource countries.

  2. Development of a rapid agglutination latex test for diagnosis of enteropathogenic and enterohemorrhagic Escherichia coli infection in developing world: defining the biomarker, antibody and method.

    Directory of Open Access Journals (Sweden)

    Letícia B Rocha

    2014-09-01

    Full Text Available Enteropathogenic and enterohemorrhagic Escherichia coli (EPEC/EHEC are human intestinal pathogens responsible for diarrhea in both developing and industrialized countries. In research laboratories, EPEC and EHEC are defined on the basis of their pathogenic features; nevertheless, their identification in routine laboratories is expensive and laborious. Therefore, the aim of the present work was to develop a rapid and simple assay for EPEC/EHEC detection. Accordingly, the EPEC/EHEC-secreted proteins EspA and EspB were chosen as target antigens.First, we investigated the ideal conditions for EspA/EspB production/secretion by ELISA in a collection of EPEC/EHEC strains after cultivating bacterial isolates in Dulbecco's modified Eagle's medium (DMEM or DMEM containing 1% tryptone or HEp-2 cells-preconditioned DMEM, employing either anti-EspA/anti-EspB polyclonal or monoclonal antibodies developed and characterized herein. Subsequently, a rapid agglutination latex test (RALT was developed and tested with the same collection of bacterial isolates.EspB was defined as a biomarker and its corresponding monoclonal antibody as the tool for EPEC/EHEC diagnosis; the production of EspB was better in DMEM medium. RALT assay has the sensitivity and specificity required for high-impact diagnosis of neglected diseases in the developing world.RALT assay described herein can be considered an alternative assay for diarrhea diagnosis in low-income countries since it achieved 97% sensitivity, 98% specificity and 97% efficiency.

  3. Microfluidic Assay to Quantify the Adhesion of Marine Bacteria

    National Research Council Canada - National Science Library

    Arpa-Sancet, M P; Christophis, C; Rosenhahn, A

    2012-01-01

    .... To determine the attachment strength of bacteria to coatings, a microfluidic adhesion assay has been developed which allows probing at which critical wall shear stress bacteria are removed from the surface...

  4. Quantitative Assays for RAS Pathway Proteins and Phosphorylation States

    Science.gov (United States)

    The NCI CPTAC program is applying its expertise in quantitative proteomics to develop assays for RAS pathway proteins. Targets include key phosphopeptides that should increase our understanding of how the RAS pathway is regulated.

  5. Radiometric microbiologic assay for the biologically active forms of niacin

    Energy Technology Data Exchange (ETDEWEB)

    Kertcher, J.A.; Guilarte, T.R.; Chen, M.F.; Rider, A.A.; McIntyre, P.A.

    1979-05-01

    A radiometric microbiologic assay has been developed for the determination of niacin in biologic fluids. Lactobacillus plantarum produced /sup 14/CO/sub 2/ from L-(U-/sup 14/C) malic acid in quantities proportional to the amount of niacin present. The assay is specific for the biologically active forms of niacin in humans. Thirty normal hemolysates were analyzed and the values ranged from 13.0 to 17.8 ..mu..g niacin/ml RBC (mean = 15.27 +- 1.33 s.d.). Good recovery and reproducibility studies were obtained with this assay. On thirty blood samples, correlation was excellent between the radiometric and the conventional turbidimetric assays.

  6. Effect of Spacer and the Enzyme-Linked Immunosorbent Assay

    Directory of Open Access Journals (Sweden)

    Manisha Sathe

    2016-09-01

    Full Text Available The effect of spacers and the enzyme-linked immunosorbent assay (ELISA formats on the functional parameters of assays such as lower detection limit, inhibitory concentration at 50 per cent (IC50, and specificity were studied. Enzyme conjugates having hydrophobic and hydrophilic spacers were prepared using O-isopropyl methylphosphonic acid (IMPA and horseradish peroxidase (HRP as an enzyme label. Comparison was made with reference to enzyme conjugate without any spacer. The present investigation revealed that the presence of a hydrophilic spacer in the enzyme conjugate significantly improves the sensitivity of assays. An enhanced IC50 value achieved was 0.01 μg mL−1 for free antigen detection by direct immunoassay using hydrophilic spacers and precoating of ELISA plates by secondary antibody. The use of a hydrophilic spacer might have helped in projecting the hapten in the aqueous phase, leading to enhanced antibody binding signal and improved sensitivity of the assay.

  7. Sperm penetration assay and its correlation with semen analysis parameters

    Directory of Open Access Journals (Sweden)

    Laxmi Kant Pandey

    2015-11-01

    Full Text Available Background: Aim of current study was to determine whether the Sperm Penetration Assay (SPA can be used as a test to discriminate the infertile male from fertile one. We have also correlated the SPA with semen analysis. Methods: Sperm characteristics namely Semen analysis and the sperm penetration assay were tested in 44 infertile and 10 fertile men. Sperm penetration assay was determined by using zona free hamster eggs. Results: With decreasing spermatozoa concentration in the semen there was significant decrease in percentage penetration of zona free Hamster eggs (p0.05. Conclusions: The Sperm penetration assay could discriminate the infertile group from fertile group significantly (p<0.001. The test appeared to be highly reproducible and probably identifies a truly infertile male. [Int J Res Med Sci 2015; 3(11.000: 3197-3201

  8. Alkaline Comet Assay for Assessing DNA Damage in Individual Cells.

    Science.gov (United States)

    Pu, Xinzhu; Wang, Zemin; Klaunig, James E

    2015-08-06

    Single-cell gel electrophoresis, commonly called a comet assay, is a simple and sensitive method for assessing DNA damage at the single-cell level. It is an important technique in genetic toxicological studies. The comet assay performed under alkaline conditions (pH >13) is considered the optimal version for identifying agents with genotoxic activity. The alkaline comet assay is capable of detecting DNA double-strand breaks, single-strand breaks, alkali-labile sites, DNA-DNA/DNA-protein cross-linking, and incomplete excision repair sites. The inclusion of digestion of lesion-specific DNA repair enzymes in the procedure allows the detection of various DNA base alterations, such as oxidative base damage. This unit describes alkaline comet assay procedures for assessing DNA strand breaks and oxidative base alterations. These methods can be applied in a variety of cells from in vitro and in vivo experiments, as well as human studies.

  9. Transient expression assays in grapevine: a step towards genetic improvement

    National Research Council Canada - National Science Library

    Jelly, Noémie S; Valat, Laure; Walter, Bernard; Maillot, Pascale

    2014-01-01

    In the past few years, the usefulness of transient expression assays has continuously increased for the characterization of unknown gene function and metabolic pathways. In grapevine ( Vitis vinifera L...

  10. Optimising automation of a manual enzyme-linked immunosorbent assay

    National Research Council Canada - National Science Library

    Corena de Beer; Monika Esser; Wolfgang Preiser

    2011-01-01

    .... Compared to conventional manual assays, automated ELISA systems offer more accurate and reproducible results, faster turnaround times and cost effectiveness due to the use of multianalyte reagents.Design: The VaccZyme...

  11. PubChem BioAssay: 2017 update

    Science.gov (United States)

    Wang, Yanli; Bryant, Stephen H.; Cheng, Tiejun; Wang, Jiyao; Gindulyte, Asta; Shoemaker, Benjamin A.; Thiessen, Paul A.; He, Siqian; Zhang, Jian

    2017-01-01

    PubChem's BioAssay database (https://pubchem.ncbi.nlm.nih.gov) has served as a public repository for small-molecule and RNAi screening data since 2004 providing open access of its data content to the community. PubChem accepts data submission from worldwide researchers at academia, industry and government agencies. PubChem also collaborates with other chemical biology database stakeholders with data exchange. With over a decade's development effort, it becomes an important information resource supporting drug discovery and chemical biology research. To facilitate data discovery, PubChem is integrated with all other databases at NCBI. In this work, we provide an update for the PubChem BioAssay database describing several recent development including added sources of research data, redesigned BioAssay record page, new BioAssay classification browser and new features in the Upload system facilitating data sharing. PMID:27899599

  12. Usefulness of human coagulation and fibrinolysis assays in domestic pigs

    DEFF Research Database (Denmark)

    Münster, Anna-Marie Bloch; Olsen, Aage Kristian; Bladbjerg, Else-Marie

    2002-01-01

    Pigs are often used as animal models in research on blood coagulation and fibrinolysis. The usefulness of the assays applied within this field, and the knowledge of reference intervals are therefore essential and of utmost importance. In the study reported here, we investigated the applicability...... time, tissue factor, tissue factor pathway inhibitor, factor VII, protein C, protein S, prothrombin fragment 1+2, antithrombin, thrombin-antithrombin complexes, fibrinogen, soluble fibrin, urokinase-type plasminogen activator, plasmin inhibitor, plasminogen activator inhibitor 1, and D-dimer. We found...... that 11 of 12 functional assays, but only 3 of 10 immunoassays, were applicable to porcine plasma, and we determined the normal range of these variables. We conclude that human functional assays are useful in porcine plasma, whereas only a few immunologic assays can be used. However, precautions must...

  13. Development of fluorescent methods for DNA methyltransferase assay

    Science.gov (United States)

    Li, Yueying; Zou, Xiaoran; Ma, Fei; Tang, Bo; Zhang, Chun-yang

    2017-03-01

    DNA methylation modified by DNA methyltransferase (MTase) plays an important role in regulating gene transcription, cell growth and proliferation. The aberrant DNA MTase activity may lead to a variety of human diseases including cancers. Therefore, accurate and sensitive detection of DNA MTase activity is crucial to biomedical research, clinical diagnostics and therapy. However, conventional DNA MTase assays often suffer from labor-intensive operations and time-consuming procedures. Alternatively, fluorescent methods have significant advantages of simplicity and high sensitivity, and have been widely applied for DNA MTase assay. In this review, we summarize the recent advances in the development of fluorescent methods for DNA MTase assay. These emerging methods include amplification-free and the amplification-assisted assays. Moreover, we discuss the challenges and future directions of this area.

  14. Awareness of antimullerian hormone assay and its relevance in in ...

    African Journals Online (AJOL)

    Awareness of antimullerian hormone assay and its relevance in in-vitro fertilization ... especially in areas of current methods for successful IVF treatment. Key words: Antimullerian hormone, In Vitro Fertilization, Infertility, Laboratory scientist ...

