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Sample records for amplification reaction pear

  1. Polymerase-endonuclease amplification reaction (PEAR for large-scale enzymatic production of antisense oligonucleotides.

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    Xiaolong Wang

    Full Text Available Antisense oligonucleotides targeting microRNAs or their mRNA targets prove to be powerful tools for molecular biology research and may eventually emerge as new therapeutic agents. Synthetic oligonucleotides are often contaminated with highly homologous failure sequences. Synthesis of a certain oligonucleotide is difficult to scale up because it requires expensive equipment, hazardous chemicals and a tedious purification process. Here we report a novel thermocyclic reaction, polymerase-endonuclease amplification reaction (PEAR, for the amplification of oligonucleotides. A target oligonucleotide and a tandem repeated antisense probe are subjected to repeated cycles of denaturing, annealing, elongation and cleaving, in which thermostable DNA polymerase elongation and strand slipping generate duplex tandem repeats, and thermostable endonuclease (PspGI cleavage releases monomeric duplex oligonucleotides. Each round of PEAR achieves over 100-fold amplification. The product can be used in one more round of PEAR directly, and the process can be further repeated. In addition to avoiding dangerous materials and improved product purity, this reaction is easy to scale up and amenable to full automation. PEAR has the potential to be a useful tool for large-scale production of antisense oligonucleotide drugs.

  2. Comparative genomic analysis reveals multiple long terminal repeats, lineage-specific amplification, and frequent interelement recombination for Cassandra retrotransposon in pear (Pyrus bretschneideri Rehd.).

    Science.gov (United States)

    Yin, Hao; Du, Jianchang; Li, Leiting; Jin, Cong; Fan, Lian; Li, Meng; Wu, Jun; Zhang, Shaoling

    2014-06-04

    Cassandra transposable elements belong to a specific group of terminal-repeat retrotransposons in miniature (TRIM). Although Cassandra TRIM elements have been found in almost all vascular plants, detailed investigations on the nature, abundance, amplification timeframe, and evolution have not been performed in an individual genome. We therefore conducted a comprehensive analysis of Cassandra retrotransposons using the newly sequenced pear genome along with four other Rosaceae species, including apple, peach, mei, and woodland strawberry. Our data reveal several interesting findings for this particular retrotransposon family: 1) A large number of the intact copies contain three, four, or five long terminal repeats (LTRs) (∼20% in pear); 2) intact copies and solo LTRs with or without target site duplications are both common (∼80% vs. 20%) in each genome; 3) the elements exhibit an overall unbiased distribution among the chromosomes; 4) the elements are most successfully amplified in pear (5,032 copies); and 5) the evolutionary relationships of these elements vary among different lineages, species, and evolutionary time. These results indicate that Cassandra retrotransposons contain more complex structures (elements with multiple LTRs) than what we have known previously, and that frequent interelement unequal recombination followed by transposition may play a critical role in shaping and reshaping host genomes. Thus this study provides insights into the property, propensity, and molecular mechanisms governing the formation and amplification of Cassandra retrotransposons, and enhances our understanding of the structural variation, evolutionary history, and transposition process of LTR retrotransposons in plants. © The Author(s) 2014. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  3. An aptasensor for staphylococcus aureus based on nicking enzyme amplification reaction and rolling circle amplification.

    Science.gov (United States)

    Xu, Jingguo; Guo, Jia; Maina, Sarah Wanjiku; Yang, Yumeng; Hu, Yimin; Li, Xuanxuan; Qiu, Jiarong; Xin, Zhihong

    2018-05-15

    An ultra-sensitive aptamer-based biosensor for the detection of staphylococcus aureus was established by adopting the nicking enzyme amplification reaction (NEAR) and the rolling circle amplification (RCA) technologies. Aptamer-probe (AP), containing an aptamer and a probe sequence, was developed to act as the recognition unit of the biosensor, which was specifically bound to S. aureus. The probe was released from AP and initiated into the subsequent DNA amplification reactions where S. aureus was present, converting the detection of S. aureus to the investigation of probe oligonucleotide. The RCA amplification products contained a G-quadruplex motif and formed a three dimensional structure in presence of hemin. The G4/hemin complex showed horseradish peroxidase (HRP)-mimic activity and catalyzed the chemiluminescence reaction of luminol mediated by H 2 O 2 . The results showed that the established biosensor could detect S. aureus specifically with a good linear correlation at 5-10 4  CFU/mL. The signal values based on NEAR-RCA two-step cycle were boosted acutely, much higher than that relied on one-cycle magnification. The limit of detection (LoD) was determined to be as low as 5 CFU/mL. The established aptasensor exhibited a good discrimination of living against dead S. aureus, and can be applied to detect S. aureus in the food industry. Copyright © 2018 Elsevier Inc. All rights reserved.

  4. Mechanism of chimera formation during the Multiple Displacement Amplification reaction

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    Stockwell Timothy B

    2007-04-01

    Full Text Available Abstract Background Multiple Displacement Amplification (MDA is a method used for amplifying limiting DNA sources. The high molecular weight amplified DNA is ideal for DNA library construction. While this has enabled genomic sequencing from one or a few cells of unculturable microorganisms, the process is complicated by the tendency of MDA to generate chimeric DNA rearrangements in the amplified DNA. Determining the source of the DNA rearrangements would be an important step towards reducing or eliminating them. Results Here, we characterize the major types of chimeras formed by carrying out an MDA whole genome amplification from a single E. coli cell and sequencing by the 454 Life Sciences method. Analysis of 475 chimeras revealed the predominant reaction mechanisms that create the DNA rearrangements. The highly branched DNA synthesized in MDA can assume many alternative secondary structures. DNA strands extended on an initial template can be displaced becoming available to prime on a second template creating the chimeras. Evidence supports a model in which branch migration can displace 3'-ends freeing them to prime on the new templates. More than 85% of the resulting DNA rearrangements were inverted sequences with intervening deletions that the model predicts. Intramolecular rearrangements were favored, with displaced 3'-ends reannealing to single stranded 5'-strands contained within the same branched DNA molecule. In over 70% of the chimeric junctions, the 3' termini had initiated priming at complimentary sequences of 2–21 nucleotides (nts in the new templates. Conclusion Formation of chimeras is an important limitation to the MDA method, particularly for whole genome sequencing. Identification of the mechanism for chimera formation provides new insight into the MDA reaction and suggests methods to reduce chimeras. The 454 sequencing approach used here will provide a rapid method to assess the utility of reaction modifications.

  5. Single primer amplification reaction methods reveal exotic and ...

    Indian Academy of Sciences (India)

    Unknown

    mulberry varieties using three different PCR based single primer amplification ..... the results of a multi- variate analysis using Mahalanobis D2 statistic in case of .... Rajan M V, Chaturvedi H K and Sarkar A 1997 Multivariate analysis as an aid ...

  6. Role amplification of the coulomb interaction in nuclear reactions

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Ashok; Soni, S K; Pancholi, S K; Gupta, S L [AN SSSR, Moscow. Radiotekhnicheskij Inst.

    1976-10-01

    The genarally adopted estimate of coulomb interaction in nuclear reactions based on the comparison of relative energies of real particles participating in the reaction with the coulomb barrier has been shown to provide wrong presentation of the role of coulomb interaction in the reaction mechanism. The relative energy of particles participating in virtual processes forming the reaction mechanism and its relation to the coulomb barrier turn out to be tens of per cent less than for the particles in an inlet channel. This is the main reason of increasing the role of coulomb interaction in the reaction mechanism. This increase is particularly significant for nuclei with large charges, in particular, in heavy ion reaction.

  7. Rapid detection of microbial DNA by a novel isothermal genome exponential amplification reaction (GEAR) assay.

    Science.gov (United States)

    Prithiviraj, Jothikumar; Hill, Vincent; Jothikumar, Narayanan

    2012-04-20

    In this study we report the development of a simple target-specific isothermal nucleic acid amplification technique, termed genome exponential amplification reaction (GEAR). Escherichia coli was selected as the microbial target to demonstrate the GEAR technique as a proof of concept. The GEAR technique uses a set of four primers; in the present study these primers targeted 5 regions on the 16S rRNA gene of E. coli. The outer forward and reverse Tab primer sequences are complementary to each other at their 5' end, whereas their 3' end sequences are complementary to their respective target nucleic acid sequences. The GEAR assay was performed at a constant temperature 60 °C and monitored continuously in a real-time PCR instrument in the presence of an intercalating dye (SYTO 9). The GEAR assay enabled amplification of as few as one colony forming units of E. coli per reaction within 30 min. We also evaluated the GEAR assay for rapid identification of bacterial colonies cultured on agar media directly in the reaction without DNA extraction. Cells from E. coli colonies were picked and added directly to GEAR assay mastermix without prior DNA extraction. DNA in the cells could be amplified, yielding positive results within 15 min. Published by Elsevier Inc.

  8. Highly sensitive chemiluminescent point mutation detection by circular strand-displacement amplification reaction.

    Science.gov (United States)

    Shi, Chao; Ge, Yujie; Gu, Hongxi; Ma, Cuiping

    2011-08-15

    Single nucleotide polymorphism (SNP) genotyping is attracting extensive attentions owing to its direct connections with human diseases including cancers. Here, we have developed a highly sensitive chemiluminescence biosensor based on circular strand-displacement amplification and the separation by magnetic beads reducing the background signal for point mutation detection at room temperature. This method took advantage of both the T4 DNA ligase recognizing single-base mismatch with high selectivity and the strand-displacement reaction of polymerase to perform signal amplification. The detection limit of this method was 1.3 × 10(-16)M, which showed better sensitivity than that of most of those reported detection methods of SNP. Additionally, the magnetic beads as carrier of immobility was not only to reduce the background signal, but also may have potential apply in high through-put screening of SNP detection in human genome. Copyright © 2011 Elsevier B.V. All rights reserved.

  9. Probable hypoglycemic adverse drug reaction associated with prickly pear cactus, glipizide, and metformin in a patient with type 2 diabetes mellitus.

    Science.gov (United States)

    Sobieraj, Diana M; Freyer, Craig W

    2010-01-01

    To report a case of an adverse drug reaction (ADR) in a patient with type 2 diabetes mellitus taking prickly pear cactus (PPC), glipizide, and metformin. A 58-year-old Mexican male with type 2 diabetes mellitus being treated with metformin 1000 mg twice daily and extended-release glipizide 10 mg daily was referred to the pharmacist for medication education. He denied taking herbal supplements or experiencing hypoglycemia. Two hemoglobin A(1c) values (6.8% and 6.7%) obtained over the past year demonstrated glycemic control, which was supported by his reported fasting blood glucose readings of 113-132 mg/dL. One month later, the patient reported 4 hypoglycemic events with blood glucose readings of 49-68 mg/dL, which resulted in discontinuation of glipizide. One month later, the patient denied any further hypoglycemia. During medication reconciliation he reported consuming crude PPC pads daily for 2 months for glucose control. Literature suggests that PPC has an effect on lowering blood glucose levels in patients with type 2 diabetes mellitus, although few identified data describe ADRs from combining PPC with other agents used in treating type 2 diabetes mellitus. A literature search of MEDLINE (through December 2009) using the search terms diabetes mellitus, prickly pear cactus, nopal, opuntia, metformin, glipizide, glyburide, glimepiride, and sulfonylurea revealed no case reports of the described ADR. One case report describing the blood glucose-lowering effect of PPC in a patient concurrently taking oral antihyperglycemics documented an episode of hypoglycemia, although the Naranjo probability scale was not applied. One patient survey discovered the most common drug-herbal interaction in the given population to be between PPC and antihyperglycemic agents, resulting in hypoglycemia. In our case, use of the Naranjo probability scale suggests the ADR to be probable. The mechanism may be due to the additive glucose lowering of the 3 agents consumed concurrently by the

  10. Organocatalytic aza-Michael/retro-aza-Michael reaction: pronounced chirality amplification in aza-Michael reaction and racemization via retro-aza-Michael reaction.

    Science.gov (United States)

    Cai, Yong-Feng; Li, Li; Luo, Meng-Xian; Yang, Ke-Fang; Lai, Guo-Qiao; Jiang, Jian-Xiong; Xu, Li-Wen

    2011-05-01

    A detailed experimental investigation of an aza-Michael reaction of aniline and chalcone is presented. A series of Cinchona alkaloid-derived organocatalysts with different functional groups were prepared and used in the aza-Michael and retro-aza-Michael reaction. There was an interesting finding that a complete reversal of stereoselectivity when a benzoyl group was introduced to the cinchonine and cinchonidine. The chirality amplification vs. time proceeds in the quinine-derived organocatalyst containing silicon-based bulky group, QN-TBS, -catalyzed aza-Michael reaction under solvent-free conditions. In addition, we have demonstrated for the first time that racemization was occurred in suitable solvents under mild conditions due to retro-aza-Michael reaction of the Michael adduct of aniline with chalcone. These indicate the equilibrium of retro-aza-Michael reaction and aza-Michael reaction produce the happening of chirality amplification in aza-Michael reaction and racemization via retro-aza-Michael reaction under different conditions, which would be beneficial to the development of novel chiral catalysts for the aza-Michael reactions. Copyright © 2011 Wiley-Liss, Inc.

  11. Monodisperse Picoliter Droplets for Low-Bias and Contamination-Free Reactions in Single-Cell Whole Genome Amplification.

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    Yohei Nishikawa

    Full Text Available Whole genome amplification (WGA is essential for obtaining genome sequences from single bacterial cells because the quantity of template DNA contained in a single cell is very low. Multiple displacement amplification (MDA, using Phi29 DNA polymerase and random primers, is the most widely used method for single-cell WGA. However, single-cell MDA usually results in uneven genome coverage because of amplification bias, background amplification of contaminating DNA, and formation of chimeras by linking of non-contiguous chromosomal regions. Here, we present a novel MDA method, termed droplet MDA, that minimizes amplification bias and amplification of contaminants by using picoliter-sized droplets for compartmentalized WGA reactions. Extracted DNA fragments from a lysed cell in MDA mixture are divided into 105 droplets (67 pL within minutes via flow through simple microfluidic channels. Compartmentalized genome fragments can be individually amplified in these droplets without the risk of encounter with reagent-borne or environmental contaminants. Following quality assessment of WGA products from single Escherichia coli cells, we showed that droplet MDA minimized unexpected amplification and improved the percentage of genome recovery from 59% to 89%. Our results demonstrate that microfluidic-generated droplets show potential as an efficient tool for effective amplification of low-input DNA for single-cell genomics and greatly reduce the cost and labor investment required for determination of nearly complete genome sequences of uncultured bacteria from environmental samples.

  12. Evaluation of the AGCU Expressmarker 16 and 22 PCR Amplification Kits Using Biological Samples Applied to FTA Micro Cards in Reduced Volume Direct PCR Amplification Reactions

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    Samantha J Ogden

    2015-01-01

    Full Text Available This study evaluated the performance of the  Wuxi AGCU ScienTech Incorporation (HuiShan, Wuxi, China AGCU Expressmarker 16 (EX 16 and 22 (EX22 short tandem repeat (STR amplification kits in reduced reaction volumes using direct polymerase chain reaction (PCR amplification workflows. The commercially available PowerPlex® 21 (PP21 System (Promega, Wisconsin, USA, which follows similar direct workflows, was used as a reference. Anticoagulate blood applied to chemically impregnated  FTA TM Micro Cards (GE Healthcare UK Limited, Amersham Place, Little Chalfont, Buckinghamshire, HP7 9NA, UK was used to represent a complex biological sample. Allelic concordance, first-pass success rate, average peak heights, heterozygous peak height ratios (HPHRs, and intracolor and intercolor peak height balance were determined. In reduced volume PCR reactions, the performances of both the EX16 and EX22 STR amplification kits were comparable to that of the PP21 System. The level of performance was maintained at PCR reaction volumes, which are 40% of that recommended. The EX22 and PP21 System kits possess comparable overlapping genome coverage. This study evaluated the performance of the AGCU EX16 and EX22 STR amplification kits in reduced PCR reaction volumes using direct workflows in combination with whole blood applied to FTA TM Micro Cards. Allelic concordance, first-pass success rate, average peak heights, HPHRs, and intracolor and intercolor peak height balance were determined. A concordance analysis was completed that compared the performance of the EX16 and EX22 kits using human blood applied to FTA Micro Cards in combination with full, half, and reduced PCR reaction volumes. The PP21 System (Promega was used as a reference kit. Where appropriate, the distributions of data were assessed using the Shapiro-Wilk test. For normally-distributed data, statistics were calculated using analysis of variance (ANOVA and for nonparametric data the Wilcoxon

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

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    Adrián Ruiz-Villalba

    2017-12-01

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

  14. Hybridization chain reaction amplification for highly sensitive fluorescence detection of DNA with dextran coated microarrays.

    Science.gov (United States)

    Chao, Jie; Li, Zhenhua; Li, Jing; Peng, Hongzhen; Su, Shao; Li, Qian; Zhu, Changfeng; Zuo, Xiaolei; Song, Shiping; Wang, Lianhui; Wang, Lihua

    2016-07-15

    Microarrays of biomolecules hold great promise in the fields of genomics, proteomics, and clinical assays on account of their remarkably parallel and high-throughput assay capability. However, the fluorescence detection used in most conventional DNA microarrays is still limited by sensitivity. In this study, we have demonstrated a novel universal and highly sensitive platform for fluorescent detection of sequence specific DNA at the femtomolar level by combining dextran-coated microarrays with hybridization chain reaction (HCR) signal amplification. Three-dimensional dextran matrix was covalently coated on glass surface as the scaffold to immobilize DNA recognition probes to increase the surface binding capacity and accessibility. DNA nanowire tentacles were formed on the matrix surface for efficient signal amplification by capturing multiple fluorescent molecules in a highly ordered way. By quantifying microscopic fluorescent signals, the synergetic effects of dextran and HCR greatly improved sensitivity of DNA microarrays, with a detection limit of 10fM (1×10(5) molecules). This detection assay could recognize one-base mismatch with fluorescence signals dropped down to ~20%. This cost-effective microarray platform also worked well with samples in serum and thus shows great potential for clinical diagnosis. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. An exonuclease-assisted amplification electrochemical aptasensor for Hg(2+) detection based on hybridization chain reaction.