  15. Click Chemistry-Mediated Nanosensors for Biochemical Assays

    Science.gov (United States)

    Chen, Yiping; Xianyu, Yunlei; Wu, Jing; Yin, Binfeng; Jiang, Xingyu

    2016-01-01

    Click chemistry combined with functional nanoparticles have drawn increasing attention in biochemical assays because they are promising in developing biosensors with effective signal transformation/amplification and straightforward signal readout for clinical diagnostic assays. In this review, we focus on the latest advances of biochemical assays based on Cu (I)-catalyzed 1, 3-dipolar cycloaddition of azides and alkynes (CuAAC)-mediated nanosensors, as well as the functionalization of nanoprobes based on click chemistry. Nanoprobes including gold nanoparticles, quantum dots, magnetic nanoparticles and carbon nanomaterials are covered. We discuss the advantages of click chemistry-mediated nanosensors for biochemical assays, and give perspectives on the development of click chemistry-mediated approaches for clinical diagnosis and other biomedical applications. PMID:27217831

  16. Piceance Basin Oil Shale Data: Assays, Boreholes and Formation Tops

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — This database contains Oil Shale Assays, Borehole Locations and Formation Tops that were used in support of the 2009 Oil Shale Assessment (Survey Fact Sheet...

  17. Mouse Assay for Determination of Arsenic Bioavailability in Contaminated Soils

    Science.gov (United States)

    Background: Accurate assessment of human exposure estimates from arsenic-contaminated soils depends upon estimating arsenic (As) soil bioavailability. Development of bioavailability assays provides data needed for human health risk assessments and supports development and valida...

  18. Cell-based Assays to Identify Inhibitors of Viral Disease

    Science.gov (United States)

    Green, Neil; Ott, Robert D.; Isaacs, Richard J.; Fang, Hong

    2009-01-01

    Background Antagonizing the production of infectious virus inside cells requires drugs that can cross the cell membrane without harming host cells. Objective It is therefore advantageous to establish intracellular potency of anti-viral drug candidates early in the drug-discovery pipeline. Methods To this end, cell-based assays are being developed and employed in high-throughput drug screening, ranging from assays that monitor replication of intact viruses to those that monitor activity of specific viral proteins. While numerous cell-based assays have been developed and investigated, rapid counter screens are also needed to define the specific viral targets of identified inhibitors and to eliminate nonspecific screening hits. Results/Conclusions Here, we describe the types of cell-based assays being used in antiviral drug screens and evaluate the equally important counter screens that are being employed to reach the full potential of cell-based high-throughput screening. PMID:19750206

  19. Click Chemistry-Mediated Nanosensors for Biochemical Assays.

    Science.gov (United States)

    Chen, Yiping; Xianyu, Yunlei; Wu, Jing; Yin, Binfeng; Jiang, Xingyu

    2016-01-01

    Click chemistry combined with functional nanoparticles have drawn increasing attention in biochemical assays because they are promising in developing biosensors with effective signal transformation/amplification and straightforward signal readout for clinical diagnostic assays. In this review, we focus on the latest advances of biochemical assays based on Cu (I)-catalyzed 1, 3-dipolar cycloaddition of azides and alkynes (CuAAC)-mediated nanosensors, as well as the functionalization of nanoprobes based on click chemistry. Nanoprobes including gold nanoparticles, quantum dots, magnetic nanoparticles and carbon nanomaterials are covered. We discuss the advantages of click chemistry-mediated nanosensors for biochemical assays, and give perspectives on the development of click chemistry-mediated approaches for clinical diagnosis and other biomedical applications.

  20. Influences of acidic conditions on formazan assay: a cautionary note.

    Science.gov (United States)

    Johno, Hisashi; Takahashi, Shuhei; Kitamura, Masanori

    2010-11-01

    Formazan assay has been used for several decades to evaluate metabolic activity of eukaryotic and prokaryotic cells. In particular, it has been often applied for quantitative assessment of viable cells under acidic circumstances caused by, e.g., ischemia and hypoxia. However, little attention has been paid to the influence of acidic pH on formazan assays. We found that acidic culture conditions significantly affect outcomes of the assays. Absorbance of tetrazolium-formazan decreased in a pH-dependent manner without affecting cell viability. This nonspecific effect was ascribed to influences of acidic pH on the production of formazan. Replacement of culture media to fresh medium at physiologic pH partially overcame this problem. The influence of acidic culture conditions should be carefully considered when formazan assays are used for the assessment of viable cells under various experimental situations.

  1. A novel behavioral assay for measuring cold sensation in mice.

    Directory of Open Access Journals (Sweden)

    Daniel S Brenner

    Full Text Available Behavioral models of cold responses are important tools for exploring the molecular mechanisms of cold sensation. To complement the currently cold behavioral assays and allow further studies of these mechanisms, we have developed a new technique to measure the cold response threshold, the cold plantar assay. In this assay, animals are acclimated on a glass plate and a cold stimulus is applied to the hindpaw through the glass using a pellet of compressed dry ice. The latency to withdrawal from the cooled glass is used as a measure of the cold response threshold of the rodents, and the dry ice pellet provides a ramping cold stimulus on the glass that allows the correlation of withdrawal latency values to rough estimates of the cold response threshold temperature. The assay is highly sensitive to manipulations including morphine-induced analgesia, Complete Freund's Adjuvant-induced inflammatory allodynia, and Spinal Nerve Ligation-induced neuropathic allodynia.

  2. Lipid-binding analysis using a fat blot assay.

    NARCIS (Netherlands)

    Munnik, T.; Wierzchowiecka, M.

    2013-01-01

    Protein-lipid interactions play an important role in lipid metabolism, membrane trafficking and cell -signaling by regulating protein localization, activation, and function. The Fat Blot assay is a relatively simple and inexpensive method to examine these interactions using nitrocellulose

  3. Multiplexed Dosing Assays by Digitally Definable Hydrogel Volumes

    DEFF Research Database (Denmark)

    Faralli, Adele; Melander, Fredrik; Larsen, Esben Kjær Unmack

    2016-01-01

    Stable and low-cost multiplexed drug sensitivity assays using small volumes of cells or tissue are in demand for personalized medicine, including patientspecific combination chemotherapy. Spatially defined projected light photopolymerization of hydrogels with embedded active compounds is introduc...

  4. A Cell-Based Assay to Assess Hemichannel Function

    Science.gov (United States)

    Krishnan, Srinivasan; Fiori, Mariana C.; Cuello, Luis G.; Altenberg, Guillermo A.

    2017-01-01

    Activation of connexin hemichannels is involved in the pathophysiology of disorders that include deafness, stroke, and cardiac infarct. This aspect makes hemichannels an attractive therapeutic target. Unfortunately, most available inhibitors are not selective or isoform specific, which hampers their translational application. The absence of a battery of useful inhibitors is due in part to the absence of simple screening assays for the discovery of hemichannel-active drugs. Here, we present an assay that we have recently developed to assess hemichannel function. The assay is based on the expression of functional human connexins in a genetically modified bacterial strain deficient in K+ uptake. These modified cells do not grow in low-K+ medium, but functional expression of connexin hemichannels allows K+ uptake and growth. This cell-growth-based assay is simple, robust, and easily scalable to high-throughput multi-well platforms.

  5. Application of neutron multiplicity counting to waste assay

    Energy Technology Data Exchange (ETDEWEB)

    Pickrell, M.M.; Ensslin, N. [Los Alamos National Lab., NM (United States); Sharpe, T.J. [North Carolina State Univ., Raleigh, NC (United States)

    1997-11-01

    This paper describes the use of a new figure of merit code that calculates both bias and precision for coincidence and multiplicity counting, and determines the optimum regions for each in waste assay applications. A {open_quotes}tunable multiplicity{close_quotes} approach is developed that uses a combination of coincidence and multiplicity counting to minimize the total assay error. An example is shown where multiplicity analysis is used to solve for mass, alpha, and multiplication and tunable multiplicity is shown to work well. The approach provides a method for selecting coincidence, multiplicity, or tunable multiplicity counting to give the best assay with the lowest total error over a broad spectrum of assay conditions. 9 refs., 6 figs.

  6. Mouse Assay for Determination of Arsenic Bioavailability in Contaminated Soils

    Science.gov (United States)

    Background: Accurate assessment of human exposure estimates from arsenic-contaminated soils depends upon estimating arsenic (As) soil bioavailability. Development of bioavailability assays provides data needed for human health risk assessments and supports development and valida...

  7. The comet assay – from toy to tool

    Directory of Open Access Journals (Sweden)

    Guenter Speit

    2015-04-01

    Full Text Available The comet assay is nowadays the most common method for measuring DNA damage and repair in single cells. It is based on the microelectrophoretic study published by Ostling and Johanson (1984 and was developed by Singh and coworkers (1988 to a versatile technique for quantitation of low levels of DNA damage in individual cells. This alkaline version still is the basis for the triumphant success of the comet assay in basic research into mechanisms of DNA damage and DNA repair, genotoxicity testing, ecotoxicology and human biomonitoring. Important technical improvements (e.g., the use of precoated slides, introduction of image analysis, high throughput methods, automated scoring systems made the assay more robust and more efficient. Modifications of the standard protocol provide more specific information on the type and biological significance of the damage studied. The introduction of lesion-specific endonucleases allowed the characterization of oxidative base damage, alkylation damage and UV-induced pyrimidine dimers. The combination with fluorescence in situ hybridization (FISH made it possible to identify DNA of particular chromosome regions and measure effects of damage and repair in particular genes. The comet assay is being increasingly used in genotoxicity testing. In particular, the in vivo comet assay has become a component of some genotoxicity test strategies and generally accepted test protocols have evolved over the years. A large international collaborative trial sponsored by the Japanese Center for the Validation of Alternative Methods (JaCVAM was recently completed and an OECD test guideline was approved. The comet assay is widely used in human biomonitoring to measure DNA damage as a marker of exposure to genotoxic agents or to investigate genoprotective effects. However, there are still problems in comparing results from different laboratories and there is need for reducing inter-laboratory variation and identification of standard

  8. Spectrophotometric Assays of Major Compounds Extracted from Algae.

    Science.gov (United States)

    Connan, Solène

    2015-01-01

    This chapter describes spectrophotometric assays of major compounds extracted from microalgae and macroalgae, i.e., proteins, carbohydrates, pigments (chlorophylls, carotenoids, and phycobiliproteins) and phenolic compounds. In contrast to other specific analytical techniques, such as high pressure liquid chromatography (HPLC) or mass spectrometry (MS), commonly applied to purified extracts to reveal more detailed composition and structure of algal compound families, these assays serve as a first assessment of the global contents of extracts.

  9. Bioluminescence-Sensing Assay for Microbial Growth Recognition

    OpenAIRE

    Heba Ramadan Eed; Nora S. Abdel-Kader; Mahmoud Helmy El Tahan; Tianhong Dai; Rehab Amin

    2016-01-01

    The conventional methods for microbial viability quantification require cultivation and are laborious. There is consequently a widespread need for cultivation-free methods. The adenosine triphosphate (ATP) bioluminescence-sensing assay is considered an extremely effective biosensor; hence ATP is the energy currency of all living microbes and can be used as a rapid indicator of microbial viability. We developed an ATP bioluminescence-sensing assay to detect microbial viability. A biolumine...

  10. FLIPR assays of intracellular calcium in GPCR drug discovery

    DEFF Research Database (Denmark)

    Hansen, Kasper Bø; Bräuner-Osborne, Hans

    2009-01-01

    Fluorescent dyes sensitive to changes in intracellular calcium have become increasingly popular in G protein-coupled receptor (GPCR) drug discovery for several reasons. First of all, the assays using the dyes are easy to perform and are of low cost compared to other assays. Second, most non-Galph...... making them obtainable even for academic groups. Here, we present a protocol for measuring changes in intracellular calcium levels in living mammalian cells based on the fluorescent calcium binding dye, fluo-4....

  11. Monkey Feeding Assay for Testing Emetic Activity of Staphylococcal Enterotoxin.

    Science.gov (United States)

    Seo, Keun Seok

    2016-01-01

    Staphylococcal enterotoxins (SEs) are unique bacterial toxins that cause gastrointestinal toxicity as well as superantigenic activity. Since systemic administration of SEs induces superantigenic activity leading to toxic shock syndrome that may mimic enterotoxic activity of SEs such as vomiting and diarrhea, oral administration of SEs in the monkey feeding assay is considered as a standard method to evaluate emetic activity of SEs. This chapter summarizes and discusses practical considerations of the monkey feeding assay used in studies characterizing classical and newly identified SEs.

  12. Analytical applications of MIPs in diagnostic assays: future perspectives.

    Science.gov (United States)

    Bedwell, Thomas S; Whitcombe, Michael J

    2016-03-01

    Many efforts have been made to produce artificial materials with biomimetic properties for applications in binding assays. Among these efforts, the technique of molecular imprinting has received much attention because of the high selectivity obtainable for molecules of interest, robustness of the produced polymers, simple and short synthesis, and excellent cost efficiency. In this review, progress in the field of molecularly imprinted sorbent assays is discussed-with a focus on work conducted from 2005 to date.

  13. Fluorescence Assay for Evaluating Microbicidal Activity of Hand Antiseptics

    OpenAIRE

    Lopez-Gigosos, Rosa M.; Mariscal, Alberto; Mariscal-Lopez, Eloisa; Gutierrez-Bedmar, Mario; Fernandez, Joaquin

    2015-01-01

    We developed a fluorescent β-d-glucuronidase activity (BGA)-based assay for detecting and quantifying Escherichia coli in samples to assess the biocide efficacy of hand antiseptics. The fluorescence level is proportional to the number of viable E. coli organisms present. We compared our assay results to those of the E. coli plate count method specified by the European standard for testing hygienic hand rub disinfectant products (EN1500). The plate count method requires excessive handling and ...

  14. Antibiotic microbial assay using kinetic-reading microplate system

    OpenAIRE

    Felipe Rebello Lourenço; Terezinha de Jesus Andreoli Pinto

    2011-01-01

    The aim of this study was to determine the optimal experimental conditions to develop a methodology for microbiological assay of apramycin employing microplate and kinetic reading mode, and to validate the developed method, through evaluation of parameters of selectivity, linearity, linear range, limits of detection and quantification, accuracy and precision. The turbidimetric assay principle is simple: the test solution is added to a suspension of test microorganism in culture media, the mix...

  15. Multiplex PCR Assay for Identification of Human Diarrheagenic Escherichia coli

    OpenAIRE

    Toma, Claudia; Lu, Yan; Higa, Naomi; Nakasone, Noboru; Isabel CHINEN; Baschkier, Ariela; Rivas, Marta; Iwanaga, Masaaki

    2003-01-01

    A multiplex PCR assay for the identification of human diarrheagenic Escherichia coli was developed. The targets selected for each category were eae for enteropathogenic E. coli, stx for Shiga toxin-producing E. coli, elt and est for enterotoxigenic E. coli, ipaH for enteroinvasive E. coli, and aggR for enteroaggregative E. coli. This assay allowed the categorization of a diarrheagenic E. coli strain in a single reaction tube.

  16. Optimising automation of a manual enzyme-linked immunosorbent assay

    OpenAIRE

    Corena de Beer; Monika Esser; Wolfgang Preiser

    2011-01-01

    Objective: Enzyme-linked immunosorbent assays (ELISAs) are widely used to quantify immunoglobulin levels induced by infection or vaccination. Compared to conventional manual assays, automated ELISA systems offer more accurate and reproducible results, faster turnaround times and cost effectiveness due to the use of multianalyte reagents.Design: The VaccZyme™ Human Anti-Haemophilus influenzae type B (Hib) kit (MK016) from The Binding Site Company was optimised to be used on an automated BioRad...

  17. Diced electrophoresis gel assay for screening enzymes with specified activities.

    Science.gov (United States)

    Komatsu, Toru; Hanaoka, Kenjiro; Adibekian, Alexander; Yoshioka, Kentaro; Terai, Takuya; Ueno, Tasuku; Kawaguchi, Mitsuyasu; Cravatt, Benjamin F; Nagano, Tetsuo

    2013-04-24

    We have established the diced electrophoresis gel (DEG) assay as a proteome-wide screening tool to identify enzymes with activities of interest using turnover-based fluorescent substrates. The method utilizes the combination of native polyacrylamide gel electrophoresis (PAGE) with a multiwell-plate-based fluorometric assay to find protein spots with the specified activity. By developing fluorescent substrates that mimic the structure of neutrophil chemoattractants, we could identify enzymes involved in metabolic inactivation of the chemoattractants.

  18. Multiplex PCR Assay for Identification of Human Diarrheagenic Escherichia coli

    OpenAIRE

    2003-01-01

    A multiplex PCR assay for the identification of human diarrheagenic Escherichia coli was developed. The targets selected for each category were eae for enteropathogenic E. coli, stx for Shiga toxin-producing E. coli, elt and est for enterotoxigenic E. coli, ipaH for enteroinvasive E. coli, and aggR for enteroaggregative E. coli. This assay allowed the categorization of a diarrheagenic E. coli strain in a single reaction tube.

  19. Protein-protein interaction assays: eliminating false positive interactions

    OpenAIRE

    Nguyen, Tuan N.; Goodrich, James A.

    2006-01-01

    Many methods commonly used to identify and characterize interactions between two or more proteins are variations of the immobilized protein-protein interaction assay (for example, glutathione S-transferase (GST) pulldown and coimmunoprecipitation). A potential, and often overlooked, problem with these assays is the possibility that an observed interaction is mediated not by direct contact between proteins, but instead by nucleic acid contaminating the protein preparations. As a negatively cha...

  20. Micronuclei assay: A potential biomonitoring protocol in occupational exposure studies.

    Science.gov (United States)

    Palanikumar, L; Panneerselvam, N

    2011-09-01

    As micronuclei (MN) derive from chromosomal fragments and whole chromosomes lagging behind in anaphase, the MN assay can be used to show both clastogenic and aneugenic effects. This particularly concerns the use of MN as a biomarker ofgenotoxic exposure and effects, where differences in MN frequencies between exposed subjects and referents are expected to be small. The present paper reviews the use of the MN assay in biomonitoring of occupational exposure studies.

  1. Embryotoxicity assays for leached components from dental restorative materials

    OpenAIRE

    Mummolo Stefano; Tecco Simona; Gallusi Gianni; Farini Donatella; Klinger Francesca G; Marzo Giuseppe; Libonati Antonio; De Felici Massimo; Campanella Vincenzo

    2011-01-01

    Abstract Background Currently, there are no suitable assays available to evaluate the embryotoxicity of leached components from restorative dental materials. Methods The effect of the medium conditioned by composites and amalgam on mouse blastocysts in vitro was tested. The materials were also subcutaneously implanted, and the effect of the medium supplemented with serum from the host blood was evaluated in the embryotoxicity assay. The embryo implantation rate in the material-transplanted mo...

  2. Detection of Streptococcus pyogenes using rapid visual molecular assay.

    Science.gov (United States)

    Zhao, Xiangna; He, Xiaoming; Li, Huan; Zhao, Jiangtao; Huang, Simo; Liu, Wei; Wei, Xiao; Ding, Yiwei; Wang, Zhaoyan; Zou, Dayang; Wang, Xuesong; Dong, Derong; Yang, Zhan; Yan, Xiabei; Huang, Liuyu; Du, Shuangkui; Yuan, Jing

    2015-09-01

    Streptococcus pyogenes is an increasingly important pathogen in many parts of the world. Rapid and accurate detection of S. pyogenes aids in the control of the infection. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed and validated for the specific detection of S. pyogenes. The assay incorporates two methods: a chromogenic analysis using a calcein/Mn(2+) complex and real-time turbidity monitoring to assess the reaction. Both methods detected the target DNA within 60 min under 64°C isothermal conditions. The assay used specifically designed primers to target spy1258, and correctly identified 111 strains of S. pyogenes and 32 non-S. pyogenes strains, including other species of the genus Streptococcus. Tests using reference strains showed that the LAMP assay was highly specific. The sensitivity of the assay, with a detection limit of 1.49 pg DNA, was 10-fold greater than that of PCR. The LAMP assay established in this study is simple, fast and sensitive, and does not rely upon any special equipment; thus, it could be employed in clinical diagnosis.

  3. First 25-hydroxyvitamin D assay for general chemistry analyzers.

    Science.gov (United States)

    Saida, Fakhri B; Chen, Xiaoru; Tran, Kiet; Dou, Chao; Yuan, Chong

    2015-03-01

    25-Hydroxyvitamin D [25(OH)D], the predominant circulating form of vitamin D, is an accurate indicator of the general vitamin D status of an individual. Because vitamin D deficiencies have been linked to several pathologies (including osteoporosis and rickets), accurate monitoring of 25(OH)D levels is becoming increasingly important in clinical settings. Current 25(OH)D assays are either chromatographic or immunoassay-based assays. These assays include HPLC, liquid chromatography-tandem mass spectrometry (LC-MS/MS), enzyme-immunosorbent, immunochemiluminescence, immunofluorescence and radioimmunoassay. All these assays use heterogeneous formats that require phase separation and special instrumentations. In this article, we present an overview of these assays and introduce the first homogeneous assay of 25(OH)D for use on general chemistry analyzers. A special emphasis is put on the unique challenges posed by the 25(OH)D analyte. These challenges include a low detection limit, the dissociation of the analyte from its serum transporter and the inactivation of various binding proteins without phase separation steps.