    Science.gov (United States)

    Bao, Ting; Wen, Wei; Zhang, Xiuhua; Xia, Qinghua; Wang, Shengfu

    2015-08-15

    In this work, a novel electrochemical aptasensor was developed for Hg(2+) detection based on exonuclease-assisted target recycling and hybridization chain reaction (HCR) dual signal amplification strategy. The presence of Hg(2+) induced the T-rich DNA partly folded into duplex-like structure via the Hg(2+) mediated T-Hg(2+)-T base pairs, which triggered the activity of exonuclease III (Exo III). Exo III selectively digested the double-strand DNA containing multiple T-Hg(2+)-T base pairs from its 3'-end, the released Hg(2+) participated analyte recycle. With each digestion cycle, a digestion product named as help DNA was obtained, which acted as a linkage between the capture DNA and auxiliary DNA. The presence of help DNA and two auxiliary DNA collectively facilitated successful HCR process and formed long double-stranded DNA. [Ru(NH3)6](3+) was used as redox indicator, which electrostatically bound to the double strands and produced an electrochemical signal. Exo III-assisted target recycling and HCR dual amplification significantly improved the sensitivity for Hg(2+) with a detection limit of 0.12 pM (S/N=3). Furthermore, the proposed aptasensor had a promising potential for the application of Hg(2+) detection in real aquatic sample analysis. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. DNA amplification fingerprinting using 10 x polymerase chain reaction buffer with ammonium sulfate for human identification

    International Nuclear Information System (INIS)

    Baransel, Asyun; Dugler, Hikmat E.; Tokdemir, Mehmet

    2004-01-01

    The polymerase chain reaction (PCR) - based DNA amplification fingerprinting (DAF) or randomly amplified polymorphic DNA (RAPD) is based on a strategy using a single arbitrary oligonucleotide primer to generate anonymous amplification of genomic DNA. On this basic strategy, in this study, we aimed to test individual differences and usefulness of 2 basic primers (5-CGCGCCGG-3 and 5-TGCCGAGCTG-3) and examined whether there is a positive effect on results of 10 x PCR buffer with ammonium sulfate. A new approach in DNA fingerprinting, 10 x PCR buffer with ammonium sulfate, is presented in the study. Primers with single 8 and 10 nucleotides in length and 2 different PCR buffers with or without ammonium sulfate were used to identify 135 volunteers with no blood relationship. This study was carried out at the Pharmacology Laboratory, University of Gaziantep, School of Medicine, Turkey between 1999 and 2000. An average of 10 major bands representing 500-1500 base pair (bp) in length was determined as amplified DNA products on standard agarose gels for these volunteers. The use of ammonium sulfate in 10 x PCR buffers has increased to 92% success ratio of individual difference obtained from the 8 nucleotides primer. With this study, more reliable results can be obtained by using ammonium sulfate in 10 x PCR buffers. (author)

  17. DNA-based identification of spices: DNA isolation, whole genome amplification, and polymerase chain reaction.

    Science.gov (United States)

    Focke, Felix; Haase, Ilka; Fischer, Markus

    2011-01-26

    Usually spices are identified morphologically using simple methods like magnifying glasses or microscopic instruments. On the other hand, molecular biological methods like the polymerase chain reaction (PCR) enable an accurate and specific detection also in complex matrices. Generally, the origins of spices are plants with diverse genetic backgrounds and relationships. The processing methods used for the production of spices are complex and individual. Consequently, the development of a reliable DNA-based method for spice analysis is a challenging intention. However, once established, this method will be easily adapted to less difficult food matrices. In the current study, several alternative methods for the isolation of DNA from spices have been developed and evaluated in detail with regard to (i) its purity (photometric), (ii) yield (fluorimetric methods), and (iii) its amplifiability (PCR). Whole genome amplification methods were used to preamplify isolates to improve the ratio between amplifiable DNA and inhibiting substances. Specific primer sets were designed, and the PCR conditions were optimized to detect 18 spices selectively. Assays of self-made spice mixtures were performed to proof the applicability of the developed methods.

  18. Detection of Babesia bovis carrier cattle by using polymerase chain reaction amplification of parasite DNA.

    Science.gov (United States)

    Fahrimal, Y; Goff, W L; Jasmer, D P

    1992-01-01

    Carrier cattle infected with Babesia bovis are difficult to detect because of the low numbers of parasites that occur in peripheral blood. However, diagnosis of low-level infections with the parasite is important for evaluating the efficacies of vaccines and in transmission and epidemiological studies. We used the polymerase chain reaction (PCR) to amplify a portion of the apocytochrome b gene from the parasite and tested the ability of this method to detect carrier cattle. The target sequence is associated with a 7.4-kb DNA element in undigested B. bovis genomic DNA (as shown previously), and the amplified product was detected by Southern and dot blot hybridization. The assay was specific for B. bovis, since no amplification was detected with Babesia bigemina, Trypanosoma brucei, Anaplasma marginale, or leukocyte DNA. The target sequence was amplified in DNA from B. bovis Mexico, Texas, and Australia S and L strains, demonstrating the applicability of the method to strains from different geographic regions. The sensitivity of the method ranged from 1 to 10 infected erythrocytes extracted from 0.5 ml of blood. This sensitivity was about 1,000 times greater than that from the use of unamplified parasite DNA. By the PCR method, six B. bovis carrier cattle were detected 86% of the time (range, 66 to 100%) when they were tested 11 times, while with microscopic examination of thick blood smears, the same carrier cattle were detected only 36% of the time (range, 17 to 66%). The method provides a useful diagnostic tool for detecting B. bovis carrier cattle, and the sensitivity is significantly improved over that of current methods. The results also suggest that characteristics of the apocytchrome b gene may make this a valuable target DNA for PCR-based detection of other hemoparasites. Images PMID:1624551

  19. PEAR code review

    International Nuclear Information System (INIS)

    De Wit, R.; Jamieson, T.; Lord, M.; Lafortune, J.F.

    1997-07-01

    As a necessary component in the continuous improvement and refinement of methodologies employed in the nuclear industry, regulatory agencies need to periodically evaluate these processes to improve confidence in results and ensure appropriate levels of safety are being achieved. The independent and objective review of industry-standard computer codes forms an essential part of this program. To this end, this work undertakes an in-depth review of the computer code PEAR (Public Exposures from Accidental Releases), developed by Atomic Energy of Canada Limited (AECL) to assess accidental releases from CANDU reactors. PEAR is based largely on the models contained in the Canadian Standards Association (CSA) N288.2-M91. This report presents the results of a detailed technical review of the PEAR code to identify any variations from the CSA standard and other supporting documentation, verify the source code, assess the quality of numerical models and results, and identify general strengths and weaknesses of the code. The version of the code employed in this review is the one which AECL intends to use for CANDU 9 safety analyses. (author)

  20. PEARS Emission Line Galaxies

    Science.gov (United States)

    Pirzkal, Nor; Rothberg, Barry; Ly, Chun; Rhoads, James E.; Malhotra, Sangeeta; Grogin, Norman A.; Dahlen, Tomas; Meurer, Gerhardt R.; Walsh, Jeremy; Hathi, Nimish P.; hide

    2012-01-01

    We present a full analysis of the Probing Evolution And Reionization Spectroscopically (PEARS) slitless grism spectroscopic data obtained vl'ith the Advanced Camera for Surveys on HST. PEARS covers fields within both the Great Observatories Origins Deep Survey (GOODS) North and South fields, making it ideal as a random surveY of galaxies, as well as the availability of a wide variety of ancillary observations to support the spectroscopic results. Using the PEARS data we are able to identify star forming galaxies within the redshift volume 0 galaxies down to a limiting flux of approx 10 - 18 erg/s/sq cm . The ELRs have also been compared to the properties of the host galaxy, including morphology, luminosity, and mass. From this analysis we find three key results: 1) The computed line luminosities show evidence of a flattening in the luminosity function with increasing redshift; 2) The star forming systems show evidence of disturbed morphologies, with star formation occurring predominantly within one effective (half-light) radius. However, the morphologies show no correlation with host stellar mass; and 3) The number density of star forming galaxies with M(*) >= 10(exp 9) Solar M decreases by an order of magnitude at z<=0.5 relative to the number at 0.5 < z < 0.9 in support of the argument for galaxy downsizing.

  1. Bayesian estimation for quantification by real-time polymerase chain reaction under a branching process model of the DNA molecules amplification process

    NARCIS (Netherlands)

    Lalam, N.; Jacob, C.

    2007-01-01

    The aim of Quantitative Polymerase Chain Reaction is to determine the initial amount X0 of specific nucleic acids from an observed trajectory of the amplification process, the amplification being achieved through successive replication cycles. This process depends on the efficiency fpngn of

  2. Chemical thinning of 'Conference' pears

    NARCIS (Netherlands)

    Maas, F.M.; Kanne, H.J.; Steeg, van der P.A.H.

    2010-01-01

    The increasing difference in the market value of small and larger sized ‘Conference’ pears (>65 mm) and the high labour costs for hand thinning, makes it interesting for growers to find a cheaper and reliable method for thinning pear trees. In 2007, 2008 and 2009 trials were carried out to test

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

    NARCIS (Netherlands)

    Ruiz-Villalba, Adrián; van Pelt-Verkuil, Elizabeth; Gunst, Quinn D.; Ruijter, Jan M.; van den Hoff, Maurice J. B.

    2017-01-01

    Quantitative PCR allows the precise measurement of DNA concentrations and is generally considered to be straightforward and trouble free. However, a survey with 93 validated assays for genes in the Wnt-pathway showed that the amplification of nonspecific products occurs frequently and is unrelated

  4. Resonance amplification of the nuclear reaction X(a,b)Y near the a+X channel threshold

    International Nuclear Information System (INIS)

    Melezhik, V.S.

    1991-01-01

    Deviation of the cross section for the nuclear reaction X(a,b)Y from the Gamow formula due to an interaction additional to the Coulomb one in the entrance channel has been analyzed. It is shown that the reaction cross section has an oscillating structure at low energies. If the maximum of the first oscillation is close to the threshold of the channel a+X, it has a resonance behaviour. The peculiarity of the cross sections leads to the resonance amplification of the rate for a muon-catalyzed fusion reaction ('in flight' fusion) tμ+d→ 4 He+n+μ at the energy =76 eV and may influence the μ-capture rate in a dense mixture of hydrogen isotopes. 26 refs.; 4 figs

  5. Transferability of Newly Developed Pear SSR Markers to Other Rosaceae Species.

    Science.gov (United States)

    Fan, L; Zhang, M-Y; Liu, Q-Z; Li, L-T; Song, Y; Wang, L-F; Zhang, S-L; Wu, J

    2013-01-01

    A set of 120 simple sequence repeats (SSRs) was developed from the newly assembled pear sequence and evaluated for polymorphisms in seven genotypes of pear from different genetic backgrounds. Of these, 67 (55.8 %) primer pairs produced polymorphic amplifications. Together, the 67 SSRs detected 277 alleles with an average of 4.13 per locus. Sequencing of the amplification products from randomly picked loci NAUPy31a and NAUpy53a verified the presence of the SSR loci. When the 67 primer pairs were tested on 96 individual members of eight species in the Rosaceae family, 61.2 % (41/67) of the tested SSRs successfully amplified a PCR product in at least one of the Rosaceae genera. The transferability from pear to different species varied from 58.2 % (apple) to 11.9 % (cherry). The ratio of transferability also reflected the closer relationships within Maloideae over Prunoideae. Two pear SSR markers, NAUpy43c and NAUpy55k, could distinguish the 20 different apple genotypes thoroughly, and UPGMA cluster analysis grouped them into three groups at the similarity level of 0.56. The high level of polymorphism and good transferability of pear SSRs to Rosaceae species indicate their promise for application to future molecular screening, map construction, and comparative genomic studies among pears and other Rosaceae species.

  6. Iodometric microdetermination of phosphorus in organic compounds by use of An amplification reaction

    OpenAIRE

    Al Kubaisi, A. H. [عبد الله حسين الكبيسي; Farag, A. B.; Amin, R. R.

    1994-01-01

    A high chemical amplification method is described for the iodometric microdetermination of phosphorus in organic compounds. This method depends on combustion by the oxygen flask, absorption of the combustion products in distilled water and conversion to orthophosphoric acid with bromine water. The phosphoric acid produced is then allowed to react with sodium molybdate in acid medium to form phosphomolybdic acid which is extracted into a mixture of diethyl ether and pentanol (5:1, V/V), and ba...

  7. 76 FR 54075 - Pears Grown in Oregon and Washington; Assessment Rate Decrease for Fresh Pears

    Science.gov (United States)

    2011-08-31

    ... production research and market development, $6,355,000 for promotion and paid advertising for winter pears, and $1,260,000 for promotion and paid advertising for summer/ fall pears. In comparison, major... advertising for winter pears, and $1,410,000 for promotion and paid advertising for summer/fall pears. The...

  8. Micropropagation of pear (Pyrus sp.).

    Science.gov (United States)

    Reed, Barbara M; Denoma, Jeanine; Wada, Sugae; Postman, Joseph

    2013-01-01

    Elements of micropropagation include establishment of shoot tip cultures, proliferation, rooting, and acclimatization of the resulting plantlets. The wide genetic variation in Pyrus makes micropropagation challenging for many genotypes. Initiation of shoots is most successful from forced dormant shoots or from scions grafted onto seedling rootstocks to impose juvenility. Clean shoots are recovered after testing for contaminants at the initiation stage on ½ strength Murashige and Skoog 1962 medium (MS), at pH 6.9 for 1 week or by streaking on nutrient agar. Although pear species and cultivars are cultured on several well-known media, MS is the most commonly used. Our studies showed that multiplication and growth of shoots are best on Pear Medium with higher concentrations of calcium chloride, potassium phosphate, and magnesium sulfate than MS medium and 4.4 μM N(6) benzyladenine. Pear shoots are often recalcitrant to rooting; however, a 5 s dip in 10 mM indole-3-butyric acid or naphthalene acetic acid before planting on basal medium without plant growth regulators is effective for many genotypes. Pear shoots store well at 1-4°C, and can hold for as long as 4 years without reculture. Cryopreservation protocols are available for long-term storage of pear shoot tips. Acclimation of in vitro-rooted or micrografted shoots in a mist bed follows standard procedures.

  9. Preparation of short cytosine-modified oligonucleotides by nicking enzyme amplification reaction

    Czech Academy of Sciences Publication Activity Database

    Ménová, Petra; Hocek, Michal

    2012-01-01

    Roč. 48, č. 55 (2012), s. 6921-6923 ISSN 1359-7345 R&D Projects: GA ČR GA203/09/0317 Institutional support: RVO:61388963 Keywords : cross - coupling reactions * nucleoside triphosphates * functionalized DNA * restriction endonucleases * polymerase incorporation Subject RIV: CC - Organic Chemistry Impact factor: 6.378, year: 2012

  10. Genomic DNA fingerprinting of clinical Haemophilus influenzae isolates by polymerase chain reaction amplification: comparison with major outer-membrane protein and restriction fragment length polymorphism analysis

    NARCIS (Netherlands)

    van Belkum, A.; Duim, B.; Regelink, A.; Möller, L.; Quint, W.; van Alphen, L.

    1994-01-01

    Non-capsulate strains of Haemophilus influenzae were genotyped by analysis of variable DNA segments obtained by amplification of genomic DNA with the polymerase chain reaction (PCR fingerprinting). Discrete fragments of 100-2000 bp were obtained. The reproducibility of the procedure was assessed by

  11. Scope and Limitations of the Nicking Enzyme Amplification Reaction for the Synthesis of Base-Modified Oligonucleotides and Primers for PCR

    Czech Academy of Sciences Publication Activity Database

    Ménová, Petra; Raindlová, Veronika; Hocek, Michal

    2013-01-01

    Roč. 24, č. 6 (2013), s. 1081-1093 ISSN 1043-1802 R&D Projects: GA ČR GA203/09/0317 Institutional support: RVO:61388963 Keywords : isothermal DNA amplification * cross - coupling reactions * nucleoside triphosphates * polymerase incorporation * functionalized DNA * nucleic-acids Subject RIV: CC - Organic Chemistry Impact factor: 4.821, year: 2013

  12. Easy assessment of diversity in Jatropha curcas L. plants using two single-primer amplification reaction (SPAR) methods

    Energy Technology Data Exchange (ETDEWEB)

    Ranade, Shirish A; Srivastava, Anuj P; Srivastava, Jyoti; Tuli, Rakesh [PMB Division, National Botanical Research Institute, Rana Pratap Marg, Lucknow 226 001, U.P. (India); Rana, Tikam S [Plant Biodiversity and Conservation Biology Division, National Botanical Research Institute, Rana Pratap Marg, Lucknow 226 001, U.P. (India)

    2008-06-15

    Jatropha curcas L. (physic nut) has drawn attention in recent years as a source of seed oil that can provide an economically viable substitute for diesel. Very little work on provenance trials and genetic resources of J. curcas L. has been reported so far. Though J. curcas grows widely in India and several collections of the plant are also maintained, pedigree and provenance records are not always available. This article reports our studies on the diversity amongst the accessions of J. curcas L., both amongst already held collections as well as from a few locations in the wild. Two single-primer amplification reaction (SPAR) methods were used for this purpose. The accessions from the North East were most distant from all other accessions in UPGMA analysis. The NBRI, Bhubaneshwar and Lalkuan accessions were more related to each other. The UPGMA tree clearly shows well-separated accession groups: NBRI, Bhubaneshwar, North East, Lalkuan and Outgroup. The study suggests that this relatively recently introduced plant species shows adequate genetic diversity in India and that the SPAR methods are useful for a rapid assessment of the same. The methods provide important tools for analyzing the diversity within the available collections to shortlist the parental lines for adaptability trials and further improvement of Jatropha plants. (author)

  13. Polymerase spiral reaction (PSR): a novel, visual isothermal amplification method for detection of canine parvovirus 2 genomic DNA.

    Science.gov (United States)

    Gupta, Vikas; Chakravarti, Soumendu; Chander, Vishal; Majumder, Saurabh; Bhat, Shabir Ahmad; Gupta, Vivek Kumar; Nandi, Sukdeb

    2017-07-01

    Canine parvovirus-2 (CPV-2), which is ubiquitously distributed worldwide, causes severe and often fatal gastroenteritis in dogs. Accurate, differential and rapid diagnosis of canine parvoviral enteritis remains a challenge for clinicians. A recently developed isothermal amplification technique, polymerase spiral reaction (PSR), was optimized for the first time for a viral pathogen with reference recombinant plasmid standards from different CPV-2 antigenic variants (CPV-2, CPV-2a, CPV-2b and CPV-2c) and subsequently validated using clinical samples. Addition of chromogenic substrate SYBR Green I after the completion of the reaction resulted in bright green fluorescence in positive samples, while negative samples and a no-template control remained orange. These results were further substantiated through visualization of a laddering pattern of the PSR-amplified product in an agarose gel in positive cases and the absence of this pattern in no-template control and negative samples. The PSR assay was found to be highly specific, as it did not react with other putative canine pathogens (canine adenovirus 1 and canine distemper virus). The sensitivity of the newly developed PSR technique was compared with that of conventional PCR, real-time PCR and LAMP, using a serial tenfold dilution of canine parvovirus DNA. The detection limit of PSR was found to be at the femtogram level, which is comparable with that of real-time PCR and LAMP, which are ten times more sensitive than conventional PCR. The assay was validated using 90 clinical samples, of which 54 were found positive, while only 45 samples were positive in conventional PCR. This novel assay, which is fully compliant with the 'ASSURED' concept for disease diagnosis, provides a simple, rapid, specific, sensitive and cost-effective method for diagnosis of canine parvoviral enteritis in veterinary clinics.