  4. Troponin assays in the assessment of the equine myocardium.

    Science.gov (United States)

    Rossi, T M; Pyle, W G; Maxie, M G; Pearl, D L; Physick-Sheard, P W

    2014-05-01

    In 2000, troponin assays were adopted as the test of choice for detection of myocardial injury in man. This decision was made after extensive testing and followed a 60 year search for a biomarker of myocardial damage with sufficient analytical sensitivity and specificity. This has led to proliferation of assays for use in human medicine, each requiring extensive testing and validation before it could be made available on the open market for human use. The search for ever-more analytically sensitive assays and for a standard reference material continues. The adoption of troponin testing in veterinary medicine followed shortly after its development for use in man, providing a much-needed means of detecting and monitoring myocardial damage in horses. However, application of these tests in veterinary medicine has exclusively involved use of assays designed for and clinically validated in human patients. There is no mandated requirement for test validation in veterinary medicine and, while many of these assays have been shown to be capable of detecting equine troponin, the wide diversity of available tests, lack of validation, absence of protocols for their use and lack of standardisation make their application problematic. The objective of this review article is to address this issue, offering guidance where data are available and encouraging caution where there are none. Ultimately, the overall goal of this review is to examine critically the use of troponin assays in the horse and to promote the accurate and appropriate interpretation of valid results.

  5. Colonization of the meat extracellular matrix proteins by O157 and non-O157 enterohemorrhagic Escherichia coli.

    Science.gov (United States)

    Chagnot, Caroline; Caccia, Nelly; Loukiadis, Estelle; Ganet, Sarah; Durand, Alexandra; Bertin, Yolande; Talon, Régine; Astruc, Thierry; Desvaux, Mickaël

    2014-10-01

    Enterohemorrhagic Escherichia coli (EHEC) are anthropozoonotic agents that range third among food-borne pathogens respective to their incidence and dangerousness in the European Union. EHEC are Shiga-toxin producing E. coli (STEC) responsible for foodborne poisoning mainly incriminated to the consumption of contaminated beef meat. Among the hundreds of STEC serotypes identified, EHEC mainly belong to O157:H7 but non-O157 can represent 20 to 70% of EHEC infections per year. Seven of those serogroups are especially of high-risk for human health, i.e. O26, O45, O103, O111, O121, O145 and O104. While meat can be contaminated all along the food processing chain, EHEC contamination essentially occurs at the dehiding stage of slaughtering. Investigating bacterial colonization to the skeletal-muscle extracellular matrix (ECM) proteins, it appeared that environmental factors influenced specific and non-specific bacterial adhesion of O157 and non-O157 EHEC as well as biofilm formation. Importantly, mechanical treatment (i.e. shaking, centrifugation, pipetting and vortexing) inhibited and biased the results of bacterial adhesion assay. Besides stressing the importance of the protocol to investigate bacterial adhesion to ECM proteins, this study demonstrated that the colonization abilities to ECM proteins vary among EHEC serogroups and should ultimately be taken into consideration to evaluate the risk of contamination for different types of food matrices.

  6. Hemizona Assay and Sperm Penetration Assay in the Prediction of IVF Outcome: A Systematic Review

    Directory of Open Access Journals (Sweden)

    Paraskevi Vogiatzi

    2013-01-01

    Full Text Available The limited predictive value of semen analysis in achieving natural conception or in IVF outcome confirms the need for sperm function tests to determine optimal management. We reviewed HZA and SPA predictive power in IVF outcome, with statistical significance of diagnostic power of the assays. HZA was readily efficient in predicting IVF outcome, while evident inconsistency among the studies analysed framed the SPA’s role in male fertility evaluation. Considerable variation was noted in the diagnostic accuracy values of SPA with wide sensitivity (52–100%, specificity (0–100%, and PPV (18–100% and NPV (0–100% together with fluctuation and notable differentiation in methodology and cutoff values employed by each group. HZA methodology was overall consistent with minor variation in cutoff values and oocyte source, while data analysis reported strong correlation between HZA results with IVF outcome, high sensitivity (75–100%, good specificity (57–100%, and high PPV (79–100% and NPV (68–100%. HZA correlated well with IVF outcome and demonstrated better sensitivity/specificity and positive/negative predictive power. Males with normal or slightly abnormal semen profiles could benefit by this intervention and could be evaluated prior to referral to assisted reproduction. HZA should be used in a sequential fashion with semen analysis and potentially other bioassays in an IVF setting.

  7. Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture.

    Science.gov (United States)

    Cory, A H; Owen, T C; Barltrop, J A; Cory, J G

    1991-07-01

    A new tetrazolium analog of 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) was evaluated as a substitute for MTT in the microculture screening assay for in vitro cell growth. This new tetrazolium compound, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2- (4-sulfophenyl)-2H-tetrazolium, inner salt (MTS), in the presence of phenazine methosulfate (PMS), gave a water-soluble formazan product that had an absorbance maximum at 490-500 nm in phosphate-buffered saline. The amount of colored product formed was proportional to the number of cells and the time of incubation of the cells with MTS/PMS. MTS/PMS was reactive in all the cell lines tested which included mouse leukemia L1210 cells, mouse Ehrlich tumor cells, mouse 3T3 fibroblasts, and human colon tumor cells (HT-29). HT-29 and 3T3 fibroblasts reduced MTS/PMS more efficiently than they reduced MTT. Comparable to the amount of product formed from MTT, MTS/PMS gave excellent product formation. The IC50 value for pyrazoloimidazole obtained using MTS/PMS was 200 microM; for 5-fluoro-2'-deoxyuridine, the IC50 value was 0.9 nM. These values compared very favorably with the IC50 values obtained by direct cell counts. Further, the same IC50 values were obtained when the absorbances of the formazan product in the 96-well plates were determined after different times of incubation.

  8. Proceedings of the 2010 AFMS Medical Research Symposium. Volume 1. Plenary Sessions, Presentation and Poster Abstracts

    Science.gov (United States)

    2011-03-15

    0157:H7 (EHEC), Shigella, Salmonella, and Campylobacter , and evaluated using both the HMLK and the JBAIDS species- specific assays. RESULTS: EHEC...detected by HMLK but detected by JBAIDS. Campylobacter was not detected at high or low concentrations by the HMLK but was successfully detected by...cMAbs stably produced by the DHFR double mutant Chinese hamster ovarian (CHO)- DG44 cells were affinity-purified. By ELISA and Western blot analysis

  9. A colorimetric sandwich-type assay for sensitive thrombin detection based on enzyme-linked aptamer assay.

    Science.gov (United States)

    Park, Jun Hee; Cho, Yea Seul; Kang, Sungmuk; Lee, Eun Jeong; Lee, Gwan-Ho; Hah, Sang Soo

    2014-10-01

    A colorimetric sandwich-type assay based on enzyme-linked aptamer assay has been developed for the fast and sensitive detection of as low as 25 fM of thrombin with high linearity. Aptamer-immobilized glass was used to capture the target analyte, whereas a second aptamer, functionalized with horseradish peroxidase (HRP), was employed for the conventional 3,5,3',5'-tetramethylbenzidine (TMB)-based colorimetric detection. Without the troublesome antibody requirement of the conventional enzyme-linked immunosorbent assay (ELISA), as low as 25 fM of thrombin could be rapidly and reproducibly detected. This assay has superior, or at least equal, recovery and accuracy to that of conventional antibody-based ELISA.

  10. The Granzyme B ELISPOT assay: an alternative to the 51Cr-release assay for monitoring cell-mediated cytotoxicity

    Directory of Open Access Journals (Sweden)

    Baseler Michael

    2003-12-01

    Full Text Available Abstract Background The interferon-γ (IFN-γ ELISPOT assay is one of the most useful techniques for immunological monitoring of cancer vaccine trials and has gained increased application as a measure of specific T cell activation. However, it does not assess cell-mediated cytotoxicity directly as IFN-γ secretion is not limited to only cytolytic cells. Granzyme B (GrB is a key mediator of target cell death via the granule-mediated pathway. Therefore, the release of GrB by cytolytic lymphocytes upon effector-target interaction may be a more specific indicator of CTL and NK cytotoxic ability than IFN-γ secretion. Methods We assessed whether the GrB ELISPOT assay is a viable alternative to the 51Cr-release and IFN-γ ELISPOT assays for measuring antigen-specific CTL cytotoxicity. Direct comparisons between the three assays were made using human CTL cell lines (αEN-EBV and αJY and an in vitro stimulated anti-Flu matrix peptide (FMP-specific CTL. Results When the GrB ELISPOT was directly compared to the IFN-γ ELISPOT and 51Cr-release assays, excellent cross-correlation between all three assays was shown. However, measurable IFN-γ secretion in the ELISPOT assay was observed only after 1 hour of incubation and cytotoxicity assessed via the 51Cr-release assay after 4 hours, whereas GrB secretion was detectable within 10 min of effector-target contact with significant secretion observed after 1 h. Titration studies demonstrated a strong correlation between the number of effector cells and GrB spots per well. Irrelevant targets or antigens did not induce significant GrB secretion. Additionally, GrB secretion was abrogated when CTL cultures were depleted of CD8+ cells. Conclusion Our findings demonstrate that the GrB ELISPOT assay is a superior alternative to the 51Cr-release assay since it is significantly more sensitive and provides an estimation of cytotoxic effector cell frequency. Additionally, unlike the IFN-γ ELISPOT assay, the GrB ELISPOT

  11. Evaluation of Presto(plus) assay and LightMix kit Trichomonas vaginalis assay for detection of Trichomonas vaginalis in dry vaginal swabs.

    Science.gov (United States)

    de Waaij, Dewi J; Ouburg, Sander; Dubbink, Jan Henk; Peters, Remco P H; Morré, Servaas A

    2016-08-01

    This is an evaluation study of the Presto(plus) Assay for T. vaginalis by comparing to the TIB MOLBIOL LightMix Kit Trichomonas vaginalis Assay using 615 dry collected vaginal and rectal swabs. Discordant samples were analyzed by the Qiagen® Microbial DNA qPCR for TV Assay. Both assays showed comparable performances (McNemar p>0.05).

  12. Suitability of a liquid chromatography assay of neomycin sulfate to replace the microbiological assay for neomycin in USP Monographs.