  14. 77 FR 72197 - Pears Grown in Oregon and Washington; Assessment Rate Decrease for Processed Pears

    Science.gov (United States)

    2012-12-05

    ... Agricultural Statistics Service, the total farm-gate value of summer/fall processed pears grown in Oregon and... introductory text and paragraph (a) are revised to read as follows: Sec. 927.237 Processed pear assessment rate...

  15. A cascade autocatalytic strand displacement amplification and hybridization chain reaction event for label-free and ultrasensitive electrochemical nucleic acid biosensing.

    Science.gov (United States)

    Chen, Zhiqiang; Liu, Ying; Xin, Chen; Zhao, Jikuan; Liu, Shufeng

    2018-04-23

    Herein, an autocatalytic strand displacement amplification (ASDA) strategy was proposed for the first time, which was further ingeniously coupled with hybridization chain reaction (HCR) event for the isothermal, label-free and multiple amplification toward nucleic acid detection. During the ASDA module, the target recognition opens the immobilized hairpin probe (IP) and initiates the annealing of the auxiliary DNA strand (AS) with the opened IP for the successive polymerization and nicking reaction in the presence of DNA polymerase and nicking endonuclease. This induces the target recycling and generation of a large amount of intermediate DNA sequences, which can be used as target analogy to execute the autocatalytic strand displacement amplification. Simultaneously, the introduced AS strand can propagate the HCR between two hairpins (H1 and H2) to form a linear DNA concatamer with cytosine (C)-rich loop region, which can facilitate the in-situ synthesis of silver nanoclusters (AgNCs) as electrochemical tags for further amplification toward target responses. With current cascade ASDA and HCR strategy, the detection of target DNA could be achieved with a low detection limit of about 0.16 fM and a good selectivity. The developed biosensor also exhibits the distinct advantages of flexibility and simplicity in probe design and biosensor fabrication, and label-free electrochemical detection, thus opens a promising avenue for the detection of nucleic acid with low abundance in bioanalysis and clinical biomedicine. Copyright © 2018 Elsevier B.V. All rights reserved.

  16. Time Course of Detection of Human Male DNA from Stained Blood Sample on Various Surfaces by Loop Mediated Isothermal Amplification and Polymerase Chain Reaction

    OpenAIRE

    Panan Kanchanaphum

    2018-01-01

    This study explores determining the sex of humans from blood stains taken from different surfaces and compares the time course of detection with the conventional PCR, Conventional Loop Mediated Isothermal Amplification (LAMP), and LAMP-Lateral Flow Dipstick (LFD). For the DNA templates, 7 male and 7 female blood stained samples were extracted and added to LAMP and PCR reaction solution to amplify the SRY gene. The DNA samples were extracted from the following blood stained materials: cloth, w...

  17. Ultrasensitive Faraday cage-type electrochemiluminescence assay for femtomolar miRNA-141 via graphene oxide and hybridization chain reaction-assisted cascade amplification.

    Science.gov (United States)

    Lu, Jing; Wu, Lin; Hu, Yufang; Wang, Sui; Guo, Zhiyong

    2018-06-30

    In this study, a novel electrochemiluminescence (ECL) biosensor for sensitive detection of femtomolar miRNA-141 was constructed on the basis of Faraday cage-type strategy via graphene oxide (GO) and hybridization chain reaction (HCR)-assisted cascade amplification. A capture probe (CP) was immobilized on Fe 3 O 4 @SiO 2 @Au nanoparticles as capture unit, which could catch the miRNA-141, and the immobilization of the signal unit (Ru(phen) 3 2+ -HCR/GO) was allowed via nucleic acid hybridization. The prepared biosensor exhibited two advantages for signal amplification: firstly, GO could lap on the electrode surface directly, extending Outer Helmholtz Plane (OHP) of the sensor due to the large surface area and good electronic transport property; secondly, HCR-assisted cascade amplification was designed by anchoring all HCR products on the GO surface, then embedding Ru(phen) 3 2+ as a signal readout pathway. All these signal molecules could take part in electrochemical reactions, thus further enhancing the ECL signal drastically. Therefore, the proposed sensor constructed by integrating HCR with Faraday cage-type strategy displayed an ultrasensitive detection platform for the miRNA-141 with a low detection limit of 0.03 fM. In addition, this proposed biosensor provides a universal platform for analysis of other microRNAs. Copyright © 2018 Elsevier B.V. All rights reserved.

  18. Loop mediated isothermal amplification assay using hydroxy naphthol blue, conventional polymerase chain reaction and real-time PCR in the diagnosis of intraocular tuberculosis

    Directory of Open Access Journals (Sweden)

    P K Balne

    2015-01-01

    Full Text Available This study is a comparative evaluation (Chi-square test of a closed tube loop mediated isothermal amplification assay using hydroxy naphthol blue dye (HNB-LAMP, real-time polymerase chain reaction (PCR and conventional PCR in the diagnosis of intraocular tuberculosis. Considering clinical presentation as the gold standard in 33 patients, the sensitivity of HNB-LAMP assay (75.8% was higher (not significant, P value 0.2 than conventional PCR (57.6% and lower than real-time PCR (90.9%. Specificity was 100% by all three methods. No amplification was observed in negative controls (n = 20 by all three methods. The cost of the HNB-LAMP assay was Rs. 500.00 and it does not require thermocycler, therefore, it can be used as an alternative to conventional PCR in resource-poor settings.

  19. Label-free and sensitive detection of T4 polynucleotide kinase activity via coupling DNA strand displacement reaction with enzymatic-aided amplification.

    Science.gov (United States)

    Cheng, Rui; Tao, Mangjuan; Shi, Zhilu; Zhang, Xiafei; Jin, Yan; Li, Baoxin

    2015-11-15

    Several fluorescence signal amplification strategies have been developed for sensitive detection of T4 polynucleotide kinase (T4 PNK) activity, but they need fluorescence dye labeled DNA probe. We have addressed the limitation and report here a label-free strategy for sensitive detection of PNK activity by coupling DNA strand displacement reaction with enzymatic-aided amplification. A hairpin oligonucleotide (hpDNA) with blunt ends was used as the substrate for T4 PNK phosphorylation. In the presence of T4 PNK, the stem of hpDNA was phosphorylated and further degraded by lambda exonuclease (λ exo) from 5' to 3' direction to release a single-stranded DNA as a trigger of DNA strand displacement reaction (SDR). The trigger DNA can continuously displace DNA P2 from P1/P2 hybrid with the help of specific cleavage of nicking endonuclease (Nt.BbvCI). Then, DNA P2 can form G-quadruplex in the presence of potassium ions and quadruplex-selective fluorphore, N-methyl mesoporphyrin IX (NMM), resulting in a significant increase in fluorescence intensity of NMM. Thus, the accumulative release of DNA P2 led to fluorescence signal amplification for determining T4 PNK activity with a detection limit of 6.6×10(-4) U/mL, which is superior or comparative with established approaches. By ingeniously utilizing T4 PNK-triggered DNA SDR, T4 PNK activity can be specifically and facilely studied in homogeneous solution containing complex matrix without any external fluorescence labeling. Moreover, the influence of different inhibitors on the T4 PNK activity revealed that it also can be explored to screen T4 PNK inhibitors. Therefore, this label-free amplification strategy presents a facile and cost-effective approach for nucleic acid phosphorylation related research. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Silver nanoclusters-assisted ion-exchange reaction with CdTe quantum dots for photoelectrochemical detection of adenosine by target-triggering multiple-cycle amplification strategy.

    Science.gov (United States)

    Zhao, Yang; Tan, Lu; Gao, Xiaoshan; Jie, Guifen; Huang, Tingyu

    2018-07-01

    Herein, we successfully devised a novel photoelectrochemical (PEC) platform for ultrasensitive detection of adenosine by target-triggering cascade multiple cycle amplification based on the silver nanoparticles-assisted ion-exchange reaction with CdTe quantum dots (QDs). In the presence of target adenosine, DNA s1 is released from the aptamer and then hybridizes with hairpin DNA (HP1), which could initiate the cycling cleavage process under the reaction of nicking endonuclease. Then the product (DNA b) of cycle I could act as the "DNA trigger" of cycle II to further generate a large number of DNA s1, which again go back to cycle I, thus a cascade multiple DNA cycle amplification was carried out to produce abundant DNA c. These DNA c fragments with the cytosine (C)-rich loop were captured by magnetic beads, and numerous silver nanoclusters (Ag NCs) were synthesized by AgNO 3 and sodium borohydride. The dissolved AgNCs released numerous silver ions which could induce ion exchange reaction with the CdTe QDs, thus resulting in greatly amplified change of photocurrent for target detection. The detection linear range for adenosine was 1.0 fM ~10 nM with the detection limit of 0.5 fM. The present PEC strategy combining cascade multiple DNA cycle amplification and AgNCs-induced ion-exchange reaction with QDs provides new insight into rapid, and ultrasensitive PEC detection of different biomolecules, which showed great potential for detecting trace amounts in bioanalysis and clinical biomedicine. Copyright © 2018 Elsevier B.V. All rights reserved.

  1. A note on the Noyori model for chiral amplification in the aminoalcohol-catalyzed reaction of aldehydes with dialkylzinc

    Directory of Open Access Journals (Sweden)

    IVAN GUTMAN

    1999-11-01

    Full Text Available The Noyori model of chiral amplification in the alkylation of aldehydes by means of dialkylzinc, catalyzed by chiral aminoalcohols, is further elaborated. A direct, but approximate, relation is obtained between the enantiomeric excess of the catalyst added and the enantiomeric excess of the product.

  2. Toehold-mediated strand displacement reaction triggered isothermal DNA amplification for highly sensitive and selective fluorescent detection of single-base mutation.

    Science.gov (United States)

    Zhu, Jing; Ding, Yongshun; Liu, Xingti; Wang, Lei; Jiang, Wei

    2014-09-15

    Highly sensitive and selective detection strategy for single-base mutations is essential for risk assessment of malignancy and disease prognosis. In this work, a fluorescent detection method for single-base mutation was proposed based on high selectivity of toehold-mediated strand displacement reaction (TSDR) and powerful signal amplification capability of isothermal DNA amplification. A discrimination probe was specially designed with a stem-loop structure and an overhanging toehold domain. Hybridization between the toehold domain and the perfect matched target initiated the TSDR along with the unfolding of the discrimination probe. Subsequently, the target sequence acted as a primer to initiate the polymerization and nicking reactions, which released a great abundant of short sequences. Finally, the released strands were annealed with the reporter probe, launching another polymerization and nicking reaction to produce lots of G-quadruplex DNA, which could bind the N-methyl mesoporphyrin IX to yield an enhanced fluorescence response. However, when there was even a single base mismatch in the target DNA, the TSDR was suppressed and so subsequent isothermal DNA amplification and fluorescence response process could not occur. The proposed approach has been successfully implemented for the identification of the single-base mutant sequences in the human KRAS gene with a detection limit of 1.8 pM. Furthermore, a recovery of 90% was obtained when detecting the target sequence in spiked HeLa cells lysate, demonstrating the feasibility of this detection strategy for single-base mutations in biological samples. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. A Complementary Isothermal Amplification Method to the U.S. EPA Quantitative Polymerase Chain Reaction Approach for the Detection of Enterococci in Environmental Waters.

    Science.gov (United States)

    Kolm, Claudia; Martzy, Roland; Brunner, Kurt; Mach, Robert L; Krska, Rudolf; Heinze, Georg; Sommer, Regina; Reischer, Georg H; Farnleitner, Andreas H

    2017-06-20

    We report a novel molecular assay, based on helicase-dependent amplification (HDA), for the detection of enterococci as markers for fecal pollution in water. This isothermal assay targets the same Enterococcus 23S rRNA gene region as the existing quantitative polymerase chain reaction (qPCR) assays of U.S. Environmental Protection Agency Methods 1611 and 1609 but can be entirely performed on a simple heating block. The developed Enterococcus HDA assay successfully discriminated 15 enterococcal from 15 non-enterococcal reference strains and reliably detected 48 environmental isolates of enterococci. The limit of detection was 25 target copies per reaction, only 3 times higher than that of qPCR. The applicability of the assay was tested on 30 environmental water sample DNA extracts, simulating a gradient of fecal pollution. Despite the isothermal nature of the reaction, the HDA results were consistent with those of the qPCR reference. Given this performance, we conclude that the developed Enterococcus HDA assay has great potential as a qualitative molecular screening method for resource-limited settings when combined with compatible up- and downstream processes. This amplification strategy can pave the way for developing a new generation of rapid, low-cost, and field-deployable molecular diagnostic tools for water quality monitoring.

  4. Invading stacking primer: A trigger for high-efficiency isothermal amplification reaction with superior selectivity for detecting microRNA variants.

    Science.gov (United States)

    Liu, Weipeng; Zhu, Minjun; Liu, Hongxing; Wei, Jitao; Zhou, Xiaoming; Xing, Da

    2016-07-15

    Searching for a strategy to enhance the efficiency of nucleic acid amplification and achieve exquisite discrimination of nucleic acids at the single-base level for biological detection has become an exciting research direction in recent years. Here, we have developed a simple and universal primer design strategy which produces a fascinating effect on isothermal strand displacement amplification (iSDA). We refer to the resultant primer as "invading stacking primer (IS-Primer)" which is based on contiguous stacking hybridization and toehold-mediated exchange reaction and function by merely changing the hybridization location of the primer. Using the IS-Primer, the sensitivity in detecting the target miR-21 is improved approximately five fold compared with the traditional iSDA reaction. It was further demonstrated that the IS-Primer acts as an invading strand to initiate branch migration which can increase the efficiency of the untwisting of the hairpin probe. This effect is equivalent to reducing the free energy of the stem, and the technique shows superior selectivity for single-base mismatches. By demonstrating the enhanced effect of the IS-Primer in the iSDA reaction, this work may provide a potentially new avenue for developing more sensitive and selective nucleic acids assays. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Ultrasensitive electrochemical detection of DNA based on Zn²⁺ assistant DNA recycling followed with hybridization chain reaction dual amplification.

    Science.gov (United States)

    Qian, Yong; Wang, Chunyan; Gao, Fenglei

    2015-01-15

    A new strategy to combine Zn(2+) assistant DNA recycling followed with hybridization chain reaction dual amplification was designed for highly sensitive electrochemical detection of target DNA. A gold electrode was used to immobilize molecular beacon (MB) as the recognition probe and perform the amplification procedure. In the presence of the target DNA, the hairpin probe 1 was opened, and the DNAzyme was liberated from the caged structure. The activated DNAzyme hybridized with the MB and catalyzed its cleavage in the presence of Zn(2+) cofactor and resulting in a free DNAzyme strand. Finally, each target-induced activated DNAzyme underwent many cycles triggering the cleavage of MB, thus forming numerous MB fragments. The MB fragments triggered the HCR and formed a long double-helix DNA structure. Because both H1 and H2 were labeled by biotin, a lot of SA-ALP was captured on the electrode surface, thus catalyzing a silver deposition process for electrochemical stripping analysis. This novel cascade signal amplification strategy can detect target DNA down to the attomolar level with a dynamic range spanning 6 orders of magnitude. This highly sensitive and specific assay has a great potential to become a promising DNA quantification method in biomedical research and clinical diagnosis. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. 21 CFR 145.175 - Canned pears.

    Science.gov (United States)

    2010-04-01

    ... CONSUMPTION CANNED FRUITS Requirements for Specific Standardized Canned Fruits § 145.175 Canned pears. (a.... The lower end of the rod is a plane surface to which the vertical axis of the rod is perpendicular...

  7. Identification of ochratoxin A producing Aspergillus carbonarius and A. niger clade isolated from grapes using the loop-mediated isothermal amplification (LAMP) reaction.

    Science.gov (United States)

    Storari, M; von Rohr, R; Pertot, I; Gessler, C; Broggini, G A L

    2013-04-01

    To develop two assays based on the loop-mediated isothermal amplification (LAMP) of DNA for the quick and specific identification of Aspergillus carbonarius and ochratoxigenic strains of the Aspergillus niger clade isolated from grapes. Two sets of primers were designed based on the polyketide synthase genes involved or putatively involved in ochratoxin A (OTA) biosynthesis in A. carbonarius and A. niger clade. Hydroxynaphthol blue was used as indirect method to indicate DNA amplification. The limit of detection of both assays was comparable to that of a PCR reaction. Specificities of the reactions were tested using DNA from different black aspergilli isolated from grapes. The two LAMP assays were then used to identify A. carbonarius and ochratoxigenic A. niger and A. awamori grown in pure cultures without a prior DNA extraction. The two LAMP assays permitted to quickly and specifically identify DNA from OTA-producing black aspergilli, as well as isolates grown in pure culture. Monitoring vineyards for the presence of OTA-producing strains is part of the measures to minimize the occurrence of OTA in grape products. The two LAMP assays developed here could be potentially used to speed the screening process of vineyards for the presence of OTA-producing black aspergilli. © 2013 The Society for Applied Microbiology.

  8. Direct RNA detection without nucleic acid purification and PCR: Combining sandwich hybridization with signal amplification based on branched hybridization chain reaction.

    Science.gov (United States)

    Xu, Yao; Zheng, Zhi

    2016-05-15

    We have developed a convenient, robust and low-cost RNA detection system suitable for high-throughput applications. This system uses a highly specific sandwich hybridization to capture target RNA directly onto solid support, followed by on-site signal amplification via 2-dimensional, branched hybridizing chain polymerization through toehold-mediated strand displacement reaction. The assay uses SYBR Green to detect targets at concentrations as low as 1 pM, without involving nucleic acid purification or any enzymatic reaction, using ordinary oligonucleotides without modification or labeling. The system was demonstrated in the detection of malaria RNA in blood and GAPDH gene expression in cell lysate. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Droplet digital polymerase chain reaction detection of HER2 amplification in formalin fixed paraffin embedded breast and gastric carcinoma samples.

    Science.gov (United States)

    Zhu, Yazhen; Lu, Dan; Lira, Maruja E; Xu, Qing; Du, Yunzhi; Xiong, Jianghong; Mao, Mao; Chung, Hyun Cheol; Zheng, Guangjuan

    2016-04-01

    Human epidermal growth factor receptor 2 (HER2) is a key driver of tumorigenesis, and over-expression as a result of HER2 gene amplification has been observed in a number of solid tumors. Recently HER2 has become an important biomarker for the monoclonal antibody treatment of HER2-positive metastatic breast and advanced gastric cancer. The HER2 targeting antibody trastuzumab treatment requires accurate measurement of HER2 levels for proper diagnosis. Droplet digital PCR (ddPCR) with highly direct, precise and absolute nucleic acid quantification could be used to detect HER2 amplification levels. Our objective was to evaluate a robust, accurate and less subjective application of ddPCR for HER2 amplification levels and test the assay performance in clinical formalin-fixed paraffin-embedded (FFPE) breast and gastric carcinoma samples. Genomic DNA from HER2 amplified cell line SK-BR-3 was used to set up the ddPCR assays. The copy number of HER2 was compared to the chromosome 17 centromere reference gene (CEP17), expressed as HER2:CEP17 ratio. Genomic DNAs of FFPE specimens from 145 Asian patients with breast and gastric carcinomas were assayed using both standard methods, immunohistochemistry (IHC) and/or fluorescence in situ hybridization (FISH), and ddPCR. Based on 145 clinical breast and gastric carcinoma cases, our study demonstrated a high concordance of ddPCR results to FISH and IHC. In breast cancer specimens, the ddPCR results had high concordance with FISH and IHC defined HER2 status with a sensitivity of 90.9% (30/33) and a specificity of 100% (77/77). In gastric cancer specimens that were concordant in both FISH and IHC, our assay was 95.5% concordant with FISH and IHC (21/22). ddPCR has the advantage of automation and also allows levels of HER2 amplification to be easily evaluated in large numbers of samples, and presents a potential option to define HER2 status. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Time Course of Detection of Human Male DNA from Stained Blood Sample on Various Surfaces by Loop Mediated Isothermal Amplification and Polymerase Chain Reaction

    Directory of Open Access Journals (Sweden)

    Panan Kanchanaphum

    2018-01-01

    Full Text Available This study explores determining the sex of humans from blood stains taken from different surfaces and compares the time course of detection with the conventional PCR, Conventional Loop Mediated Isothermal Amplification (LAMP, and LAMP-Lateral Flow Dipstick (LFD. For the DNA templates, 7 male and 7 female blood stained samples were extracted and added to LAMP and PCR reaction solution to amplify the SRY gene. The DNA samples were extracted from the following blood stained materials: cloth, wood, clay, and tile. Then, the samples were stored at room temperature for 1, 7, 30, and 60 day(s. After the DNA amplification, the gel electrophoresis process was applied to detect LAMP product. The LFD was combined with the LAMP to detect LAMP product on the male cloth samples. For the male samples, the time course of detection on the first and seventh days indicated positive for both LAMP and PCR products on all the surfaces while no DNA amplification was found on any of the female samples. On day 30, positive LAMP product was still found on all the male samples. However, it had faded on the tiles. Moreover, all the male samples, which had tested positive for PCR product, were blurred and unclear. On day 60, LAMP product was still found on all the male samples. Conversely, the PCR method resulted in no bands showing for any of the male samples. However, the LAMP-LFD method detected product on all the male samples of cloth. The results show that the LAMP is an effective, practical, and reliable molecular-biological method. Moreover, the LFD can increase the efficiency and sensitivity of the LAMP, making it more suitable for field studies because gel electrophoresis apparatus is not required.

  11. Molecular detection of field isolates of Turkey Eimeria by polymerase chain reaction amplification of the cytochrome c oxidase I gene.

    Science.gov (United States)

    Rathinam, T; Gadde, U; Chapman, H D

    2015-07-01

    Oocysts of Eimeria spp. were isolated from litter samples obtained from 30 commercial turkey farms. Genomic DNA was extracted from clean oocysts, and polymerase chain amplification of the species-specific cytochrome c oxidase subunit I (COI) gene was performed for five species of turkey Eimeria. The species tested were Eimeria adenoeides, Eimeria meleagrimitis, Eimeria meleagridis, Eimeria dispersa, and Eimeria gallopavonis. All DNA samples were positive for E. meleagrimitis, nine were positive for E. adenoeides, two were positive for E. dispersa, and none for E. meleagridis and E. gallopavonis. E. meleagrimitis occurred as a single species in 21 (70 %) of the farms while 9 (30 %) farms had a mixed species with E. meleagrimitis and E. adenoeides and 2 (7 %) were triple positive with E. meleagrimitis, E. adenoeides, and E. dispersa. This is the first account of the field prevalence of turkey Eimeria species using molecular methods.

  12. 7 CFR 927.103 - Organically produced pears.

    Science.gov (United States)

    2010-01-01

    ... Agreements and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE PEARS GROWN IN OREGON AND... means pears that have been certified by an organic certification organization currently registered with... under the National Organic Program. Communications ...

  13. Polymerase chain reaction amplification and cloning of immunogenic protein NAD-dependent beta hydroxybutyryl CoA dehydrogenase gene of Clostridium chauvoei

    Directory of Open Access Journals (Sweden)

    Saroj K. Dangi

    2014-10-01

    Full Text Available Aim: The present study was aimed at polymerase chain reaction (PCR amplification and cloning of NAD-dependent betahydroxybutyryl coenzyme A dehydrogenase (BHBD gene of Clostridium chauvoei. Materials and Methods: C. chauvoei was cultured and confirmed by 16-23S rDNA spacer region primers. The primers for nad-bhbd gene of C. chauvoei were designed to aid in cloning into pRham-N-His SUMO-Kan vector, and nad-bhbd gene was amplified by PCR. The amplified nad-bhbd gene was purified and cloned into pRham-N-His SUMO-Kan expression vector. The recombinant plasmid was transformed into E. cloni 10 G cells and the clone was confirmed by colony PCR using the pRham-SUMO-NAD-For and pRham-SUMO-NAD-Rev primers and also by sequencing. Results: PCR amplification of nad-bhbd gene yielded a product length of 844 base pairs which was cloned into pRham-NHis SUMO-Kan vector followed by transformation into E. cloni 10G chemically competent cells. The recombinant clones were characterized by colony PCR, sequencing, followed by basic local alignment search tool (BLAST analysis to confirm the insert. Conclusions: Immunogenic protein NAD- dependent BHBD of C. chauvoei was cloned and the recombinant clones were confirmed by colony PCR and sequencing analysis.

  14. Ultrasensitive electrochemical detection of avian influenza A (H7N9) virus DNA based on isothermal exponential amplification coupled with hybridization chain reaction of DNAzyme nanowires.

    Science.gov (United States)

    Yu, Yanyan; Chen, Zuanguang; Jian, Wensi; Sun, Duanping; Zhang, Beibei; Li, Xinchun; Yao, Meicun

    2015-02-15

    In this work, a simple and label-free electrochemical biosensor with duel amplification strategy was developed for DNA detection based on isothermal exponential amplification (EXPAR) coupled with hybridization chain reaction (HCR) of DNAzymes nanowires. Through rational design, neither the primer nor the DNAzymes containing molecular beacons (MBs) could react with the duplex probe which were fixed on the electrode surface. Once challenged with target, the duplex probe cleaved and triggered the EXPAR mediated target recycle and regeneration circles as well as the HCR process. As a result, a greater amount of targets were generated to cleave the duplex probes. Subsequently, the nanowires consisting of the G-quadruplex units were self-assembled through hybridization with the strand fixed on the electrode surface. In the presence of hemin, the resulting catalytic G-quadruplex-hemin HRP-mimicking DNAzymes were formed. Electrochemical signals can be obtained by measuring the increase in reduction current of oxidized 3.3',5.5'-tetramethylbenzidine sulfate (TMB), which was generated by DNAzyme in the presence of H2O2. This method exhibited ultrahigh sensitivity towards avian influenza A (H7N9) virus DNA sequence with detection limits of 9.4 fM and a detection range of 4 orders of magnitude. The biosensor was also capable of discriminating single-nucleotide difference among concomitant DNA sequences and performed well in spiked cell lysates. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Identification of planorbids from Venezuela by polymerase chain reaction amplification and restriction fragment length polymorphism of internal transcriber spacer of the RNA ribosomal gene

    Directory of Open Access Journals (Sweden)

    Caldeira Roberta L

    2000-01-01

    Full Text Available Snails of the genus Biomphalaria from Venezuela were subjected to morphological assessment as well as polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP analysis. Morphological identification was carried out by comparison of characters of the shell and the male and female reproductive apparatus. The PCR-RFLP involved amplification of the internal spacer region ITS1 and ITS2 of the RNA ribosomal gene and subsequent digestion of this fragment by the restriction enzymes DdeI, MnlI, HaeIII and MspI. The planorbids were compared with snails of the same species and others reported from Venezuela and present in Brazil, Cuba and Mexico. All the enzymes showed a specific profile for each species, that of DdeI being the clearest. The snails were identified as B. glabrata, B. prona and B. kuhniana.

  16. 7 CFR 319.56-39 - Fragrant pears from China.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 5 2010-01-01 2010-01-01 false Fragrant pears from China. 319.56-39 Section 319.56-39... pears from China. Fragrant pears may be imported into the United States from China only under the... China. (2) All propagative material introduced into a registered production site must be certified free...

  17. 7 CFR 927.121 - Pears for gift purposes.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false Pears for gift purposes. 927.121 Section 927.121... WASHINGTON Rules and Regulations Exemptions and Safeguards § 927.121 Pears for gift purposes. There are exempted from the provisions of the order any and all pears which, in individual gift packages, are shipped...

  18. Evaluation of Amplification Targets for the Specific Detection of Bordetella pertussis Using Real-Time Polymerase Chain Reaction

    Directory of Open Access Journals (Sweden)

    Mohammad Rubayet Hasan

    2014-01-01

    Full Text Available BACKGROUND: Bordetella pertussis infections continue to be a major public health challenge in Canada. Polymerase chain reaction (PCR assays to detect B pertussis are typically based on the multicopy insertion sequence IS481, which offers high sensitivity but lacks species specificity.

  19. Multiplex Amplification Refractory Mutation System Polymerase Chain Reaction (ARMS-PCR for diagnosis of natural infection with canine distemper virus

    Directory of Open Access Journals (Sweden)

    Wong Min-Liang

    2010-06-01

    Full Text Available Abstract Background Canine distemper virus (CDV is present worldwide and produces a lethal systemic infection of wild and domestic Canidae. Pre-existing antibodies acquired from vaccination or previous CDV infection might interfere the interpretation of a serologic diagnosis method. In addition, due to the high similarity of nucleic acid sequences between wild-type CDV and the new vaccine strain, current PCR derived methods cannot be applied for the definite confirmation of CD infection. Hence, it is worthy of developing a simple and rapid nucleotide-based assay for differentiation of wild-type CDV which is a cause of disease from attenuated CDVs after vaccination. High frequency variations have been found in the region spanning from the 3'-untranslated region (UTR of the matrix (M gene to the fusion (F gene (designated M-F UTR in a few CDV strains. To establish a differential diagnosis assay, an amplification refractory mutation analysis was established based on the highly variable region on M-F UTR and F regions. Results Sequences of frequent polymorphisms were found scattered throughout the M-F UTR region; the identity of nucleic acid between local strains and vaccine strains ranged from 82.5% to 93.8%. A track of AAA residue located 35 nucleotides downstream from F gene start codon highly conserved in three vaccine strains were replaced with TGC in the local strains; that severed as target sequences for deign of discrimination primers. The method established in the present study successfully differentiated seven Taiwanese CDV field isolates, all belonging to the Asia-1 lineage, from vaccine strains. Conclusions The method described herein would be useful for several clinical applications, such as confirmation of nature CDV infection, evaluation of vaccination status and verification of the circulating viral genotypes.

  20. Multiplex Amplification Refractory Mutation System Polymerase Chain Reaction (ARMS-PCR) for diagnosis of natural infection with canine distemper virus.

    Science.gov (United States)

    Chulakasian, Songkhla; Lee, Min-Shiuh; Wang, Chi-Young; Chiou, Shyan-Song; Lin, Kuan-Hsun; Lin, Fong-Yuan; Hsu, Tien-Huan; Wong, Min-Liang; Chang, Tien-Jye; Hsu, Wei-Li

    2010-06-10

    Canine distemper virus (CDV) is present worldwide and produces a lethal systemic infection of wild and domestic Canidae. Pre-existing antibodies acquired from vaccination or previous CDV infection might interfere the interpretation of a serologic diagnosis method. In addition, due to the high similarity of nucleic acid sequences between wild-type CDV and the new vaccine strain, current PCR derived methods cannot be applied for the definite confirmation of CD infection. Hence, it is worthy of developing a simple and rapid nucleotide-based assay for differentiation of wild-type CDV which is a cause of disease from attenuated CDVs after vaccination. High frequency variations have been found in the region spanning from the 3'-untranslated region (UTR) of the matrix (M) gene to the fusion (F) gene (designated M-F UTR) in a few CDV strains. To establish a differential diagnosis assay, an amplification refractory mutation analysis was established based on the highly variable region on M-F UTR and F regions. Sequences of frequent polymorphisms were found scattered throughout the M-F UTR region; the identity of nucleic acid between local strains and vaccine strains ranged from 82.5% to 93.8%. A track of AAA residue located 35 nucleotides downstream from F gene start codon highly conserved in three vaccine strains were replaced with TGC in the local strains; that severed as target sequences for deign of discrimination primers. The method established in the present study successfully differentiated seven Taiwanese CDV field isolates, all belonging to the Asia-1 lineage, from vaccine strains. The method described herein would be useful for several clinical applications, such as confirmation of nature CDV infection, evaluation of vaccination status and verification of the circulating viral genotypes.

  1. GENOMIC DNA-FINGERPRINTING OF CLINICAL HAEMOPHILUS-INFLUENZAE ISOLATES BY POLYMERASE CHAIN-REACTION AMPLIFICATION - COMPARISON WITH MAJOR OUTER-MEMBRANE PROTEIN AND RESTRICTION-FRAGMENT-LENGTH-POLYMORPHISM ANALYSIS

    NARCIS (Netherlands)

    VANBELKUM, A; DUIM, B; REGELINK, A; MOLLER, L; QUINT, W; VANALPHEN, L

    Non-capsulate strains of Haemophilus influenzae were genotyped by analysis of variable DNA segments obtained by amplification of genomic DNA with the polymerase chain reaction (PCR fingerprinting). Discrete fragments of 100-2000 bp were obtained. The reproducibility of the procedure was assessed by

  2. Detection of Brazilian hantavirus by reverse transcription polymerase chain reaction amplification of N gene in patients with hantavirus cardiopulmonary syndrome

    OpenAIRE

    Marcos Lázaro Moreli; Ricardo Luiz Moro de Sousa; Luiz Tadeu Moraes Figueiredo

    2004-01-01

    We report a nested reverse transcription-polymerase chain reaction (RT-PCR) assay for hantavirus using primers selected to match high homology regions of hantavirus genomes detected from the whole blood of hantavirus cardiopulmonary syndrome (HCPS) patients from Brazil, also including the N gene nucleotide sequence of Araraquara virus. Hantavirus genomes were detected in eight out of nine blood samples from the HCPS patients by RT-PCR (88.9% positivity) and in all 9 blood samples (100% positi...

  3. Recombinase Polymerase Amplification Compared to Real-Time Polymerase Chain Reaction Test for the Detection of Fasciola hepatica in Human Stool

    Science.gov (United States)

    Cabada, Miguel M.; Malaga, Jose L.; Castellanos-Gonzalez, Alejandro; Bagwell, Kelli A.; Naeger, Patrick A.; Rogers, Hayley K.; Maharsi, Safa; Mbaka, Maryann; White, A. Clinton

    2017-01-01

    Fasciola hepatica is the most widely distributed trematode infection in the world. Control efforts may be hindered by the lack of diagnostic capacity especially in remote endemic areas. Polymerase chain reaction (PCR)–based methods offer high sensitivity and specificity but require expensive technology. However, the recombinase polymerase amplification (RPA) is an efficient isothermal method that eliminates the need for a thermal cycler and has a high deployment potential to resource-limited settings. We report on the characterization of RPA and PCR tests to detect Fasciola infection in clinical stool samples with low egg burdens. The sensitivity of the RPA and PCR were 87% and 66%, respectively. Both tests were 100% specific showing no cross-reactivity with trematode, cestode, or nematode parasites. In addition, RPA and PCR were able to detect 47% and 26% of infections not detected by microscopy, respectively. The RPA adapted to a lateral flow platform was more sensitive than gel-based detection of the reaction products. In conclusion, the Fasciola RPA is a highly sensitive and specific test to diagnose chronic infection using stool samples. The Fasciola RPA lateral flow has the potential for deployment to endemic areas after further characterization. PMID:27821691

  4. [Establishment of a novel HLA genotyping method for preimplantation genetic diagnonis using multiple displacement amplification-polymerase chain reaction-sequencing based technique].

    Science.gov (United States)

    Zhang, Yinfeng; Luo, Haining; Zhang, Yunshan

    2015-12-01

    To establish a novel HLA genotyping method for preimplantation genetic diagnonis (PGD) using multiple displacement amplification-polymerase chain reaction-sequencing based technique (MDA-PCR-SBT). Peripheral blood samples and 76 1PN, 2PN, 3PN discarded embryos from 9 couples were collected. The alleles of HLA-A, B, DR loci were detected from the MDA product with the PCR-SBT method. The HLA genotypes of the parental peripheral blood samples were analyzed with the same protocol. The genotypes of specific HLA region were evaluated for distinguishing the segregation of haplotypes among the family members, and primary HLA matching was performed between the embryos. The 76 embryos were subjected to MDA and 74 (97.4%) were successfully amplified. For the 34 embryos from the single blastomere group, the amplification rate was 94.1%, and for the 40 embryos in the two blastomeres group, the rate was 100%. The dropout rates for DQ allele and DR allele were 1.3% and 0, respectively. The positive rate for MDA in the single blastomere group was 100%, with the dropout rates for DQ allele and DR allele being 1.5% and 0, respectively. The positive rate of MDA for the two blastomere group was 100%, with the dropout rates for both DQ and DR alleles being 0. The recombination rate of fetal HLA was 20.2% (30/148). Due to the improper classification and abnormal fertilized embryos, the proportion of matched embryos HLA was 20.3% (15/74),which was lower than the theoretical value of 25%. PGD with HLA matching can facilitate creation of a HLA-identical donor (saviour child) for umbilical cord blood or bone marrow stem cells for its affected sibling with a genetic disease. Therefore, preimplantation HLA matching may provide a tool for couples desiring to conceive a potential donor progeny for transplantation for its sibling with a life-threatening disorder.

  5. Miniaturized isothermal nucleic acid amplification, a review.

    Science.gov (United States)

    Asiello, Peter J; Baeumner, Antje J

    2011-04-21

    Micro-Total Analysis Systems (µTAS) for use in on-site rapid detection of DNA or RNA are increasingly being developed. Here, amplification of the target sequence is key to increasing sensitivity, enabling single-cell and few-copy nucleic acid detection. The several advantages to miniaturizing amplification reactions and coupling them with sample preparation and detection on the same chip are well known and include fewer manual steps, preventing contamination, and significantly reducing the volume of expensive reagents. To-date, the majority of miniaturized systems for nucleic acid analysis have used the polymerase chain reaction (PCR) for amplification and those systems are covered in previous reviews. This review provides a thorough overview of miniaturized analysis systems using alternatives to PCR, specifically isothermal amplification reactions. With no need for thermal cycling, isothermal microsystems can be designed to be simple and low-energy consuming and therefore may outperform PCR in portable, battery-operated detection systems in the future. The main isothermal methods as miniaturized systems reviewed here include nucleic acid sequence-based amplification (NASBA), loop-mediated isothermal amplification (LAMP), helicase-dependent amplification (HDA), rolling circle amplification (RCA), and strand displacement amplification (SDA). Also, important design criteria for the miniaturized devices are discussed. Finally, the potential of miniaturization of some new isothermal methods such as the exponential amplification reaction (EXPAR), isothermal and chimeric primer-initiated amplification of nucleic acids (ICANs), signal-mediated amplification of RNA technology (SMART) and others is presented.