    Science.gov (United States)

    Hanko, Valoran P; Rohrer, Jeffrey S

    2010-01-01

    The current USP National Formulary contains 65 Monographs for drug formulations containing neomycin. All 65 Monographs prescribe a bioassay for neomycin assay. This bioassay, based on cell culture, is labor intensive, has poor precision, and cannot be adapted for purity or identification. High-performance anion-exchange chromatography with integrated pulsed amperometric detection (HPAE-IPAD), a liquid chromatography technique, has been shown to be suitable for neomycin purity analysis and neomycin assay of an over-the-counter first aid cream (Hanko and Rohrer [17]). Here we propose that an HPAE-IPAD assay can replace the bioassay in the 65 neomycin-containing Monographs. We applied the HPAE-IPAD assay to four neomycin-containing drug products representing the four classes of formulations found in the 65 Monographs, liquid, solid, suspension, and cream. Each drug was analyzed with two chromatography systems, and on 3 separate days. For all products, HPAE-IPAD measurements were precise and accurate with respect to the label concentrations. There was also high accuracy for spike recovery of neomycin from the four drug products throughout 70-150% of the labeled concentration. These results suggest that an HPAE-IPAD assay would be an accurate assay for neomycin, and would be faster and more precise than the current bioassay.

  13. Neutron Resonance Transmission Analysis (NRTA): A Nondestructive Assay Technique for the Next Generation Safeguards Initiative’s Plutonium Assay Challenge

    Energy Technology Data Exchange (ETDEWEB)

    J. W. Sterbentz; D. L. Chichester

    2010-12-01

    This is an end-of-year report for a project funded by the National Nuclear Security Administration's Office of Nuclear Safeguards (NA-241). The goal of this project is to investigate the feasibility of using Neutron Resonance Transmission Analysis (NRTA) to assay plutonium in commercial light-water-reactor spent fuel. This project is part of a larger research effort within the Next-Generation Safeguards Initiative (NGSI) to evaluate methods for assaying plutonium in spent fuel, the Plutonium Assay Challenge. The first-year goals for this project were modest and included: 1) developing a zero-order MCNP model for the NRTA technique, simulating data results presented in the literature, 2) completing a preliminary set of studies investigating important design and performance characteristics for the NRTA measurement technique, and 3) documentation of this work in an end of the year report (this report). Research teams at Los Alamos National Laboratory (LANL), Lawrence Berkeley National Laboratory (LBNL), Pacific Northwest National Laboratory (PNNL), and at several universities are also working to investigate plutonium assay methods for spent-fuel safeguards. While the NRTA technique is well proven in the scientific literature for assaying individual spent fuel pins, it is a newcomer to the current NGSI efforts studying Pu assay method techniques having just started in March 2010; several analytical techniques have been under investigation within this program for two to three years or more. This report summarizes a nine month period of work.

  14. Sperm DNA assays and their relationship to sperm motility and morphology in bulls (Bos Taurus).

    Science.gov (United States)

    Serafini, Rosanna; Romano, Juan E; Varner, Dickson D; Di Palo, Rossella; Love, Charles C

    2015-08-01

    The relationship among sperm DNA assays in bulls with different sperm motility and morphology measures has not been reported. The objectives of the present study were to (1) describe Comet assay measures and examine their repeatability (inter- and intra-assay); (2) compare sperm DNA quality assays (i.e., Sperm Chromatin Structure Assay-SCSA; alkaline and neutral Comet assays and Sperm Bos Halomax assay-SBH) in two groups of bulls selected on either greater and lesser sperm motility and morphology (greater compared with lesser); (3) determine the relationship among DNA assays and sperm motility and morphology values. Inter-assay repeatability was greater for the neutral Comet assay as compared to the alkaline Comet assay. Intra-assay repeatability was greater than inter-assay repeatability for both Comet assays. Comet assay dimension measures and percentage tail DNA were the most repeatable for both Comet assays. Among sperm DNA quality assays, only SCSA measures and neutral Comet assay Ghosts (% Ghosts), head diameter and area, and comet area were different between greater and lesser sperm quality groups (P<0.05). The SCSA measures were inversely correlated with neutral Comet head measures (diameter, area, and intensity) and positively with percentage Ghosts (P<0.05). The % Ghosts and COMP-αt were correlated with some measures of sperm morphology and sperm motility. The neutral Comet assay was more appropriate for sperm evaluation than the alkaline Comet assay for distinguishing among groups with different sperm quality.

  15. A versatile polyacrylamide gel electrophoresis based sulfotransferase assay

    Directory of Open Access Journals (Sweden)

    Prather Brittany

    2010-02-01

    Full Text Available Abstract Background Sulfotransferases are a large group of enzymes that regulate the biological activity or availability of a wide spectrum of substrates through sulfation with the sulfur donor 3'-phosphoadenosine-5'-phosphosulfate (PAPS. These enzymes are known to be difficult to assay. A convenient assay is needed in order to better understand these enzymes. Results A universal sulfotransferase assay method based on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE is described. This assay has been successfully applied to substrates as small as α-naphthol and as big as proteoglycans. As examples, we present the assays for recombinant human CHST4, TPST1, CHST3 and HS6ST1. In order to assess whether a small molecule can be applicable to this type of assay, a method to estimate the relative mobility of a molecule to PAPS is also presented. The estimated relative mobilities of various sulfated small molecules generated by SULT1A1, SULT1E1, SULT2A1 and CHST4 are in the range of ± 0.2 of the actual relative mobilities. Conclusion The versatility of the current method comes from the ability that SDS-PAGE can separate proteins and small molecules according to different parameters. While mobilities of proteins during SDS-PAGE are inversely related to their sizes, mobilities of small molecules are positively related to their charge/mass ratios. The predicted relative mobility of a product to PAPS is a good indicator of whether a sulfotransferase can be assayed with SDS-PAGE. Because phosphorylation is most similar to sulfation in chemistry, the method is likely to be applicable to kinases as well.

  16. Recommendations for safety testing with the in vivo comet assay.

    Science.gov (United States)

    Vasquez, Marie Z

    2012-08-30

    While the in vivo comet assay increases its role in regulatory safety testing, deliberations about the interpretation of comet data continue. Concerns can arise regarding comet assay publications with limited data from non-blind testing of positive control compounds and using protocols (e.g. dose concentrations, sample times, and tissues) known to give an expected effect. There may be a tendency towards bias when the validation or interpretation of comet assay data is based on results generated by widely accepted but non-validated assays. The greatest advantages of the comet assay are its sensitivity and its ability to detect genotoxicity in tissues and at sample times that could not previously be evaluated. Guidelines for its use and interpretation in safety testing should take these factors into account. Guidelines should be derived from objective review of data generated by blind testing of unknown compounds dosed at non-toxic concentrations and evaluated in a true safety-testing environment, where the experimental design and conclusions must be defensible. However, positive in vivo comet findings with such compounds are rarely submitted to regulatory agencies and this data is typically unavailable for publication due to its proprietary nature. To enhance the development of guidelines for safety testing with the comet assay, and with the permission of several sponsors, this paper presents and discusses relevant data from multiple GLP comet studies conducted blind, with unknown pharmaceuticals and consumer products. Based on these data and the lessons we have learned through the course of conducting these studies, I suggest significant adjustments to the current conventions, and I provide recommendations for interpreting in vivo comet assay results in situations where risk must be evaluated in the absence of carcinogenicity or clinical data.

  17. Worldwide interest in the comet assay: a bibliometric study.

    Science.gov (United States)

    Neri, Monica; Milazzo, Daniele; Ugolini, Donatella; Milic, Mirta; Campolongo, Alessandra; Pasqualetti, Patrizio; Bonassi, Stefano

    2015-01-01

    The comet assay is a rapid, sensitive and relatively simple method for measuring DNA damage. A bibliometric study was performed to evaluate temporal and geographical trends, research quality and main areas of interest in scientific production in this field. A PubMed search strategy was developed and 7674 citations were retrieved in the period 1990-2013. Notably, the MeSH (Medical Subject Headings) term 'comet assay', officially introduced in 2000, is used by indexers only in two thirds of papers retrieved. Articles on the comet assay were published in 78 countries, spread over the 5 continents. The EU contributed the greatest output, producing >2900 articles with IF (42.0%) and totalling almost 10000 IF points, and was followed by USA. In the new millennium, research with this assay reached a plateau or slow decline in the most industrialised areas (USA, Germany, UK, Italy), while its use has boomed in emerging countries, with increases of 5- to 7-fold in the last 10 years in China, India and Brazil, for instance. This transition resulted in a slow decrease of scientific production quality, as the countries that increased their relative weight typically had lower mIFs. The most common MeSH terms used in papers using the comet assay referred to wide areas of interest, such as DNA damage and repair, cell survival and apoptosis, cancer and oxidative stress, occupational and environmental health. Keywords related to humans, rodents and cell culture were also frequently used. The top journal for the comet assay articles was found to be Mutation Research, followed by Mutagenesis. Most papers using the comet assay as a biomarker were published in genetic and toxicology journals, with a stress on environmental and occupational disciplines.