  6. Development of genic SSR markers from transcriptome sequencing of pear buds.

    Science.gov (United States)

    Yue, Xiao-yan; Liu, Guo-qin; Zong, Yu; Teng, Yuan-wen; Cai, Dan-ying

    2014-04-01

    A total of 8375 genic simple sequence repeat (SSR) loci were discovered from a unigene set assembled from 116282 transcriptomic unigenes in this study. Dinucleotide repeat motifs were the most common with a frequency of 65.11%, followed by trinucleotide (32.81%). A total of 4100 primer pairs were designed from the SSR loci. Of these, 343 primer pairs (repeat length ≥15 bp) were synthesized with an M13 tail and tested for stable amplification and polymorphism in four Pyrus accessions. After the preliminary test, 104 polymorphic genic SSR markers were developed; dinucleotide and trinucleotide repeats represented 97.11% (101) of these. Twenty-eight polymorphic genic SSR markers were selected randomly to further validate genetic diversity among 28 Pyrus accessions. These markers displayed a high level of polymorphism. The number of alleles at these SSR loci ranged from 2 to 17, with a mean of 9.43 alleles per locus, and the polymorphism information content (PIC) values ranged from 0.26 to 0.91. The UPGMA (unweighted pair-group method with arithmetic average) cluster analysis grouped the 28 Pyrus accessions into two groups: Oriental pears and Occidental pears, which are congruent to the traditional taxonomy, demonstrating their effectiveness in analyzing Pyrus phylogenetic relationships, enriching rare Pyrus EST-SSR resources, and confirming the potential value of a pear transcriptome database for the development of new SSR markers.

  7. Physicochemical Properties of Avocado Pear (Persea americana ...

    African Journals Online (AJOL)

    The oil from edible avocado pear (Persea americana) was extracted using the Soxhlet extraction method and characterized for specific gravity, refractive index, free fatty acids, saponification value, iodine value, acid value and biofuel potential using standard methods. The percent oil content in the fruit pulp was determined.

  8. (Persea americana) and African pear (Dacryodes edulis)

    African Journals Online (AJOL)

    GREGO

    2007-04-02

    Apr 2, 2007 ... The percent oil content in the fruit pulp was determined. The oil content of avocado pear .... isomerisation processes which usually lead to loss of unsaturation. The low ... The role of fats and oils in cosmetics. JAOCS 49(71):.

  9. Ohmic Heating Assisted Lye Peeling of Pears.

    Science.gov (United States)

    Gupta, Sarvesh; Sastry, Sudhir K

    2018-05-01

    Currently, high concentrations (15% to 18%) of lye (sodium hydroxide) are used in peeling pears, constituting a wastewater handling and disposal problem for fruit processors. In this study, the effect of ohmic heating on lye peeling of pears was investigated. Pears were peeled using 0.5%, 1%, 2%, and 3% NaOH under different electric field strengths at two run times and their peeled yields were compared to that obtained at 2% and 18% NaOH with conventional heating. Results revealed that ohmic heating results in greater than 95% peeled yields and the best peel quality at much lower concentrations of lye (2% NaOH at 532 V/m and 3% NaOH at 426 and 479 V/m) than those obtained under conventional heating conditions. Treatment times of 30 and 60 s showed no significant differences. Within the studied range, the effects of increasing field strength yielded no significant additional benefits. These results confirm that the concentration of lye can be significantly lowered in the presence of ohmic heating to achieve high peeled yields and quality. Our work shows that lye concentrations can be greatly reduced while peeling pears, resulting in significant savings in use of caustic chemicals, reduced costs for effluent treatment and waste disposal. © 2018 Institute of Food Technologists®.

  10. Genetic variability in Brazilian populations of Biomphalaria straminea complex detected by simple sequence repeat anchored polymerase chain reaction amplification

    Directory of Open Access Journals (Sweden)

    Caldeira Roberta L

    2001-01-01

    Full Text Available Biomphalaria glabrata, B. tenagophila and B. straminea are intermediate hosts of Schistosoma mansoni, in Brazil. The latter is of epidemiological importance in the northwest of Brazil and, due to morphological similarities, has been grouped with B. intermedia and B. kuhniana in a complex named B. straminea. In the current work, we have standardized the simple sequence repeat anchored polymerase chain reaction (SSR-PCR technique, using the primers (CA8RY and K7, to study the genetic variability of these species. The similarity level was calculated using the Dice coefficient and genetic distance using the Nei and Li coefficient. The trees were obtained by the UPGMA and neighbor-joining methods. We have observed that the most related individuals belong to the same species and locality and that individuals from different localities, but of the same species, present clear heterogeneity. The trees generated using both methods showed similar topologies. The SSR-PCR technique was shown to be very efficient in intrapopulational and intraspecific studies of the B. straminea complex snails.

  11. Detection of Brazilian hantavirus by reverse transcription polymerase chain reaction amplification of N gene in patients with hantavirus cardiopulmonary syndrome

    Directory of Open Access Journals (Sweden)

    Marcos Lázaro Moreli

    2004-10-01

    Full Text Available We report a nested reverse transcription-polymerase chain reaction (RT-PCR assay for hantavirus using primers selected to match high homology regions of hantavirus genomes detected from the whole blood of hantavirus cardiopulmonary syndrome (HCPS patients from Brazil, also including the N gene nucleotide sequence of Araraquara virus. Hantavirus genomes were detected in eight out of nine blood samples from the HCPS patients by RT-PCR (88.9% positivity and in all 9 blood samples (100% positivity by nested-PCR. The eight amplicons obtained by RT-PCR (P1, P3-P9, including one obtained by nested-PCR (P-2 and not obtained by RT-PCR, were sequenced and showed high homology (94.8% to 99.1% with the N gene of Araraquara hantavirus. Although the serologic method ELISA is the most appropriate test for HCPS diagnosis, the use of nested RT-PCR for hantavirus in Brazil would contribute to the diagnosis of acute hantavirus disease detecting viral genomes in patient specimens as well as initial genomic characterization of circulating hantaviruses.

  12. Evaluation of multiplex ligation-dependent probe amplification analysis versus multiplex polymerase chain reaction assays in the detection of dystrophin gene rearrangements in an Iranian population subset

    Directory of Open Access Journals (Sweden)

    Nayereh Nouri

    2014-01-01

    Full Text Available Background: The Duchenne muscular dystrophy (DMD gene is located in the short arm of the X chromosome (Xp21. It spans 2.4 Mb of the human genomic DNA and is composed of 79 exons. Mutations in the Dystrophin gene result in DMD and Becker muscular dystrophy. In this study, the efficiency of multiplex ligation-dependent probe amplification (MLPA over multiplex polymerase chain reaction (PCR assays in an Iranian population was investigated. Materials and Methods: Multiplex PCR assays and MLPA analysis were carried out in 74 patients affected with DMD. Results: Multiplex PCR detected deletions in 51% of the patients with DMD. MLPA analysis could determine all the deletions detected by the multiplex PCR. Additionally, MLPA was able to identify one more deletion and duplication in patients without detectable mutations by multiplex PCR. Moreover, MLPA precisely determined the exact size of the deletions. Conclusion: Although MLPA analysis is more sensitive for detection of deletions and duplications in the dystrophin gene, multiplex PCR might be used for the initial analysis of the boys affected with DMD in the Iranian population as it was able to detect 95% of the rearrangements in patients with DMD.

  13. Giardia and Cryptosporidium spp. dissemination during wastewater treatment and comparative detection via immunofluorescence assay (IFA), nested polymerase chain reaction (nested PCR) and loop mediated isothermal amplification (LAMP).

    Science.gov (United States)

    Gallas-Lindemann, Carmen; Sotiriadou, Isaia; Plutzer, Judit; Noack, Michael J; Mahmoudi, Mohammad Reza; Karanis, Panagiotis

    2016-06-01

    Environmental water samples from the Lower Rhine area in Germany were investigated via immunofluorescence assays (IFAs), nested polymerase chain reaction (nested PCR) and loop-mediated isothermal amplification (LAMP) to detect the presence of Giardia spp. (n=185) and Cryptosporidium spp. (n=227). The samples were concentrated through filtration or flocculation, and oocysts were purified via centrifugation through a sucrose density gradient. For all samples, IFA was performed first, followed by DNA extraction for the nested PCR and LAMP assays. Giardia cysts were detected in 105 samples (56.8%) by IFA, 62 samples (33.5%) by nested PCR and 79 samples (42.7%) by LAMP. Cryptosporidium spp. were detected in 69 samples (30.4%) by IFA, 95 samples (41.9%) by nested PCR and 99 samples (43.6%) by LAMP. According to these results, the three detection methods are complementary for monitoring Giardia and Cryptosporidium in environmental waters. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Polymerase chain reaction amplification fails to detect aromatase cytochrome P450 transcripts in normal human endometrium or decidua.

    Science.gov (United States)

    Bulun, S E; Mahendroo, M S; Simpson, E R

    1993-06-01

    It has been proposed that the biosynthesis of estrogens by the human endometrium may be of physiological significance during the menstrual cycle. Local estrogen production was also suggested to be important in the development of endometrial cancer; however, the presence or absence of aromatase enzyme activity in normal human endometrium is controversial. To address this issue, we used a sensitive technique capable of detecting mRNA transcripts present in only very low copy number. The polymerase chain reaction linked to reverse transcription (RT-PCR) was used to evaluate the presence or absence of aromatase cytochrome P450 (P450arom) transcripts in endometrial tissues (n = 7) and endometrial stromal cells (n = 9) under various culture conditions. RNA was isolated from four proliferative and three secretory tissue samples and from cultured endometrial stromal cells isolated from seven proliferative and two secretory endometria. Five sets of cultures were treated with medroxyprogesterone acetate (MPA), estradiol (E2), and forskolin. Additionally, RNA was isolated from decidualized endometrium obtained from a patient with tubal pregnancy. A single stranded cDNA was synthesized from total RNA using Moloney murine leukemia virus reverse transcriptase and a P450arom-specific oligonucleotide. The single stranded cDNA was used as a template for PCR and was amplified for 20-35 cycles using P450arom-specific primers. RNA from adipose tissue and placenta was amplified to provide positive controls, whereas myometrial RNA was used as a negative control. In two experiments involving two endometrial tissues and three sets of cells in culture, a rat P450arom cRNA was coamplified in each sample as an internal control to demonstrate that the remote possibility of RT-PCR failures in individual test samples cannot account for our negative results. By Southern or slot blot hybridization of the amplified fragments using human and rat P450arom-specific probes, we found no evidence for

  15. Host-Guest Recognition-Assisted Electrochemical Release: Its Reusable Sensing Application Based on DNA Cross Configuration-Fueled Target Cycling and Strand Displacement Reaction Amplification.

    Science.gov (United States)

    Chang, Yuanyuan; Zhuo, Ying; Chai, Yaqin; Yuan, Ruo

    2017-08-15

    In this work, an elegantly designed host-guest recognition-assisted electrochemical release was established and applied in a reusable electrochemical biosensor for the detection of microRNA-182-5p (miRNA-182-5p), a prostate cancer biomarker in prostate cancer, based on the DNA cross configuration-fueled target cycling and strand displacement reaction (SDR) amplification. With such a design, the single target miRNA input could be converted to large numbers of single-stranded DNA (S1-Trp and S2-Trp) output, which could be trapped by cucurbit[8]uril methyl viologen (CB-8-MV 2+ ) based on the host-guest recognition, significantly enhancing the sensitivity for miRNA detection. Moreover, the nucleic acids products obtained from the process of cycling amplification could be utilized sufficiently, avoiding the waste and saving the experiment cost. Impressively, by resetting a settled voltage, the proposed biosensor could release S1-Trp and S2-Trp from the electrode surface, attributing that the guest ion methyl viologen (MV 2+ ) was reduced to MV +· under this settled voltage and formed a more-stable CB-8-MV +· -MV +· complex. Once O 2 was introduced in this system, MV +· could be oxidized to MV 2+ , generating the complex of CB-8-MV 2+ for capturing S1-Trp and S2-Trp again in only 5 min. As a result, the simple and fast regeneration of biosensor for target detection was realized on the base of electrochemical redox-driven assembly and release, overcoming the challenges of time-consuming, burdensome operations and expensive experimental cost in traditional reusable biosensors and updating the construction method for a reusable bisensor. Furthermore, the biosensor could be reused for more than 10 times with a regeneration rate of 93.20%-102.24%. After all, the conception of this work provides a novel thought for the construction of effective reusable biosensor to detect miRNA and other biomarkers and has great potential application in the area requiring the release of

  16. Detection of influenza viruses by coupling multiplex reverse-transcription loop-mediated isothermal amplification with cascade invasive reaction using nanoparticles as a sensor

    Directory of Open Access Journals (Sweden)

    Ge Y

    2017-04-01

    Full Text Available Yiyue Ge,1 Qiang Zhou,2 Kangchen Zhao,1 Ying Chi,1 Bin Liu,3 Xiaoyan Min,4 Zhiyang Shi,1 Bingjie Zou,2 Lunbiao Cui1 1Institute of Pathogenic Microbiology, Key Laboratories of Enteric Pathogenic Microbiology (Ministry of Health, Jiangsu Provincial Center for Disease Control and Prevention, 2Department of Pharmacology, Jinling Hospital, Medical School of Nanjing University, 3Department of Biomedical Engineering, Nanjing Medical University, 4Department of Geriatrics, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, People’s Republic of China Abstract: Influenza virus infections represent a worldwide public health and economic problem due to the significant morbidity and mortality caused by seasonal epidemics and pandemics. Sensitive and convenient methodologies for detection of influenza viruses are essential for further disease control. Loop-mediated isothermal amplification (LAMP is the most commonly used method of nucleic acid isothermal amplification. However, with regard to multiplex LAMP, differentiating the ladder-like LAMP products derived from multiple targets is still challenging today. The requirement of specialized instruments has further hindered the on-site application of multiplex LAMP. We have developed an integrated assay coupling multiplex reverse transcription LAMP with cascade invasive reaction using nanoparticles (mRT-LAMP-CIRN as a sensor for the detection of three subtypes of influenza viruses: A/H1N1pdm09, A/H3 and influenza B. The analytic sensitivities of the mRT-LAMP-CIRN assay were 101 copies of RNA for both A/H1N1pdm09 and A/H3, and 102 copies of RNA for influenza B. This assay demonstrated highly specific detection of target viruses and could differentiate them from other genetically or clinically related viruses. Clinical specimen analysis showed the mRT-LAMP-CIRN assay had an overall sensitivity and specificity of 98.3% and 100%, respectively. In summary, the mRT-LAMP-CIRN assay is

  17. Development of a multiplex amplification refractory mutation system reverse transcription polymerase chain reaction assay for the differential diagnosis of Feline leukemia virus vaccine and wild strains.

    Science.gov (United States)

    Ho, Chia-Fang; Chan, Kun-Wei; Yang, Wei-Cheng; Chiang, Yu-Chung; Chung, Yang-Tsung; Kuo, James; Wang, Chi-Young

    2014-07-01

    A multiplex amplification refractory mutation system reverse transcription polymerase chain reaction (ARMS RT-PCR) was developed for the differential diagnosis of Feline leukemia virus (FeLV) vaccine and wild-type strains based on a point mutation between the vaccine strain (S) and the wild-type strain (T) located in the p27 gene. This system was further upgraded to obtain a real-time ARMS RT-PCR (ARMS qRT-PCR) with a high-resolution melt analysis (HRMA) platform. The genotyping of various strains of FeLV was determined by comparing the HRMA curves with the defined wild-type FeLV (strain TW1), and the results were expressed as a percentage confidence. The detection limits of ARMS RT-PCR and ARMS qRT-PCR combined with HRMA were 100 and 1 copies of transcribed FeLV RNA per 0.5 ml of sample, respectively. No false-positive results were obtained with 6 unrelated pathogens and 1 feline cell line. Twelve FeLV Taiwan strains were correctly identified using ARMS qRT-PCR combined with HRMA. The genotypes of the strains matched the defined FeLV wild-type strain genotype with at least 91.17% confidence. A higher degree of sequence polymorphism was found throughout the p27 gene compared with the long terminal repeat region. In conclusion, the current study describes the phylogenetic relationship of the FeLV Taiwan strains and demonstrates that the developed ARMS RT-PCR assay is able to be used to detect the replication of a vaccine strain that has not been properly inactivated, thus acting as a safety check for the quality of FeLV vaccines.

  18. Development of a multiplex polymerase chain reaction-sequence-specific primer method for NKG2D and NKG2F single-nucleotide polymorphism typing using isothermal multiple displacement amplification products.

    Science.gov (United States)

    Kaewmanee, M; Phoksawat, W; Romphruk, A; Romphruk, A V; Jumnainsong, A; Leelayuwat, C

    2013-06-01

    Natural killer group 2 member D (NKG2D) on immune effector cells recognizes multiple stress-inducible ligands. NKG2D single-nucleotide polymorphism (SNP) haplotypes were related to the levels of cytotoxic activity of peripheral blood mononuclear cells. Indeed, these polymorphisms were also located in NKG2F. Isothermal multiple displacement amplification (IMDA) is used for whole genome amplification (WGA) that can amplify very small genomic DNA templates into microgram with whole genome coverage. This is particularly useful in the cases of limited amount of valuable DNA samples requiring multi-locus genotyping. In this study, we evaluated the quality and applicability of IMDA to genetic studies in terms of sensitivity, efficiency of IMDA re-amplification and stability of IMDA products. The smallest amount of DNA to be effectively amplified by IMDA was 200 pg yielding final DNA of approximately 16 µg within 1.5 h. IMDA could be re-amplified only once (second round of amplification), and could be kept for 5 months at 4°C and more than a year at -20°C without loosing genome coverage. The amplified products were used successfully to setup a multiplex polymerase chain reaction-sequence-specific primer for SNP typing of the NKG2D/F genes. The NKG2D/F multiplex polymerase chain reaction (PCR) contained six PCR mixtures for detecting 10 selected SNPs, including 8 NKG2D/F SNP haplotypes and 2 additional NKG2D coding SNPs. This typing procedure will be applicable in both clinical and research laboratories. Thus, our data provide useful information and limitations for utilization of genome-wide amplification using IMDA and its application for multiplex NKG2D/F typing. © 2013 John Wiley & Sons Ltd.