  18. Evolving BioAssay Ontology (BAO): modularization, integration and applications.

    Science.gov (United States)

    Abeyruwan, Saminda; Vempati, Uma D; Küçük-McGinty, Hande; Visser, Ubbo; Koleti, Amar; Mir, Ahsan; Sakurai, Kunie; Chung, Caty; Bittker, Joshua A; Clemons, Paul A; Brudz, Steve; Siripala, Anosha; Morales, Arturo J; Romacker, Martin; Twomey, David; Bureeva, Svetlana; Lemmon, Vance; Schürer, Stephan C

    2014-01-01

    The lack of established standards to describe and annotate biological assays and screening outcomes in the domain of drug and chemical probe discovery is a severe limitation to utilize public and proprietary drug screening data to their maximum potential. We have created the BioAssay Ontology (BAO) project (http://bioassayontology.org) to develop common reference metadata terms and definitions required for describing relevant information of low-and high-throughput drug and probe screening assays and results. The main objectives of BAO are to enable effective integration, aggregation, retrieval, and analyses of drug screening data. Since we first released BAO on the BioPortal in 2010 we have considerably expanded and enhanced BAO and we have applied the ontology in several internal and external collaborative projects, for example the BioAssay Research Database (BARD). We describe the evolution of BAO with a design that enables modeling complex assays including profile and panel assays such as those in the Library of Integrated Network-based Cellular Signatures (LINCS). One of the critical questions in evolving BAO is the following: how can we provide a way to efficiently reuse and share among various research projects specific parts of our ontologies without violating the integrity of the ontology and without creating redundancies. This paper provides a comprehensive answer to this question with a description of a methodology for ontology modularization using a layered architecture. Our modularization approach defines several distinct BAO components and separates internal from external modules and domain-level from structural components. This approach facilitates the generation/extraction of derived ontologies (or perspectives) that can suit particular use cases or software applications. We describe the evolution of BAO related to its formal structures, engineering approaches, and content to enable modeling of complex assays and integration with other ontologies and

  19. Epithelial cells as alternative human biomatrices for comet assay

    Directory of Open Access Journals (Sweden)

    Emilio eRojas

    2014-11-01

    Full Text Available The comet assay is a valuable experimental tool aimed at mapping DNA damage in human cells in vivo for environmental and occupational monitoring, as well as for therapeutic purposes, such as storage prior to transplant, during tissue engineering, and in experimental ex vivo assays. Furthermore, due to its great versatility, the comet assay allows to explore the use of alternative cell types to assess DNA damage, such as epithelial cells. Epithelial cells, as specialized components of many organs, have the potential to serve as biomatrices that can be used to evaluate genotoxicity and may also serve as early effect biomarkers. Furthermore, 80% of solid cancers are of epithelial origin, which points to the importance of studying DNA damage in these tissues. Indeed, studies including comet assay in epithelial cells have either clear clinical applications (lens and corneal epithelial cells or examine genotoxicity within human biomonitoring and in vitro studies. We here review improvements in determining DNA damage using the comet assay by employing lens, corneal, tear duct, buccal, and nasal epithelial cells. For some of these tissues invasive sampling procedures are needed. Desquamated epithelial cells must be obtained and dissociated prior to examination using the comet assay, and such procedures may induce varying amounts of DNA damage. Buccal epithelial cells require lysis enriched with proteinase K to obtain free nucleosomes.Over a thirty year period, the comet assay in epithelial cells has been litlle employed, however its use indicates that it could be an extraordinary tool not only for risk assessment, but also for diagnosis, prognosis of treatments and diseases.

  20. Multiplexed Recombinase Polymerase Amplification Assay To Detect Intestinal Protozoa.

    Science.gov (United States)

    Crannell, Zachary; Castellanos-Gonzalez, Alejandro; Nair, Gayatri; Mejia, Rojelio; White, A Clinton; Richards-Kortum, Rebecca

    2016-02-01

    This work describes a proof-of-concept multiplex recombinase polymerase amplification (RPA) assay with lateral flow readout that is capable of simultaneously detecting and differentiating DNA from any of the diarrhea-causing protozoa Giardia, Cryptosporidium, and Entamoeba. Together, these parasites contribute significantly to the global burden of diarrheal illness. Differential diagnosis of these parasites is traditionally accomplished via stool microscopy. However, microscopy is insensitive and can miss up to half of all cases. DNA-based diagnostics such as polymerase chain reaction (PCR) are far more sensitive; however, they rely on expensive thermal cycling equipment, limiting their availability to centralized reference laboratories. Isothermal DNA amplification platforms, such as the RPA platform used in this study, alleviate the need for thermal cycling equipment and have the potential to broaden access to more sensitive diagnostics. Until now, multiplex RPA assays have not been developed that are capable of simultaneously detecting and differentiating infections caused by different pathogens. We developed a multiplex RPA assay to detect the presence of DNA from Giardia, Cryptosporidium, and Entamoeba. The multiplex assay was characterized using synthetic DNA, where the limits-of-detection were calculated to be 403, 425, and 368 gene copies per reaction of the synthetic Giardia, Cryptosporidium, and Entamoeba targets, respectively (roughly 1.5 orders of magnitude higher than for the same targets in a singleplex RPA assay). The multiplex assay was also characterized using DNA extracted from live parasites spiked into stool samples where the limits-of-detection were calculated to be 444, 6, and 9 parasites per reaction for Giardia, Cryptosporidium, and Entamoeba parasites, respectively. This proof-of-concept assay may be reconfigured to detect a wide variety of targets by re-designing the primer and probe sequences.

  1. Comparison of clonogenic assay with premature chromosome condensation assay in prediction of human cell radiosensitivity

    Institute of Scientific and Technical Information of China (English)

    Zhuan-Zi Wang; Wen-Jian Li; Hong Zhang; Jian-She Yang; Rong Qiu; Xiao Wang

    2006-01-01

    AIM: To determine whether the number of non-rejoining G2-chromatid breaks can predict the radiosensitivity of human cell lines.METHODS: Cell lines of human ovary carcinoma cells (HO8910), human hepatoma cells (HepG2) and liver cells (L02) were irradiated with a range of doses and assessed both of cell survival and non-rejoining G2-chromatid breaks at 24 h after irradiation. Cell survival was documented by a colony assay. Non-rejoining G2-chromatid breaks were measured by counting the number of non-rejoining G2 chromatid breaks at 24 h after irradiation, detected by the prematurely chromosome condensed (PCC) technique.RESULTS: A linear-quadratic survival curve was observed in three cell lines, and HepG2 was the most sensitive to y-radiation. A dose-dependent linear increase was observed in radiation-induced non-rejoining G2-PCC breaks measured at 24 h after irradiation in all cell lines, and HepG2 was the most susceptible to induction of non-rejoining G2-PCC breaks. A close correlation was found between the clonogenic radiosensitivity and the radiation-induced non-rejoining G2-PCC breaks (r= 0.923). Furthermore, survival-aberration correlations for two or more than two doses lever were also significant.CONCLUSION: The number of non-rejoining G2 PCC breaks holds considerable promise for predicting the radiosensitivity of normal and tumor cells when two or more than two doses lever is tested.

  2. The comet assay in testing the potential genotoxicity of nanomaterials

    Directory of Open Access Journals (Sweden)

    Amaya Azqueta

    2015-06-01

    Full Text Available In the last two decades the production and use of nanomaterials (NMs has impressively increased. Their small size, given a mass equal to that of the corresponding bulk material, implies an increase in the surface area and consequently in the number of atoms that can be reactive. They possess different physical, chemical and biological properties compared to bulk materials of the same composition, which makes them very interesting and valuable for many different applications in technology, energy, construction, electronics, agriculture, optics, paints, textiles, food, cosmetics, medicine... Toxicological assessment of NMs is crucial; the same properties that make them interesting also make them potentially harmful for health and the environment. However, the term NM covers many different kinds of particle , and so there is no simple, standard approach to assessing their toxicity. NMs can enter the cell, interact with cell components and even penetrate the nucleus and interfere with the genetic material. Among the different branches of toxicology, genotoxicity is a main area of concern since it is closely related with the carcinogenic potential of compounds. The Organisation for Economic Co-operation and Development (OECD has published internationally agreed in vitro and in vivo validated test methods to evaluate different genotoxic endpoints of chemicals, including chromosome and gene mutations, and DNA breaks. However not all the assays are suitable to study the genotoxic potential of NMs as has been shown by the OECD Working Party on Manufactured Nanomaterials (WPMN. Moreover, alterations to DNA bases, which are precursors to mutations and of great importance in elucidating the mechanism of action of NMs, are not covered by the OECD guidelines. The in vivo standard comet assay (which measures DNA breaks and alkali-labile sites was included in the OECD assays battery in September 2014 while the in vitro standard comet assay is currently under

  3. Quantitative CrAssphage PCR Assays for Human Fecal ...

    Science.gov (United States)

    Environmental waters are monitored for fecal pollution to protect public health and water resources. Traditionally, general fecal indicator bacteria are used; however, they cannot distinguish human fecal waste from pollution from other animals. Recently, a novel bacteriophage, crAssphage, was discovered by metagenomic data mining and reported to be abundant in and closely associated with human fecal waste. To confirm bioinformatic predictions, 384 primer sets were designed along the length of the crAssphage genome. Based upon initial screening, two novel crAssphage qPCR assays (CPQ_056 and CPQ_064) were designed and evaluated in reference fecal samples and water matrices. The assays exhibited high specificities (98.6%) when tested against a large animal fecal reference library and were highly abundant in raw sewage and sewage impacted water samples. In addition, CPQ_056 and CPQ_064 assay performance was compared to HF183/BacR287 and HumM2 methods in paired experiments. Findings confirm viral crAssphage qPCR assays perform at a similar level to well established bacterial human-associated fecal source identification technologies. These new viral based assays could become important water quality management and research tools. To inform the public.

  4. A radioactive assay for the degradation of neuropeptide Y

    Energy Technology Data Exchange (ETDEWEB)

    Ludwig, R.; Lucius, R.; Mentlein, R. [Kiel Univ. (Germany)

    1995-12-31

    Neuropeptide Y (NPY) is one of the most abundant neuropeptides in the mammalian central nervous system. Like other neuropeptides, NPY is inactivated by specialized neuro-peptidases. To trace the degradation of NPY, an assay was established using biotinylated NPY. Biotinyl-NPY was radiolabeled with Na{sup 125}I by the chloramine-T method and bound to a streptavidin-agarose matrix. The amount of radiolabeling was analyzed by reverse-phase HPLC. The assay was carried out with five peptidases and inhibitors to demonstrate different specific activity. Measurable amounts of radioactivity were released by treatment with endopeptidase-24.18, plasmin, and trypsin, whereas dipetidylpeptidase IV (DPPIV) and angiotensin-converting enzyme (ACE) showed no activity in this assay. In the case of DPPIV this is due to a resistance of the assay to aminopeptidase attack. The assay is useful to study the specific degradation of NPY particularly by endopeptidases in all kinds of biological samples. (authors). 31 refs., 6 figs.

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

    Directory of Open Access Journals (Sweden)

    Conceição LV Santos

    2015-06-01

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

  6. Non-Destructive Assay of Curium Contaminated Transuranic Waste Drums

    Energy Technology Data Exchange (ETDEWEB)

    Foster, L.A.

    1998-11-01

    At the Plutonium Facility at Los Alamos National Laboratory, a series of non-destructive assays were performed on five transuranic waste (TRU) drums containing non-plutonium scrap metal that was potentially contaminated with weapons grade plutonium and trace quantities of curium. Typically, waste drums containing metal matrices are assayed for plutonium content using passive neutron coincidence counting techniques. The presence of trace quantities of Cm-244 prevents this type of analysis because of the strong coincidence signal created by spontaneous fission of Cm-244. To discriminate between the plutonium and curium materials present, an active neutron measurement technique was used. A Cf shuffler designed for measurement of uranium bearing materials was calibrated for plutonium in the active mode. The waste drums were then assayed for plutonium content in the shuffler using the active-mode calibration. The curium contamination levels were estimated from the difference between the active-mode measurement in the shuffler and a passive assay in a neutron coincidence counter. Far field gamma-ray measurements were made to identify additional radioactive contaminants and to corroborate the plutonium measurement results obtained from the active-mode assay. This report describes in detail the measurement process used for characterization of these waste drums. The measurement results and the estimated uncertainty will be presented.

  7. Capture-stabilize approach for membrane protein SPR assays.

    Science.gov (United States)

    Chu, Ruiyin; Reczek, David; Brondyk, William

    2014-12-08

    Measuring the binding kinetics of antibodies to intact membrane proteins by surface plasmon resonance has been challenging largely because of the inherent difficulties in capturing membrane proteins on chip surfaces while retaining their native conformation. Here we describe a method in which His-tagged CXCR5, a GPCR, was purified and captured on a Biacore chip surface via the affinity tag. The captured receptor protein was then stabilized on the chip surface by limited cross-linking. The resulting chip surface retained ligand binding activity and was used for monoclonal antibody kinetics assays by a standard Biacore kinetics assay method with a simple low pH regeneration step. We demonstrate the advantages of this whole receptor assay when compared to available peptide-based binding assays. We further extended the application of the capture-stabilize approach to virus-like particles and demonstrated its utility analyzing antibodies against CD52, a GPI-anchored protein, in its native membrane environment. The results are the first demonstration of chemically stabilized chip surfaces for membrane protein SPR assays.

  8. A highly scalable peptide-based assay system for proteomics.

    Directory of Open Access Journals (Sweden)

    Igor A Kozlov

    Full Text Available We report a scalable and cost-effective technology for generating and screening high-complexity customizable peptide sets. The peptides are made as peptide-cDNA fusions by in vitro transcription/translation from pools of DNA templates generated by microarray-based synthesis. This approach enables large custom sets of peptides to be designed in silico, manufactured cost-effectively in parallel, and assayed efficiently in a multiplexed fashion. The utility of our peptide-cDNA fusion pools was demonstrated in two activity-based assays designed to discover protease and kinase substrates. In the protease assay, cleaved peptide substrates were separated from uncleaved and identified by digital sequencing of their cognate cDNAs. We screened the 3,011 amino acid HCV proteome for susceptibility to cleavage by the HCV NS3/4A protease and identified all 3 known trans cleavage sites with high specificity. In the kinase assay, peptide substrates phosphorylated by tyrosine kinases were captured and identified by sequencing of their cDNAs. We screened a pool of 3,243 peptides against Abl kinase and showed that phosphorylation events detected were specific and consistent with the known substrate preferences of Abl kinase. Our approach is scalable and adaptable to other protein-based assays.

  9. Evaluation of a radioreceptor assay for TSH receptor autoantibodies

    Energy Technology Data Exchange (ETDEWEB)

    Rootwelt, K.

    1988-02-01

    A commercial radioreceptor assay for TSH receptor autoantibodies (TRAb), based on solubilized porcine receptor and purified radio-iodinated bovine TSH, was tested in 264 subjects with a variety of thyroid disorders. The sensitivity of the assay for the detection of hyperthyroid Graves' disease was 91%. The assay specificity for Graves' disease was 95%. With the exception of one patient with Hashimoto's disease and one patient with de Quervain's subacute thyroiditis no subjects other than Graves' patients had detectable TRAb. Thus purely blocking TSII receptor autoantibodies were not detected with the assay. One female with thyroxine-treated idiopathic primary hypothyroidism who had given birth to two children with transiently elevated TSH, was found to have a circulating TSH-binding substance that resulted in an abnormally negative TRAb value, and highly discrepant results when TSH was measured with a double antibody TSH radioimmunoassay and an immunoradiometric assay. The TSH-binding substance was precipitated like a protein, but was not IgG. Similar findings have not previously been reported.

  10. Formalization, annotation and analysis of diverse drug and probe screening assay datasets using the BioAssay Ontology (BAO.

    Directory of Open Access Journals (Sweden)

    Uma D Vempati

    Full Text Available Huge amounts of high-throughput screening (HTS data for probe and drug development projects are being generated in the pharmaceutical industry and more recently in the public sector. The resulting experimental datasets are increasingly being disseminated via publically accessible repositories. However, existing repositories lack sufficient metadata to describe the experiments and are often difficult to navigate by non-experts. The lack of standardized descriptions and semantics of biological assays and screening results hinder targeted data retrieval, integration, aggregation, and analyses across different HTS datasets, for example to infer mechanisms of action of small molecule perturbagens. To address these limitations, we created the BioAssay Ontology (BAO. BAO has been developed with a focus on data integration and analysis enabling the classification of assays and screening results by concepts that relate to format, assay design, technology, target, and endpoint. Previously, we reported on the higher-level design of BAO and on the semantic querying capabilities offered by the ontology-indexed triple store of HTS data. Here, we report on our detailed design, annotation pipeline, substantially enlarged annotation knowledgebase, and analysis results. We used BAO to annotate assays from the largest public HTS data repository, PubChem, and demonstrate its utility to categorize and analyze diverse HTS results from numerous experiments. BAO is publically available from the NCBO BioPortal at http://bioportal.bioontology.org/ontologies/1533. BAO provides controlled terminology and uniform scope to report probe and drug discovery screening assays and results. BAO leverages description logic to formalize the domain knowledge and facilitate the semantic integration with diverse other resources. As a consequence, BAO offers the potential to infer new knowledge from a corpus of assay results, for example molecular mechanisms of action of perturbagens.

  11. A simple high-content cell cycle assay reveals frequent discrepancies between cell number and ATP and MTS proliferation assays.

    Directory of Open Access Journals (Sweden)

    Grace Ka Yan Chan

    Full Text Available In order to efficiently characterize both antiproliferative potency and mechanism of action of small molecules targeting the cell cycle, we developed a high-throughput image-based assay to determine cell number and cell cycle phase distribution. Using this we profiled the effects of experimental and approved anti-cancer agents with a range mechanisms of action on a set of cell lines, comparing direct cell counting versus two metabolism-based cell viability/proliferation assay formats, ATP-dependent bioluminescence, MTS (3-(4,5-dimethylthiazol-2-yl-5-(3-carboxymethoxyphenyl-2-(4-sulfophenyl-2H-tetrazolium reduction, and a whole-well DNA-binding dye fluorescence assay. We show that, depending on compound mechanisms of action, the metabolism-based proxy assays are frequently prone to 1 significant underestimation of compound potency and efficacy, and 2 non-monotonic dose-response curves due to concentration-dependent phenotypic 'switching'. In particular, potency and efficacy of DNA synthesis-targeting agents such as gemcitabine and etoposide could be profoundly underestimated by ATP and MTS-reduction assays. In the same image-based assay we showed that drug-induced increases in ATP content were associated with increased cell size and proportionate increases in mitochondrial content and respiratory flux concomitant with cell cycle arrest. Therefore, differences in compound mechanism of action and cell line-specific responses can yield significantly misleading results when using ATP or tetrazolium-reduction assays as a proxy for cell number when screening compounds for antiproliferative activity or profiling panels of cell lines for drug sensitivity.

  12. Cytogenetic status of healthy children assessed with the alkaline comet assay and the cytokinesis-block micronucleus cytome assay.

    Science.gov (United States)

    Gajski, Goran; Gerić, Marko; Oreščanin, Višnja; Garaj-Vrhovac, Vera

    2013-01-20

    In the present study the alkaline comet assay and the cytokinesis-block micronucleus cytome (CBMN Cyt) assay were used to evaluate the baseline frequency of cytogenetic damage in peripheral blood lymphocytes (PBLs) of 50 healthy children from the general population in Croatia (age, 11.62±1.81 years). Mean values of tail length, tail intensity and tail moment, as comet assay parameters, were 12.92±0.10, 0.73±0.06 and 0.08±0.01, respectively. The mean frequency of micronuclei (MN) for all subjects was 2.32±0.28 per 1000 bi-nucleated cells, while the mean frequency of nucleoplasmic bridges (NPBs) was 1.72±0.24 and of nuclear buds (NBUDs) 1.44±0.19. The mean nuclear division index (NDI) was 1.70±0.05. When comet-assay parameters were considered, higher mean values for all three were found for the female population. According to the Mann-Whitney U test applied on the results of the comet assay, the only statistically significant difference between the male and female populations was found for tail length. Similar to the results obtained by the comet assay, girls showed higher mean values of all three measured parameters of the CBMN Cyt assay. This difference was statistically significant for total number of NPBs only. In the case of the NDI, a higher mean value was also obtained in girls, but this difference was not statistically significant. The results obtained present background data that could be considered as normal values for healthy children living in urban areas, and can later on serve as baseline values for further toxicological monitoring. Additionally, the usefulness of both techniques in measuring cytogenetic damage during bio-monitoring of children is confirmed.

  13. Formalization, annotation and analysis of diverse drug and probe screening assay datasets using the BioAssay Ontology (BAO).

    Science.gov (United States)

    Vempati, Uma D; Przydzial, Magdalena J; Chung, Caty; Abeyruwan, Saminda; Mir, Ahsan; Sakurai, Kunie; Visser, Ubbo; Lemmon, Vance P; Schürer, Stephan C

    2012-01-01

    Huge amounts of high-throughput screening (HTS) data for probe and drug development projects are being generated in the pharmaceutical industry and more recently in the public sector. The resulting experimental datasets are increasingly being disseminated via publically accessible repositories. However, existing repositories lack sufficient metadata to describe the experiments and are often difficult to navigate by non-experts. The lack of standardized descriptions and semantics of biological assays and screening results hinder targeted data retrieval, integration, aggregation, and analyses across different HTS datasets, for example to infer mechanisms of action of small molecule perturbagens. To address these limitations, we created the BioAssay Ontology (BAO). BAO has been developed with a focus on data integration and analysis enabling the classification of assays and screening results by concepts that relate to format, assay design, technology, target, and endpoint. Previously, we reported on the higher-level design of BAO and on the semantic querying capabilities offered by the ontology-indexed triple store of HTS data. Here, we report on our detailed design, annotation pipeline, substantially enlarged annotation knowledgebase, and analysis results. We used BAO to annotate assays from the largest public HTS data repository, PubChem, and demonstrate its utility to categorize and analyze diverse HTS results from numerous experiments. BAO is publically available from the NCBO BioPortal at http://bioportal.bioontology.org/ontologies/1533. BAO provides controlled terminology and uniform scope to report probe and drug discovery screening assays and results. BAO leverages description logic to formalize the domain knowledge and facilitate the semantic integration with diverse other resources. As a consequence, BAO offers the potential to infer new knowledge from a corpus of assay results, for example molecular mechanisms of action of perturbagens.