  19. Characterization and analysis of CCR and CAD gene families at the whole-genome level for lignin synthesis of stone cells in pear (Pyrus bretschneideri) fruit.

    Science.gov (United States)

    Cheng, Xi; Li, Manli; Li, Dahui; Zhang, Jinyun; Jin, Qing; Sheng, Lingling; Cai, Yongping; Lin, Yi

    2017-11-15

    The content of stone cells has significant effects on the flavour and quality of pear fruit. Previous research suggested that lignin deposition is closely related to stone cell formation. In the lignin biosynthetic pathway, cinnamoyl-CoA reductase (CCR) and cinnamyl alcohol dehydrogenase (CAD), dehydrogenase/reductase family members, catalyse the last two steps in monolignol synthesis. However, there is little knowledge of the characteristics of the CCR and CAD families in pear and their involvement in lignin synthesis of stone cells. In this study, 31 CCR s and 26 CAD s were identified in the pear genome. Phylogenetic trees for CCR s and CAD s were constructed; key amino acid residues were analysed, and three-dimensional structures were predicted. Using quantitative real-time polymerase chain reaction (qRT-PCR), PbCAD2 , PbCCR1 , -2 and - 3 were identified as participating in lignin synthesis of stone cells in pear fruit. Subcellular localization analysis showed that the expressed proteins (PbCAD2, PbCCR1, -2 and -3) are found in the cytoplasm or at the cell membrane. These results reveal the evolutionary features of the CCR and CAD families in pear as well as the genes responsible for regulation of lignin synthesis and stone cell development in pear fruit. © 2017. Published by The Company of Biologists Ltd.

  20. Characterization and analysis of CCR and CAD gene families at the whole-genome level for lignin synthesis of stone cells in pear (Pyrus bretschneideri fruit

    Directory of Open Access Journals (Sweden)

    Xi Cheng

    2017-11-01

    Full Text Available The content of stone cells has significant effects on the flavour and quality of pear fruit. Previous research suggested that lignin deposition is closely related to stone cell formation. In the lignin biosynthetic pathway, cinnamoyl-CoA reductase (CCR and cinnamyl alcohol dehydrogenase (CAD, dehydrogenase/reductase family members, catalyse the last two steps in monolignol synthesis. However, there is little knowledge of the characteristics of the CCR and CAD families in pear and their involvement in lignin synthesis of stone cells. In this study, 31 CCRs and 26 CADs were identified in the pear genome. Phylogenetic trees for CCRs and CADs were constructed; key amino acid residues were analysed, and three-dimensional structures were predicted. Using quantitative real-time polymerase chain reaction (qRT-PCR, PbCAD2, PbCCR1, -2 and -3 were identified as participating in lignin synthesis of stone cells in pear fruit. Subcellular localization analysis showed that the expressed proteins (PbCAD2, PbCCR1, -2 and -3 are found in the cytoplasm or at the cell membrane. These results reveal the evolutionary features of the CCR and CAD families in pear as well as the genes responsible for regulation of lignin synthesis and stone cell development in pear fruit.

  1. Effect of hormone treatments on deformed fruit development in pear

    African Journals Online (AJOL)

    erica

    2012-05-31

    May 31, 2012 ... of plant growth regulators applications and endogenous hormones on deformed pear fruit to clarify the relation- ship between shape and hormones in the fruit. MATERIALS AND METHODS. The experiments were conducted in 2008 using pear trees growing fruit demonstration farm in Jianning County.

  2. 76 FR 78168 - Importation of Chinese Sand Pears From China

    Science.gov (United States)

    2011-12-16

    ... require the sand pears to be packed in cartons that are labeled with the identity of the place of... of China. (5) The pears must be packed in cartons that are labeled with the identity of the place of... registration of places of production and packinghouses, sourcing of pest-free propagative material, inspection...

  3. Changes in endogenous hormone contents of pear stock ( Pyrus ...

    African Journals Online (AJOL)

    In this study, changes in endogenous hormone contents of pear stock seeds during cold stratification were investigated. Abscisic acid (ABA) content decreased with increase in the periods of stratification of pear stock seeds. However, gibberellic acid (GA) and indole-3-acetic acid (IAA) contents of Pyrus betulaefolia and ...

  4. Detection of pear thrips damage using satellite imagery data

    Science.gov (United States)

    James E. Vogelmann; Barrett N. Rock

    1991-01-01

    This study evaluates the potential of measuring, mapping and monitoring sugar maple damage caused by pear thrips in southern Vermont and northwestern Massachusetts using satellite imagery data. Landsat Thematic Mapper (TM) data were obtained during a major thrips infestation in June 1988, and were compared with satellite data acquired during June 1984 (before pear...

  5. 7 CFR 457.111 - Pear crop insurance provisions.

    Science.gov (United States)

    2010-01-01

    ... alternating or mixed pattern. Marketable. Pear production acceptable for processing or other human consumption... irrigation water supply, if caused by an insured peril that occurs during the insurance period. (b) In... provisions of the Basic Provisions are not applicable. 13. Pear Quality Adjustment Endorsement (a) This...

  6. Wet faeces produced by sheep fed dried spineless cactus pear ...

    African Journals Online (AJOL)

    Cactus pear cladodes in ruminant diets are characterized by the production of wet faeces and assumed to be diarrhoea. Incremental levels of sun-dried and coarsely ground spineless cactus pear (Opuntia ficus-indica var. Algerian) cladodes were used to substitute part of the lucerne hay in balanced sheep diets. Feed and ...

  7. 7 CFR 917.461 - Pear Regulation 12.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false Pear Regulation 12. 917.461 Section 917.461 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing... grower to the field or area in which the pears are grown for 12 months prior to the appearance of flower...

  8. Quality characteristics and phenolic compounds of European pear ...

    African Journals Online (AJOL)

    Background: Pear fruits are an important source of plant secondary metabolites and one of the major sources of dietary phenolic compounds. Materials and Methods: The aim of this study was to determine the individual phenolic compounds and some quality characteristics of the flesh and peel of the fruit in four pear ...

  9. DNAzyme Feedback Amplification: Relaying Molecular Recognition to Exponential DNA Amplification.

    Science.gov (United States)

    Liu, Meng; Yin, Qingxin; McConnell, Erin M; Chang, Yangyang; Brennan, John D; Li, Yingfu

    2018-03-26

    Technologies capable of linking DNA amplification to molecular recognition are very desirable for ultrasensitive biosensing applications. We have developed a simple but powerful isothermal DNA amplification method, termed DNAzyme feedback amplification (DFA), that is capable of relaying molecular recognition to exponential DNA amplification. The method incorporates both an RNA-cleaving DNAzyme (RCD) and rolling circle amplification (RCA) carried out by a special DNA polymerase using a circular DNA template. DFA begins with a stimulus-dependent RCA reaction, producing tandemly linked RCDs in long-chain DNA products. These RCDs cleave an RNA-containing DNA sequence to form additional primers that hybridize to the circular DNA molecule, giving rise to DNA assemblies that act as the new inputs for RCA. The RCA reaction and the cleavage event keep on feeding each other autonomously, resulting in exponential growth of repetitive DNA sequences that can be easily detected. This method can be used for the detection of both nucleic acid based targets and non-nucleic acid analytes. In this article, we discuss the conceptual framework of the feedback amplification approach, the essential features of this method as well as remaining challenges and possible solutions. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Pear distillates from pear juice concentrate: effect of lees in the aromatic composition.

    Science.gov (United States)

    García-Llobodanin, L; Achaerandio, I; Ferrando, M; Güell, C; López, F

    2007-05-02

    Pear juice obtained from pear concentrate was fermented at room temperature using Saccharomyces cerevisiae (BDX, ENOFERM, France) as the fermentation microorganism. During the fermentation process, total sugars were measured. High performance liquid chromatography analyses were used to monitor the fermentation process and to characterize the pear wine. The pear wine obtained was distilled with its lees using three different equipments: a glass alembic (a glass pot still coupled to a glass column), a copper alembic, and a glass alembic with the addition of 5 g/L of copper shavings to the pot still. The same distillations were repeated with the wine without its lees (separated by decanting). Several distillation fractions were collected, up to a total of 500 mL of distillate. Gas chromatography was used to identify and quantify the volatile compounds in each fraction, and the methanol and ethanol contents. Based on these results, the heart fraction was defined. ANOVA tests were performed on the heart fractions to determine quantitative differences between some volatile compounds depending on the equipment used and the presence or absence of the wine lees. From this series of ANOVA tests, it can be concluded that the concentrations of the compounds that are considered to have a negative effect on the quality of the distillates (methanol, ethyl acetate, furfural) decrease or do not change when they are distilled in the presence of lees and in the copper alembic. In addition, the concentrations of the positive compounds (ethyl decanoate and ethyl-2-trans-4-cis-decadienoate) increase in the presence of lees for all of the equipment tested. So, it can be assumed that the distillation of pear wine with its lees in copper alembic leads to a better quality product.

  11. Adsorption isotherms of pear at several temperatures

    OpenAIRE

    Mitrevski Vangelče; Lutovska Monika; Mijakovski Vladimir; Pavkov Ivan S.; Babić Mirko M.; Radojčin Milivoje T.

    2015-01-01

    The moisture adsorption isotherms of pear were determined at 15ºC, 30ºC and 45ºC using the standard static gravimetric method over a range of water activity from 0.112 to 0.920. The experimental data were fitted with isotherm equations recommended in ASAE Standard D245.5. In order to find which equation gives the best results, large number of numerical experiments were performed. After that, several statistical criteria proposed in scientific literature for...

  12. Pear transformed with a lytic peptide gene for disease control affects nontarget organism, pear psylla (Homoptera: Psyllidae).

    Science.gov (United States)

    Puterka, Gary J; Bocchetti, Chris; Dang, Phat; Bell, R L; Scorza, Ralph

    2002-08-01

    The biology and behavior of pear psylla, Cacopsylla pyricola Foerster, on a transgenic clone of 'Bartlett' pear, Pyrus communis L., containing a synthetic antimicrobial gene, D5C1, was compared with that of a nontransgenic parental clone to determine whether there were any nontarget effects. The gene construct also contained the marker gene nptII (aminoglycoside 3'-phosphotransferase II) that encodes for antibiotic resistance to identify transformed plants. The purpose of the original transformation was to enhance pear resistance to the bacterial disease fireblight caused by Erwinia amylovora (Burr.) Winslow et al. The biology and behavior of pear psylla on a transgenic clone were compared with a nontransgenic parental pear clone in short- (crops.

  13. Use of FTA elute card impregnated with cervicovaginal sample directly into the amplification reaction increases the detection of human papillomavirus DNA.

    Science.gov (United States)

    Santos, Carla R; Franciscatto, Laura G; Barcellos, Regina B; Almeida, Sabrina E M; Rossetti, Maria Lucia R

    2012-01-01

    This study aimed to evaluate the use of the FTA elute card(TM) impregnated with cervicovaginal sample directly in the PCR amplification for detection of HPV-DNA. The results were compared to a reference technique. This method was more efficient than the protocol indicated by the manufacturer, identifying 91.7% against 54.2% of the positive samples.

  14. Use of FTA elute card impregnated with cervicovaginal sample directly into the amplification reaction increases the detection of human papillomavirus DNA

    OpenAIRE

    Santos, Carla R.; Franciscatto, Laura G.; Barcellos, Regina B.; Almeida, Sabrina E. M; Rossetti, Maria Lucia R.

    2012-01-01

    This study aimed to evaluate the use of the FTA elute cardTM impregnated with cervicovaginal sample directly in the PCR amplification for detection of HPV-DNA. The results were compared to a reference technique. This method was more efficient than the protocol indicated by the manufacturer, identifying 91.7% against 54.2% of the positive samples.

  15. Use of FTA elute card impregnated with cervicovaginal sample directly into the amplification reaction increases the detection of human papillomavirus DNA

    Directory of Open Access Journals (Sweden)

    Carla R. Santos

    2012-03-01

    Full Text Available This study aimed to evaluate the use of the FTA elute cardTM impregnated with cervicovaginal sample directly in the PCR amplification for detection of HPV-DNA. The results were compared to a reference technique. This method was more efficient than the protocol indicated by the manufacturer, identifying 91.7% against 54.2% of the positive samples.

  16. Mutant of Japanese pear resistant to Black Spot Disease

    International Nuclear Information System (INIS)

    Sanada, T.; Nishida, T.; Ikeda, F.

    1987-01-01

    Full text: Nijisseike is one of the leading cultivars of Japanese pear (Pyrus serotinea Rehd.), but susceptible to black spot disease. Farmers try to prevent this disease by wrapping the fruit with a paper bag and by repeated spraying of fungicides. The disease is caused by a Japanese pear pathotype of Alternaria alternata (Fr.) Keissler. Susceptibility is controlled by a single dominant gene. In 1962, grafted trees of this cultivar were planted at a distance between 53 and 93 m from the 60 Co source in the gamma-field (daily dose 15-4 rad). One branch on a tree planted at 53 m was detected as resistant in 1981. Under field conditions, black spots were observed on many fruits and leaves of the original trees by natural infection in early July, however, they were not observed on the mutant. To examine the resistance of the mutant, artificial inoculations were made using spores of the pathogen and the host specific toxin produced by germinating spores. When some drops of the spore suspension are placed on leaves, the formation of black spots depends upon the leaf age. In a resistant cv. as Chojuro, black spot symptoms are formed only when inoculated on young leaves. An intermediate reaction was observed in the mutant, whereas the original Nijisseiki showed severe symptoms. When inoculation was made on matured fruit skins, no black spot was formed on the mutant just like on the resistant cv. Chojuro, while many small black spots were formed and grew into large spots overlapping each other on the susceptible cv. Nijisseiki. In case of the crude toxin inoculation (4-0.04 ppm) of cv. Nijisseiki black spots were formed on the surface of the susceptible fruit skin, and necrotic lesions at the cut end of detached small pieces of leaves, although reaction on fruit skins was weaker compared with inoculation by spores. However, no symptoms were observed from the toxin application on the mutant and the resistant cv. Chojuro. That the resistance of the mutant is classified as

  17. Subject in Tractatus according to David Pears

    Directory of Open Access Journals (Sweden)

    M Hoseinzadeh Yazdi

    2013-02-01

    Full Text Available Subjectivism is viewed as one of the most fundamental underpinnings of modern philosophy. In modern philosophy, subject takes up a new position in human knowledge. The formation of the concept of subject is a decisive turn with which the modern philosophy starts. Considering the centrality of subjectivism in modern philosophy, this article attempts to explain subject in Tractatus according to David Pears. A review of Wittgenstein’s earlier teachings reveals that he considers a fundamental limitation for language. The subject serves as a point of view from which the language can be understood. The subject is the presupposition of understanding. Another way of putting this would be to say that any experience is understood from a point of view which is not represented in that experience. Regarding this, it seems that earlier Wittgenstein is somehow subjectivist. This specific form of subjectivism is different from Kantian subjectivism.

  18. Genomics of pear and other Rosaceae fruit trees.

    Science.gov (United States)

    Yamamoto, Toshiya; Terakami, Shingo

    2016-01-01

    The family Rosaceae includes many economically important fruit trees, such as pear, apple, peach, cherry, quince, apricot, plum, raspberry, and loquat. Over the past few years, whole-genome sequences have been released for Chinese pear, European pear, apple, peach, Japanese apricot, and strawberry. These sequences help us to conduct functional and comparative genomics studies and to develop new cultivars with desirable traits by marker-assisted selection in breeding programs. These genomics resources also allow identification of evolutionary relationships in Rosaceae, development of genome-wide SNP and SSR markers, and construction of reference genetic linkage maps, which are available through the Genome Database for the Rosaceae website. Here, we review the recent advances in genomics studies and their practical applications for Rosaceae fruit trees, particularly pear, apple, peach, and cherry.

  19. Aerial spray trials for pear thrips management fall 1988

    Science.gov (United States)

    H. Brenton Teillon; Bruce L. Parker

    1991-01-01

    The defoliation from pear thrips, Taeniothrips inconsequens (Uzel), in 1988 caused a great deal of public concern throughout the entire State of Vermont and the New England region (Parker et al. 1988).

  20. Changing perspectives on pearly mussels, North America's most imperiled animals

    Science.gov (United States)

    Strayer, David L.; Downing, John A.; Haag, Wendell R.; King, Timothy L.; Layzer, James B.; Newton, Teresa J.; Nichols, S. Jerrine

    2004-01-01

    Pearly mussels (Unionacea) are widespread, abundant, and important in freshwater ecosystems around the world. Catastrophic declines in pearly mussel populations in North America and other parts of the world have led to a flurry of research on mussel biology, ecology, and conservation. Recent research on mussel feeding, life history, spatial patterning, and declines has augmented, modified, or overturned long-held ideas about the ecology of these animals. Pearly mussel research has begun to benefit from and contribute to current ideas about suspension feeding, life-history theory, metapopulations, flow refuges, spatial patterning and its effects, and management of endangered species. At the same time, significant gaps in understanding and apparent paradoxes in pearly mussel ecology have been exposed. To conserve remaining mussel populations, scientists and managers must simultaneously and aggressively pursue both rigorous research and conservation actions.

  1. The genome of the pear (Pyrus bretschneideri Rehd.).

    Science.gov (United States)

    Wu, Jun; Wang, Zhiwen; Shi, Zebin; Zhang, Shu; Ming, Ray; Zhu, Shilin; Khan, M Awais; Tao, Shutian; Korban, Schuyler S; Wang, Hao; Chen, Nancy J; Nishio, Takeshi; Xu, Xun; Cong, Lin; Qi, Kaijie; Huang, Xiaosan; Wang, Yingtao; Zhao, Xiang; Wu, Juyou; Deng, Cao; Gou, Caiyun; Zhou, Weili; Yin, Hao; Qin, Gaihua; Sha, Yuhui; Tao, Ye; Chen, Hui; Yang, Yanan; Song, Yue; Zhan, Dongliang; Wang, Juan; Li, Leiting; Dai, Meisong; Gu, Chao; Wang, Yuezhi; Shi, Daihu; Wang, Xiaowei; Zhang, Huping; Zeng, Liang; Zheng, Danman; Wang, Chunlei; Chen, Maoshan; Wang, Guangbiao; Xie, Lin; Sovero, Valpuri; Sha, Shoufeng; Huang, Wenjiang; Zhang, Shujun; Zhang, Mingyue; Sun, Jiangmei; Xu, Linlin; Li, Yuan; Liu, Xing; Li, Qingsong; Shen, Jiahui; Wang, Junyi; Paull, Robert E; Bennetzen, Jeffrey L; Wang, Jun; Zhang, Shaoling

    2013-02-01

    The draft genome of the pear (Pyrus bretschneideri) using a combination of BAC-by-BAC and next-generation sequencing is reported. A 512.0-Mb sequence corresponding to 97.1% of the estimated genome size of this highly heterozygous species is assembled with 194× coverage. High-density genetic maps comprising 2005 SNP markers anchored 75.5% of the sequence to all 17 chromosomes. The pear genome encodes 42,812 protein-coding genes, and of these, ~28.5% encode multiple isoforms. Repetitive sequences of 271.9 Mb in length, accounting for 53.1% of the pear genome, are identified. Simulation of eudicots to the ancestor of Rosaceae has reconstructed nine ancestral chromosomes. Pear and apple diverged from each other ~5.4-21.5 million years ago, and a recent whole-genome duplication (WGD) event must have occurred 30-45 MYA prior to their divergence, but following divergence from strawberry. When compared with the apple genome sequence, size differences between the apple and pear genomes are confirmed mainly due to the presence of repetitive sequences predominantly contributed by transposable elements (TEs), while genic regions are similar in both species. Genes critical for self-incompatibility, lignified stone cells (a unique feature of pear fruit), sorbitol metabolism, and volatile compounds of fruit have also been identified. Multiple candidate SFB genes appear as tandem repeats in the S-locus region of pear; while lignin synthesis-related gene family expansion and highly expressed gene families of HCT, C3'H, and CCOMT contribute to high accumulation of both G-lignin and S-lignin. Moreover, alpha-linolenic acid metabolism is a key pathway for aroma in pear fruit.