  14. Qualification of standard membrane-feeding assay with Plasmodium falciparum malaria and potential improvements for future assays.

    Directory of Open Access Journals (Sweden)

    Kazutoyo Miura

    Full Text Available Vaccines that interrupt malaria transmission are of increasing interest and a robust functional assay to measure this activity would promote their development by providing a biologically relevant means of evaluating potential vaccine candidates. Therefore, we aimed to qualify the standard membrane-feeding assay (SMFA. The assay measures the transmission-blocking activity of antibodies by feeding cultured P. falciparum gametocytes to Anopheles mosquitoes in the presence of the test antibodies and measuring subsequent mosquito infection. The International Conference on Harmonisation (ICH Harmonised Tripartite Guideline Q2(R1 details characteristics considered in assay validation. Of these characteristics, we decided to qualify the SMFA for Precision, Linearity, Range and Specificity. The transmission-blocking 4B7 monoclonal antibody was tested over 6 feeding experiments at several concentrations to determine four suitable concentrations that were tested in triplicate in the qualification experiments (3 additional feeds to evaluate Precision, Linearity and Range. For Specificity, 4B7 was tested in the presence of normal mouse IgG. We determined intra- and inter-assay variability of % inhibition of mean oocyst intensity at each concentration of 4B7 (lower concentrations showed higher variability. We also showed that % inhibition was dependent on 4B7 concentration and the activity is specific to 4B7. Since obtaining empirical data is time-consuming, we generated a model using data from all 9 feeds and simulated the effects of different parameters on final readouts to improve the assay procedure and analytical methods for future studies. For example, we estimated the effect of number of mosquitoes dissected on variability of % inhibition, and simulated the relationship between % inhibition in oocyst intensity and % inhibition of prevalence of infected mosquitos at different mean oocysts in the control. SMFA is one of the few biological assays used in

  15. Dose estimation using dicentric chromosome assay and cytokinesis block micronucleus assay: comparison between manual and automated scoring in triage mode.

    Science.gov (United States)

    De Amicis, Andrea; De Sanctis, Stefania; Di Cristofaro, Sara; Franchini, Valeria; Regalbuto, Elisa; Mammana, Giacomo; Lista, Florigio

    2014-06-01

    In cases of an accidental overexposure to ionizing radiation, it is essential to estimate the individual absorbed dose of a potentially radiation-exposed person. For this purpose, biological dosimetry can be performed to confirm, complement or even replace physical dosimetry when this proves to be unavailable. The most validated biodosimetry techniques for dose estimation are the dicentric chromosome assay, the "gold standard" for individual dose assessment, and cytokinesis-block micronucleus assay. However, both assays are time consuming and require skilled scorers. In case of large-scale accidents, different strategies have been developed to increase the throughput of cytogenetic service laboratories. These are the decrease of cell numbers to be scored for triage dosimetry; the automation of procedures including the scoring of, for example, aberrant chromosomes and micronuclei; and the establishment of laboratory networks in order to enable mutual assistance if necessary. In this study, the authors compared the accuracy of triage mode biodosimetry by dicentric chromosome analysis and the cytokinesis block micronucleus assay performing both the manual and the automated scoring mode. For dose estimation using dicentric chromosome assay of 10 blind samples irradiated up to 6.4 Gy of x-rays, a number of metaphase spreads were analyzed ranging from 20 up to 50 cells for the manual and from 20 up to 500 cells for the automatic scoring mode. For dose estimation based on the cytokinesis block micronucleus assay, the micronucleus frequency in both 100 and 200 binucleated cells was determined by manual and automatic scoring. The results of both assays and scoring modes were compared and analyzed considering the sensitivity, specificity, and accuracy of dose estimation with regard to the discrimination power of clinically relevant binary categories of exposure doses.

  16. Critical Appraisal of Information on the Web in Practice: Undergraduate Students’ Knowledge, Reported Use, and Behaviour / Évaluation critique de l’information sur la toile : une vision pratique : les connaissances des étudiants de premier cycle, leur uti

    Directory of Open Access Journals (Sweden)

    Neil R. Hogan

    2012-02-01

    Full Text Available Undergraduates use a wide range of information resources for academic and nonacademic purposes, including web sites that range from credible, peer reviewed, online journal sites, to biased and inaccurate promotional web sites. Students are taught basic critical appraisal skills, but do they apply these skills to make decisions about information in different web sites? In an experimental setting, undergraduate students examined pairs of web sites containing conflicting information based on different aspects of critical appraisal, namely credibility of the author of the information, purpose of the web site, and last update of the site, and answered multiple choice questions about the conflicting information. Results indicated that students failed to use critical appraisal criteria, and that while knowledge of and self-reported use of these criteria were related to each other, they were not related to behaviour. This research demonstrates the need for alternative strategies for critical appraisal instruction and assessment. Les étudiants de premier cycle consultent une vaste gamme de sources d’information à des fins universitaires et non universitaires, y compris des sites Web allant de revues en ligne crédibles et évaluées par des pairs à des sites Web promotionnels partials et inexacts. On enseigne aux étudiants des méthodes de base d’évaluation critique, mais mettent-ils ces méthodes en pratique pour prendre des décisions relativement à l’information tirée de différents sites Web? Dans un cadre expérimental, les étudiants de premier cycle ont étudié des paires de sites Web contenant des informations contradictoires en se fondant sur différents aspects de l’évaluation critique, notamment la crédibilité de l’auteur de l’information, l’intention du site Web et la dernière mise à jour du site, et ont répondu à des questions à choix multiples concernant les informations contradictoires. Les r

  17. Interpreting Skills of Political Terms with Chinese Characteristics Guided by Skopos Theory-a Case Study of Premier Li Keqiang's Press Conference Interpreting%目的论视角下中国特色政治词汇口译策略浅析--以2013年两会期间李克强总理答记者问为例

    Institute of Scientific and Technical Information of China (English)

    周爽; 曾景婷

    2014-01-01

    文章旨在探讨译员是如何运用目的论来口译中国特色政治词汇,实现对外交流的目的。以功能翻译目的论为理论依据,对李克强总理的一场记者招待会现场汉英口译进行了分析。研究结果显示:与其它领域的口译相比,外交口译的特点要求在传译时措辞更严谨,译员会采取不同的口译策略,在目的原则的统领下,遵循目的论的“语内连贯原则”并选择性的违反“语际连贯原则”,即是说为了产出内容能被听众理解,译员需要对口译中目标语作出相应的调整。%This paper aims at analyzing the skills employed to interpret political terms with Chinese Characteristics for foreign ex-change. According to Skopos theory,this paper conducts a case study of a press conference held by Premier Li Keqiang. This re-search demonstrates that even if the nature of diplomatic interpreting calls for a more stringent wording than other kinds of inter-preting,the interpreter always follows the intra-textual coherence rule,adjusting the target text while violating the inter-textual coherence rule to some extent. In other words, in order to realize the purpose of producing an understandable target text, it is necessary to make adjustments to the target text.

  18. The Origin and Development of the Sudanese Ports (‘Aydhâb, Bâ/di‘, Sawâkinin the early Islamic Period Origine et développement des ports soudanais (‘Aydhâb, Bâ/di‘, Sawâkin aux premiers siècles de l’Islam

    Directory of Open Access Journals (Sweden)

    Tim Power

    2008-04-01

    Full Text Available Avant que la mobilité et les échanges liés au /hajj et au commerce avec l’Inde ne s’imposent de manière prédominante dans la mer Rouge médiévale à partir du xie siècle, les ports soudanais connurent une première phase d’activité (632-969, largement ignorée des historiens. Etablis dès les premiers temps de l’Islam, les ports de ‘Aydhâb et de Bâ/di‘ eurent pour première fonction d’appuyer la conquête islamique et de contenir la puissance d’Aksûm en Ethiopie. Les échanges marchands ne se développèrent que lentement. Dans la seconde moitié du viiie siècle se mit en place la traite des esclaves africains, avant que la « ruée vers l’or » du ixe siècle n’accélère la croissance des ports soudanais, entraînant notamment l’établissement du port de Sawâkin.The origin and early development of the Sudanese ports [AD 632-969] has been obscured by a fixation with the medieval ‘India trade’ and /hajj traffic. It can alternatively be shown that ‘Aidhâb and Bâ/di‘ were established to further the Muslim conquests and contain Aksumite Ethiopia. Trade was slow to develop. An African slave trade emerged in the mid eighth century. Gold mining thereafter assumed central importance, with Souakin established in a ninth-century ‘gold rush.’

  19. Application of statistical process control to qualitative molecular diagnostic assays

    LENUS (Irish Health Repository)

    O'Brien, Cathal P.

    2014-11-01

    Modern pathology laboratories and in particular high throughput laboratories such as clinical chemistry have developed a reliable system for statistical process control (SPC). Such a system is absent from the majority of molecular laboratories and where present is confined to quantitative assays. As the inability to apply SPC to an assay is an obvious disadvantage this study aimed to solve this problem by using a frequency estimate coupled with a confidence interval calculation to detect deviations from an expected mutation frequency. The results of this study demonstrate the strengths and weaknesses of this approach and highlight minimum sample number requirements. Notably, assays with low mutation frequencies and detection of small deviations from an expected value require greater sample numbers to mitigate a protracted time to detection. Modeled laboratory data was also used to highlight how this approach might be applied in a routine molecular laboratory. This article is the first to describe the application of SPC to qualitative laboratory data.

  20. Kynetic resazurin assay (KRA) for bacterial quantification of foodborne pathogens

    Science.gov (United States)

    Arenas, Yaxal; Mandel, Arkady; Lilge, Lothar

    2012-03-01

    Fast detection of bacterial concentrations is important for the food industry and for healthcare. Early detection of infections and appropriate treatment is essential since, the delay of treatments for bacterial infections tends to be associated with higher mortality rates. In the food industry and in healthcare, standard procedures require the count of colony-forming units in order to quantify bacterial concentrations, however, this method is time consuming and reports require three days to be completed. An alternative is metabolic-colorimetric assays which provide time efficient in vitro bacterial concentrations. A colorimetric assay based on Resazurin was developed as a time kinetic assay (KRA) suitable for bacterial concentration measurements. An optimization was performed by finding excitation and emission wavelengths for fluorescent acquisition. A comparison of two non-related bacteria, foodborne pathogens Escherichia coli and Listeria monocytogenes, was performed in 96 well plates. A metabolic and clonogenic dependence was established for fluorescent kinetic signals.