  2. Carbon footprint of apple and pear : orchards, storage and distribution

    OpenAIRE

    Figueiredo, F.; Castanheira, E.G.; Feliciano, M.; Rodrigues, M.A.; Peres, A.; Maia, F.; Ramos, A.; Carneiro, J.P.; Coroama, V.C.; Freire, F.

    2013-01-01

    Apple and pear represent 51% of fresh fruit orchards in Portugal. This paper presents a life-cycle (LC) greenhouse gas (GHG) assessment (so-called carbon footprint) of 3 apple and 1 pear Portuguese production systems. An LC model and inventory were implemented, encompassing the farm stage (cultivation of fruit trees in orchards), storage and distribution (transport to retail). The functional unit considered in this study was 1 kg of distributed fruit (at retail). Four different LC inventories...

  3. Amplification factor variable amplifier

    NARCIS (Netherlands)

    Akitsugu, Oshita; Nauta, Bram

    2007-01-01

    PROBLEM TO BE SOLVED: To provide an amplification factor variable amplifier capable of achieving temperature compensation of an amplification factor over a wide variable amplification factor range. ; SOLUTION: A Gilbert type amplification factor variable amplifier 11 amplifies an input signal and

  4. Amplification factor variable amplifier

    NARCIS (Netherlands)

    Akitsugu, Oshita; Nauta, Bram

    2010-01-01

    PROBLEM TO BE SOLVED: To provide an amplification factor variable amplifier capable of achieving temperature compensation of an amplification factor over a wide variable amplification factor range. ;SOLUTION: A Gilbert type amplification factor variable amplifier 11 amplifies an input signal and can

  5. Diagnostic Devices for Isothermal Nucleic Acid Amplification

    Directory of Open Access Journals (Sweden)

    Chia-Chen Chang

    2012-06-01

    Full Text Available Since the development of the polymerase chain reaction (PCR technique, genomic information has been retrievable from lesser amounts of DNA than previously possible. PCR-based amplifications require high-precision instruments to perform temperature cycling reactions; further, they are cumbersome for routine clinical use. However, the use of isothermal approaches can eliminate many complications associated with thermocycling. The application of diagnostic devices for isothermal DNA amplification has recently been studied extensively. In this paper, we describe the basic concepts of several isothermal amplification approaches and review recent progress in diagnostic device development.

  6. Diagnostic devices for isothermal nucleic acid amplification.

    Science.gov (United States)

    Chang, Chia-Chen; Chen, Chien-Cheng; Wei, Shih-Chung; Lu, Hui-Hsin; Liang, Yang-Hung; Lin, Chii-Wann

    2012-01-01

    Since the development of the polymerase chain reaction (PCR) technique, genomic information has been retrievable from lesser amounts of DNA than previously possible. PCR-based amplifications require high-precision instruments to perform temperature cycling reactions; further, they are cumbersome for routine clinical use. However, the use of isothermal approaches can eliminate many complications associated with thermocycling. The application of diagnostic devices for isothermal DNA amplification has recently been studied extensively. In this paper, we describe the basic concepts of several isothermal amplification approaches and review recent progress in diagnostic device development.

  7. 不同品种梨果石细胞含量差异研究%Study on the Difference of Stone Cells Content in Different Pear Varieties

    Institute of Scientific and Technical Information of China (English)

    黄凯; 王娟娟; 刘汉云

    2017-01-01

    为了探索不同梨品种果实的石细胞差异,以硕丰梨、玉露香梨、巴梨为试材,测定了3个品种梨果实石细胞含量、石细胞团的分布、石细胞团的大小与密度等指标.结果表明,石细胞含量由高到低依次为硕丰梨>玉露香梨>巴梨,果实不同部位石细胞含量由高到低依次为近果心>近果皮>果肉中部;同一品种的梨果,果实较小的梨果石细胞含量更高.间苯三酚-盐酸染色结果显示,石细胞团主要分布在果心处,且呈现出辐射状由果心向果肉中延伸,分布特点是近果心>近果皮>果肉中部.近果心处石细胞分布密度由高到低依次为硕丰梨>玉露香梨>巴梨.研究结果可为改善梨果实品质奠定基础.%To explore the difference of stone cell,‘Yuluxiang'pear,Bartlett pear and ‘Shuofeng'pear were chose as the research object,the paper mainly measured the indexes of the stone cell content,the distribution of stone cell and the density and size of stone cell.The results showed that the order (from higher to lower) of the stone cell content in different pear varieties was ‘Shuofeng'pear >‘Yuluxiang'pear > Bartlett pear.The order (from higher to lower) of the stone cell content in different parts was core region > peel region > middle part.The smaller pear had a higher content of stone cell in one variety.The result of Wiesner reaction showed that the group of stone cells mainly distributed in the core,and showed a radiation shape from the core to the pulp extends.The distribution characteristics was the stone ceils in the core > the stone cells near the pear peel > the stone cells in the middle flesh.The density distribution of the stone cell in the core paris was ‘Shuofeng'pear > ‘Yuluxiang'pear > Bartlett pear.This research laid the foundation for improving the quality of pear fruit.

  8. Adsorption isotherms of pear at several temperatures

    Directory of Open Access Journals (Sweden)

    Mitrevski Vangelče

    2015-01-01

    Full Text Available The moisture adsorption isotherms of pear were determined at 15ºC, 30ºC and 45ºC using the standard static gravimetric method over a range of water activity from 0.112 to 0.920. The experimental data were fitted with isotherm equations recommended in ASAE Standard D245.5. In order to find which equation gives the best results, large number of numerical experiments were performed. After that, several statistical criteria proposed in scientific literature for estimation and selection of the best sorption isotherm equations were used. For each equation and experimental data set, the average performance index was calculated and models were ranked afterwards. After that, some statistical rejection criteria were checked (D’Agostino-Pearson test of normality, single-sample run test and significance and precision of the model parameters. The performed statistical analysis shows that the Guggenheim-Anderson-de Boer (GAB equation has the highest value of average performance index, but higher correlation between pair of parameters leads to lower precision of estimated parameters.[Projekat Ministarstva nauke Republike Srbije, br. TR 31058

  9. Genetic linkage maps of Japanese and European pears aligned to the apple consensus map

    NARCIS (Netherlands)

    Yamamoto, T.; Kimura, T.; Saito, T.; Kotobuki, K.; Matsuta, N.; Liebhard, R.; Gessler, C.; Weg, van de W.E.; Hayashi, T.

    2004-01-01

    Genetic linkage maps of the Japanese pear (Pyrus pyrifolia Nakai) cultivar `Housui¿ and the European pear (Pyrus communis L.) cultivar `Bartlett¿ were constructed based on Amplified Fragment Length Polymorphism markers (AFLPs), Simple Sequence Repeat markers (SSRs) (from pear, apple and Prunus),

  10. 7 CFR 319.56-20 - Apples and pears from Australia (including Tasmania) and New Zealand.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 5 2010-01-01 2010-01-01 false Apples and pears from Australia (including Tasmania... Fruits and Vegetables § 319.56-20 Apples and pears from Australia (including Tasmania) and New Zealand. Apples and pears from Australia (including Tasmania) and New Zealand may be imported only in accordance...

  11. Origin, Domestication, and Dispersing of Pear (Pyrus spp.

    Directory of Open Access Journals (Sweden)

    G. J. Silva

    2014-01-01

    Full Text Available The pear (Pyrus communis L. is a typical fruit of temperate regions, having its origin and domestication at two different points, China and Asia Minor until the Middle East. It is the fifth most widely produced fruit in the world, being produced mainly in China, Europe, and the United States. Pear belongs to rosaceous family, being a close “cousin” of the apple, but with some particularities that make this fruit special with a delicate flavor. Thus, it deserves a special attention and a meticulous review of all the history involved, and the recent research devoted to it, because of the economic and cultural importance of this fruit in a range of countries and cultures. Therefore, the purpose of this literature review is to approach the history of the origin, domestication, and dispersal of pears, as well as reporting their botany, their current scenario in the world, and their breeding and conservation.

  12. Nutrient removal by apple, pear and cherry nursery trees

    OpenAIRE

    Giovambattista Sorrenti; Maurizio Quartieri; Silvia Salvi; Moreno Toselli

    2017-01-01

    Given that nursery is a peculiar environment, the amount of nutrients removed by nursery trees represents a fundamental acquisition to optimise fertilisation strategies, with economic and environmental implications. In this context, we determined nutrient removal by apple, pear and cherry nursery trees at the end of the nursery growing cycle. We randomly removed 5 leafless apple (Golden Delicious/EMLA M9; density of 30,000 trees ha–1), pear (Santa Maria/Adams; density of 30,000 trees ha–1) an...

  13. Utility of loop-mediated isothermal amplification assay, polymerase chain reaction, and elisa for diagnosis of leptospirosis in South Indian patients

    Directory of Open Access Journals (Sweden)

    Mallika Sengupta

    2017-01-01

    Full Text Available Background: Leptospirosis is a zoonotic disease which requires laboratory diagnosis for confirmation. Materials and Methods: In this study serum samples from adults with acute undifferentiated fever (duration ≤15 days were tested for IgM antibodies to Leptospira by ELISA, PCR for rrs gene and loop-mediated isothermal amplification (LAMP assay for LipL32 and LipL41. Results: Among the 150 sera tested, three were positive by PCR, LAMP and IgM ELISA/modified Faines' criteria, two by only PCR; seven only by LAMP assay and forty fulfilled modified Faine's criteria (illness clinically compatible and IgM ELISA positive for leptospirosis. Clinical correlation revealed renal compromise, low platelet count and severe jaundice were significantly related to leptospirosis (P < 0.05. Conclusion: This study suggests that LAMP assay could be useful for diagnosis of leptospirosis during the 1st week of illness whereas IgM ELISA forms the mainstay of diagnosis from the 2nd week onward. Further studies especially community based, comparing ELISA, PCR, LAMP, culture and microscopic agglutination test are required to evaluate the veracity of these findings.

  14. Helicase-dependent amplification of nucleic acids.

    Science.gov (United States)

    Cao, Yun; Kim, Hyun-Jin; Li, Ying; Kong, Huimin; Lemieux, Bertrand

    2013-10-11

    Helicase-dependent amplification (HDA) is a novel method for the isothermal in vitro amplification of nucleic acids. The HDA reaction selectively amplifies a target sequence by extension of two oligonucleotide primers. Unlike the polymerase chain reaction (PCR), HDA uses a helicase enzyme to separate the deoxyribonucleic acid (DNA) strands, rather than heat denaturation. This allows DNA amplification without the need for thermal cycling. The helicase used in HDA is a helicase super family II protein obtained from a thermophilic organism, Thermoanaerobacter tengcongensis (TteUvrD). This thermostable helicase is capable of unwinding blunt-end nucleic acid substrates at elevated temperatures (60° to 65°C). The HDA reaction can also be coupled with reverse transcription for ribonucleic acid (RNA) amplification. The products of this reaction can be detected during the reaction using fluorescent probes when incubations are conducted in a fluorimeter. Alternatively, products can be detected after amplification using a disposable amplicon containment device that contains an embedded lateral flow strip. Copyright © 2013 John Wiley & Sons, Inc.

  15. The membrane may be an important factor in browning of fresh-cut pear.

    Science.gov (United States)

    Li, Zhenghong; Zhang, Yuxing; Ge, Huibo

    2017-09-01

    Surface browning is an important cause of deterioration of fresh-cut fruit during postharvest handling. In this paper, four pear cultivars with different extents of natural browning were selected to analyse the factors involved in browning. The main results are as follows: the lipoxygenase (LOX) activity of 'Mantianhong' and 'Yali' pears was higher accompanied by a stronger degree of browning, while the LOX activity in 'Xueqing' and 'Xinli 7' pears was lower, with less browning. A higher unsaturated fatty acid ratio of pear resulted in reduced browning. The cell membranes disappeared 30min after being cut in 'Mantianhong' pear, which browns easily; however, the cell membranes were still intact 30min after being cut in 'Xueqing' pear, which does not brown easily. Therefore, it can be assumed that the stability of the cell membrane plays an important role in inhibiting browning of fresh-cut pears. Copyright © 2017. Published by Elsevier Ltd.

  16. Highly sensitive and label-free electrochemical detection of microRNAs based on triple signal amplification of multifunctional gold nanoparticles, enzymes and redox-cycling reaction.

    Science.gov (United States)

    Liu, Lin; Xia, Ning; Liu, Huiping; Kang, Xiaojing; Liu, Xiaoshuan; Xue, Chan; He, Xiaoling

    2014-03-15

    MicroRNAs (miRNAs) are believed to be important for cancer diagnosis and prognosis, serving as reliable molecular biomarkers. In this work, we presented a label-free and highly sensitive electrochemical genosensor for miRNAs detection with the triple signal amplification of gold nanoparticles (AuNPs), alkaline phosphatase (ALP) and p-aminophenol (p-AP) redox cycling. The label-free strategy is based on the difference in the structures of RNA and DNA. Specifically, miRNAs were first captured by the pre-immobilized DNA probes on a gold electrode. Next, the cis-diol group of ribose sugar at the end of the miRNAs chain allowed 3-aminophenylboronic acid (APBA)/biotin-modified multifunctional AuNPs (denoted as APBA-biotin-AuNPs) to be attached through the formation of a boronate ester covalent bond, which facilitated the capture of streptavidin-conjugated alkaline phosphatase (SA-ALP) via the biotin-streptavidin interaction. After the addition of the 4-aminophenylphosphate (p-APP) substrate, the enzymatic conversion from p-APP to p-AP occurred. The resulting p-AP could be cycled by a chemical reducing reagent after its electro-oxidization on the electrode (known as p-AP redox cycling), thus enabling an increase in the anodic current. As a result, the current increased linearly with the miRNAs concentration over a range of 10 fM-5 pM, and a detection limit of 3 fM was achieved. We believe that this work will be valuable for the design of new types of label-free and sensitive electrochemical biosensors. © 2013 Published by Elsevier B.V.

  17. Effect of fertilization methods on growth of pear trees, yielding and fruit quality

    Directory of Open Access Journals (Sweden)

    Tomasz Lipa

    2013-04-01

    Full Text Available The experiment was carried out in the commercial orchard near Lublin on five-year old pear trees of two cultivars: ‘Conference’and ‘Lukasowka’, planted on Quince MA. The objective of the study was evaluated the reaction of pear on the method of application of fertilizers. In the early spring the surface broadcasting of fertilizers was used (N – 71,5 kg, P 2O5 – 33,0 kg, K 2 O – 114,0 kg and from the May to the middle of August the fertigation was applied (N – 76,4 kg, P 2O5 – 49,5 kg, K 2 O – 84,2 kg. Method of fertilizer’s applications had no significant effect on the growth of evaluated trees. There were no differences in quantity of yield in dependence on the method of fertilization. In the case of cv. ‘Lukasowka’the beneficial influence of the way of fertilization on morphology of fruits (their diameter, height and mass was stated. Such an effect was not observed in the case of the ‘Conference’.

  18. Two pear glutathione S-transferases genes are regulated during fruit development and involved in response to salicylic acid, auxin, and glucose signaling.

    Directory of Open Access Journals (Sweden)

    Hai-Yan Shi

    Full Text Available Two genes encoding putative glutathione S-transferase proteins were isolated from pear (Pyrus pyrifolia and designated PpGST1 and PpGST2. The deduced PpGST1 and PpGST2 proteins contain conserved Glutathione S-transferase N-terminal domain (GST_N and Glutathione S-transferase, C-terminal domain (GST_C. Using PCR amplification technique, the genomic clones corresponding to PpGST1 and PpGST2 were isolated and shown to contain two introns and a singal intron respectively with typical GT/AG boundaries defining the splice junctions. Phylogenetic analysis clearly demonstrated that PpGST1 belonged to Phi class of GST superfamilies and had high homology with apple MdGST, while PpGST2 was classified into the Tau class of GST superfamilies. The expression of PpGST1 and PpGST2 genes was developmentally regulated in fruit. Further study demonstrated that PpGST1 and PpGST2 expression was remarkably induced by glucose, salicylic acid (SA and indole-3-aceticacid (IAA treatments in pear fruit, and in diseased fruit. These data suggested that PpGST1 and PpGST2 might be involved in response to sugar, SA, and IAA signaling during fruit development of pear.

  19. Some chemical and physico-mechanical properties of pear cultivars

    African Journals Online (AJOL)

    STORAGESEVER

    2009-02-18

    Feb 18, 2009 ... carried out at the Biological Material Laboratory in Agricultural. Machinery Department and Fruit Science Laboratory in Horticulture. Department of Ataturk University, Erzurum, Turkey. Chemical analysis. All chemical properties of the pear cultivars were investigated on randomly selected fifty fruit samples.

  20. Microorganisms associated with the spoilage of avocado pear ...

    African Journals Online (AJOL)

    The microorganisms associated with the spoilage of Avocado pear, Persea americana fruits, purchased fresh from various markets in Benin City were investigated. The pour plate method was used for the isolation. A total of nine species of microorganisms were isolated and identified in this study. They comprise of seven ...

  1. Potential causes of the pear thrips outbreak in sugar maple

    Science.gov (United States)

    Jack C. Schultz

    1991-01-01

    No one knows what caused the 1988 outbreak of pear thrips, Taeniothrips inconsequens (Uzel), in sugar maple, Acer saccharum Marsh., in the northeastern United States. As an entomologist and ecologist who knows even less about this insect than most of the authors of this volume, I cannot presume to understand the causes of this...

  2. Response of cactus pear genotypes to different crop densities

    Directory of Open Access Journals (Sweden)

    Lucas Aroaldo Dantas Cavalcante

    2014-09-01

    Full Text Available Considering the importance of cactus pear as an alimentary alternative for the herd of cattle of the Brazilian semiarid region and the effect of crop spacing among plants, this study aimed to evaluate the morphometry, yield and chemical-bromatological composition of cactus pear genotypes, under different cropping densities. The experimental design was completely randomized blocks, in a 3x4 factorial scheme. Treatments consisted of a combination of three cactus pear genotypes (Gigante, Redonda and Miúda and four cropping densities (10,000 plants ha-1; 20,000 plants ha-1; 40,000 plants ha-1; and 80,000 plants ha-1, with three replications. The cactus pear genotypes reacted differently, regarding morphometry, yield and chemical-bromatological composition, and, regardless of the species, the denser planting increased yield (tons ha-1. The Miúda palm presented the highest dry matter yield and consequently the greater accumulation of total digestible nutrients, raw protein and water per hectare, as well as the highest in vitro dry matter digestibility.

  3. Evaluation of six nucleic acid amplification tests used for diagnosis of Neisseria gonorrhoeae in Russia compared with an international strictly validated real-time porA pseudogene polymerase chain reaction.

    Science.gov (United States)

    Shipitsyna, E; Zolotoverkhaya, E; Hjelmevoll, S O; Maximova, A; Savicheva, A; Sokolovsky, E; Skogen, V; Domeika, M; Unemo, M

    2009-11-01

    In Russia, laboratory diagnosis of gonorrhoea has been mainly based on microscopy only and, in some settings, relatively rare suboptimal culturing. In recent years, Russian developed and manufactured nucleic acid amplification tests (NAAT) have been implemented for routine diagnosis of Neisseria gonorrhoeae. However, these NAATs have never been validated to any international well-recognized diagnostic NAAT. This study aims to evaluate the performance characteristics of six Russian NAATs for N. gonorrhoeae diagnostics. In total, 496 symptomatic patients were included. Five polymerase chain reaction (PCR) assays and one real-time nucleic acid sequence based amplification (NASBA) assay, developed by three Russian companies, were evaluated on urogenital samples, i.e. cervical and first voided urine (FVU) samples from females (n = 319), urethral and FVU samples from males (n = 127), and extragenital samples, i.e. rectal and pharyngeal samples, from 50 additional female patients with suspicion of gonorrhoea. As reference method, an international strictly validated real-time porA pseudogene PCR was applied. The prevalence of N. gonorrhoeae was 2.7% and 16% among the patients providing urogenital and extragenital samples, respectively. The Russian NAATs and the reference method displayed high level of concordance (99.4-100%). The sensitivities, specificities, positive predictive values and negative predictive values of the Russian tests in different specimens were 66.7-100%, 100%, 100%, and 99.4-100%, respectively. Russian N. gonorrhoeae diagnostic NAATs comprise relatively good performance characteristics. However, larger studies are crucial and, beneficially, the Russian assays should also be evaluated to other international highly sensitive and specific, and ideally Food and Drug Administration approved, NAATs such as Aptima Combo 2 (Gen-Probe).

  4. 78 FR 24033 - Pears Grown in Oregon and Washington; Modification of the Assessment Rate for Fresh Pears

    Science.gov (United States)

    2013-04-24

    ... March 2012 by the National Agricultural Statistics Service, the total 2011 farm-gate value of all pears...: 7 U.S.C. 601-674. 0 2. In Sec. 927.236, the introductory text and paragraphs (a) and (b) are revised...

  5. 78 FR 34 - Pears Grown in Oregon and Washington; Modification of the Assessment Rate for Fresh Pears

    Science.gov (United States)

    2013-01-02

    ... Agricultural Statistics Service, the total 2011 farm-gate value of all pears grown in Oregon and Washington is... follows: Authority: 7 U.S.C. 601-674. 0 2. In Sec. 927.236, the introductory text and paragraphs (a) and...

  6. Enzyme-free electrochemical detection of microRNA-21 using immobilized hairpin probes and a target-triggered hybridization chain reaction amplification strategy

    International Nuclear Information System (INIS)

    Liu, Hongying; Bei, Xiaoqiong; Xia, Qiuting; Fu, Yan; Zhang, Shi; Liu, Maochuan; Fan, Kai; Zhang, Mingzhen; Yang, Yong

    2016-01-01

    We describe a sensitive enzyme-free bioassay for the determination of microRNA-21. It is based on a combination of target-triggered hybridization chain reaction, tagging with CdTe quantum dots (QDs), and anodic stripping voltammetry. Firstly, a thiolated capture hairpin probe SH-HP1 was immobilized on the surface of a gold electrode. HP1 unfolds in the presence of microRNA-21. If hairpin probe 2 (HP2) is present, a HP1-HP2 complex will be formed which possesses an exposed stem of HP2, and microRNA is released in parallel. The released microRNA-21 triggers a hybridization chain reaction and this leads to form an exposed DNA segment of HP2 and cycle use microRNA-21. With the aid of assistant DNA A1 and A2, the exposed DNA segment of HP2 progressed to a long double strand. The strand is rich in CdTe QDs with the help of QDs-A1. Then, the attached QDs were dissolved with HNO 3 to give dissolved Cd(II) ions. Finally, the corresponding electrochemical current response of Cd(II) is monitored by anodic stripping voltammetry and used to quantify the concentration of microRNA-21. More microRNA-21 participated in this reaction increases the number of CdTe QDs, which results in increased electrochemical current. Thus, an ultrasensitive detection of microRNA-21 is accomplished by anodic stripping voltammetry. This gene assay displays a detection limit as low as 33 aM. It can discriminate between complementary DNA sequence and single-base mismatched DNA, indicating its high specificity. (author)

  7. Identification of Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) by using polymerase chain reaction amplification and restriction analysis of the mitochondrial cytochrome b gene.

    Science.gov (United States)

    Carrera, E; García, T; Céspedes, A; González, I; Sanz, B; Hernández, P E; Martín, R

    1998-04-01

    Restriction site analysis of polymerase chain reaction (PCR) products from a conserved region of the cytochrome b gene has been used for the identification of fresh and smoked samples of Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss). Digestion of the 359-bp PCR product with the endonucleases EcoRV and TaqI yielded specific banding patterns for salmon and trout. This genetic marker can be very useful for detecting fraudulent substitution of the cheaper smoked trout for the more expensive smoked salmon.

  8. Transcriptomic analysis of ‘Suli’ pear (Pyrus pyrifolia white pear group buds during the dormancy by RNA-Seq

    Directory of Open Access Journals (Sweden)

    Liu Guoqin

    2012-12-01

    Full Text Available Abstract Background Bud dormancy is a critical developmental process that allows perennial plants to survive unfavorable environmental conditions. Pear is one of the most important deciduous fruit trees in the world, but the mechanisms regulating bud dormancy in this species are unknown. Because genomic information for pear is currently unavailable, transcriptome and digital gene expression data for this species would be valuable resources to better understand the molecular and biological mechanisms regulating its bud dormancy. Results We performed de novo transcriptome assembly and digital gene expression (DGE profiling analyses of ‘Suli’ pear (Pyrus pyrifolia white pear group using the Illumina RNA-seq system. RNA-Seq generated approximately 100 M high-quality reads that were assembled into 69,393 unigenes (mean length = 853 bp, including 14,531 clusters and 34,194 singletons. A total of 51,448 (74.1% unigenes were annotated using public protein databases with a cut-off E-value above 10-5. We mainly compared gene expression levels at four time-points during bud dormancy. Between Nov. 15 and Dec. 15, Dec. 15 and Jan. 15, and Jan. 15 and Feb. 15, 1,978, 1,024, and 3,468 genes were differentially expressed, respectively. Hierarchical clustering analysis arranged 190 significantly differentially-expressed genes into seven groups. Seven genes were randomly selected to confirm their expression levels using quantitative real-time PCR. Conclusions The new transcriptomes offer comprehensive sequence and DGE profiling data for a dynamic view of transcriptomic variation during bud dormancy in pear. These data provided a basis for future studies of metabolism during bud dormancy in non-model but economically-important perennial species.

  9. Reaction

    African Journals Online (AJOL)

    abp

    19 oct. 2017 ... Reaction to Mohamed Said Nakhli et al. concerning the article: "When the axillary block remains the only alternative in a 5 year old child". .... Bertini L1, Savoia G, De Nicola A, Ivani G, Gravino E, Albani A et al ... 2010;7(2):101-.

  10. Detection and quantitative characterization of artificial extra peaks following polymerase chain reaction amplification of 14 short tandem repeat systems used in forensic investigations

    DEFF Research Database (Denmark)

    Meldgaard, Michael; Morling, N

    1997-01-01

    Detection on automated DNA sequencers of polymerase chain reaction (PCR) products of tetra- and penta-nucleotide short tandem repeat (STR) loci frequently reveals one or more extra peaks along with the true, major allele peak. The most frequent extra peak pattern is a single smaller peak which...... is one repeat unit shorter than the true allele peak. The existence of such artificial peaks is of special importance when the methods are used for forensic investigations because the artificial extra peaks may simulate true alleles when samples containing mixtures of DNA from different individuals...... are analyzed. We have investigated the relative levels of formation of extra peaks in 14 STR marker systems. We found that not only the parameters of the PCR but also factors determining the stringency during the post-PCR and pre-electrophoresis handling of samples were of importance for the formation of extra...

  11. [Investigation of RNA viral genome amplification by multiple displacement amplification technique].

    Science.gov (United States)

    Pang, Zheng; Li, Jian-Dong; Li, Chuan; Liang, Mi-Fang; Li, De-Xin

    2013-06-01

    In order to facilitate the detection of newly emerging or rare viral infectious diseases, a negative-strand RNA virus-severe fever with thrombocytopenia syndrome bunyavirus, and a positive-strand RNA virus-dengue virus, were used to investigate RNA viral genome unspecific amplification by multiple displacement amplification technique from clinical samples. Series of 10-fold diluted purified viral RNA were utilized as analog samples with different pathogen loads, after a series of reactions were sequentially processed, single-strand cDNA, double-strand cDNA, double-strand cDNA treated with ligation without or with supplemental RNA were generated, then a Phi29 DNA polymerase depended isothermal amplification was employed, and finally the target gene copies were detected by real time PCR assays to evaluate the amplification efficiencies of various methods. The results showed that multiple displacement amplification effects of single-strand or double-strand cDNA templates were limited, while the fold increases of double-strand cDNA templates treated with ligation could be up to 6 X 10(3), even 2 X 10(5) when supplemental RNA existed, and better results were obtained when viral RNA loads were lower. A RNA viral genome amplification system using multiple displacement amplification technique was established in this study and effective amplification of RNA viral genome with low load was achieved, which could provide a tool to synthesize adequate viral genome for multiplex pathogens detection.

  12. FIRE BLIGHT SUSCEPTIBILITY OF SOME PEAR VARIETIES (ERWINIA AMYLOVORA, BURILL

    Directory of Open Access Journals (Sweden)

    Zsolt Jakab- Ilyefalv

    2012-01-01

    Full Text Available At Bistriţa Fruit Research and Development Station, in a pear collection, planted with 44 varieties, there has been studied the susceptibility to fire blight (Erwinia amylovora,Burill. During the vegetation period, phytosanitary treatments to control the disease have been applied using copper based products (copper sulfate, Champion, Funguran, Kocide. Erwinia amylovora infections have been influenced by the rainfall registered in April (70.9 mm and May (104.7 mm and the temperature fluctuations in April-May. The field observations on Erwinia amylvora attack demonstrate that the pear varieties have a different susceptibility to this dangerous bacterium. Evaluation of attack level in the pear collection was done using an evaluation scale with 9 scores using a reference resistance scale for : ‘Highly resistant’ , ‘Moderately resistant’, ‘Susceptible’, ‘Very susceptible’ cultivars . The most sensitive pear varieties in the collection were: ‘De Noiembrie’, ‘Abatele Fetel’, ’Daciana’, ’Triumf’, ’Williams Boway’, ’Margareta Marillat’, ’Beauty Tomme’, ‘Williams rosu’, ’Aromata de Bistrita’, ’Jeanne d`Arc’, ’Aramiu de Somes’, ’Belle des Arbres’, ’Zorka’ representing 13.64% of the total pear varieties. In several cultivars there have been observed increased symptoms, a very high susceptibility of infections leading to complete scorching of trees: Jeanne d`Arc’, Williams rosu’, ‘Triumf’, ‘Aromata de Bistrita’, ‘Zorka’. Strong attack symtoms were observed at the pear cultivars ‘Cure’, ‘Euras, ’Ciuda’ ‘Highland’, ‘Precoce Morettini’, ’Monica’, ’Cadillac’, ’Juliana’, ’Somesan’, ’Beurré Hardenpont’ these cultivars being susceptible to Erwinia amylovora , representing 40.91 % of total genotypes . Cultivars ‘Untoasa Geoagiu’, ‘Beurre Hardy’, ‘Trivale’ manifested a certain tolerance to Erwinia amylovora , no attack symptoms being

  13. Environment-Friendly Control of Pear Scab and Rust Using Lime Sulfur

    Directory of Open Access Journals (Sweden)

    Ju Hoon Cha

    2018-03-01

    Full Text Available Pear scab and rust are the major diseases causing severe epidemics in organic cultivation of pear trees. Incidences of pear scab and rust were compared in organically managed plots and conventionally managed plots to obtain optimum application schedule of environment-friendly control agents in organically managed plots. Organically cultural practice with 10 time-applications of lime sulfur and Bordeaux mixture showed higher than 40% of control efficacies of pear scab and rust compared to conventionally cultural practice. Organically cultural practice with 8 time-applications of lime sulfur considering weather condition showed higher than 30% of control efficacies of pear scab compared to conventionally cultural practice. The results suggest that proper application of environment-friendly control agents such as lime sulfur considering weather condition will enable effective control of the major diseases for organic cultivation of pear.

  14. Selective amplification of T-cell receptor variable region species is demonstrable but not essential in early lesions of psoriasis vulgaris: analysis by anchored polymerase chain reaction and hypervariable region size spectratyping.

    Science.gov (United States)

    Vekony, M A; Holder, J E; Lee, A J; Horrocks, C; Eperon, I C; Camp, R D

    1997-07-01

    Several groups have investigated the role of T cells in the pathogenesis of psoriasis by determination of T-cell receptor (TCR) B-chain variable (V) region usage, both in chronic plaque (psoriasis vulgaris) and guttate forms, with various results. Because there are no data on TCR expression in early psoriasis vulgaris, when specific cellular immune events may be expected to be most pronounced, we have analyzed early lesions (less than 3 wk old) of ten patients, with highly reproducible results. We have developed a highly controlled anchored polymerase chain reaction (PCR) method in which TCR beta chain species are all amplified with the same primer pair and products are quantified by dot blot hybridization with BV family-specific oligonucleotide probes. Overexpression of certain TCR BV genes was observed in the majority of lesional biopsies, but in samples in which the expanded BV family formed more than 10% of total lesional BV (half of the samples analyzed), BV2 and BV6 predominated. The consistency of overexpression of these BV species between patients was much less than in previous studies of TCRBV usage in established chronic plaque psoriasis lesions. Complementarity-determining region 3 (CDR3) size spectratyping demonstrated evidence for selective clonal T cell accumulation in less than half of the lesional samples showing BV expansion. These results indicate that selective amplification of TCRBV species occurs in early psoriasis vulgaris but is not essential to the pathogenic process and may be more important in the maintenance or expansion of chronic lesions.

  15. Studies on Perspectives and Marketing Strategies of Pear Farming in Japan

    OpenAIRE

    竹内, 幹雄; 佐藤, 豊信; 目瀬, 守男

    1992-01-01

    As the economic situations surrounding pear farms are changing rapidly, a new management is required to cope with them. This paper is to study the past and present conditions of pear marketing in Tottori and to suggest a future strategy. At present pears have been marketed primarily through individual cooperative with some coordinations and supports by Tottori Fruit Growers Cooperative Association. But recently the consummers needs have been changing for higher quality fruit, the improvemen...

  16. Industrialization Development of Korla Fragrant Pear in Bayingolin Mongol Autonomous Prefecture, China

    OpenAIRE

    Ma, Qiong; Li, Ying

    2010-01-01

    Based on the introduction of the natural and geographic conditions in Bayingolin Mongol Autonomous Prefecture in Xinjiang(Bazhou), development status of Korla Fragrant Pear is introduced from the two aspects of the production status and the storage and processing status of Korla Fragrant Pear. Among them, production status of Korla Fragrant Pear is analyzed from the aspects of the rapid growth of planting area and the stable growth of output. And the storage and processing status of Korla Fra...

  17. Phytotoxic effects of phenylmercuric compounds upon certain pear varieties

    Energy Technology Data Exchange (ETDEWEB)

    Kirby, A H.M.; Bennett, M

    1963-01-01

    In trials at East Malling Research Station, extending over the years 1952 to 1959, chlorotic spotting, associated with loss of chlorophyll, and distortion of leaves occurred following post-blossom application to four pear varieties of phenylmercuric salts at 0.003% elemental mercury concentration. In the light of rainfall and temperature conditions recorded on the days following each post-blossom application in each year, crop loss must be expected following post-blossom application of sufficient organomercurial for scab control in any season on Conference and Doyenne du Comice; on Conference, reduction of fruit-bud formation may also occur. Although pear leaves are obviously damaged by phenylmercuric salts, it is likely that the effects on cropping are largely due to direct absorption and/or translocation of some form of mercury into fruit-buds and fruitlets.

  18. An enzymatic signal amplification system for calorimetric studies of cellobiohydrolases

    DEFF Research Database (Denmark)

    Murphy, Leigh; Baumann, Martin Johannes; Borch, Kim

    2010-01-01

    amplification method has been developed to measure even slow hydrolytically active enzymes such as cellobiohydrolases. This method is explained in detail for the amplification of the heat signal by more than 130 times by using glucose oxidase and catalase. The kinetics of this complex coupled reaction system...

  19. Antioxidant and Anticlastogenic Capacity of Prickly Pear Juice

    Directory of Open Access Journals (Sweden)

    Alejandra Hernández-Ceruelos

    2013-10-01

    Full Text Available Plants belonging to the genus Opuntia spp. are the most abundant of the Cactaceae family, grown throughout America and the Mediterranean central area. Its fruit, known as cactus pear or prickly pear, is an oval berry grouped in different colors. Some studies have shown its antioxidant activities which may help in preventing chronic pathologies such as diabetes. The purpose of the study was to evaluate the antioxidant capacity of three varieties of prickly pear juice (red-purple, white-green and yellow-orange in five different concentrations (100, 250, 500, 750, and 1000 mg/mL by DPPH (1,1-diphenyl-2-picrylhydrazyl radical colorimetric method, selecting the best variety to determine its anticlastogenic potential against methyl methanesulfonate (MMS. The results indicate that the highest antioxidant was found in the juice of the prickly pear red-purple variety (PPRP, in all concentrations. Its anticlastogenic potential was therefore evaluated with a micronucleus assay. The experiment was run over two weeks. A negative control was included along with a positive control with MMS (40 mg/kg, a group of mice treated with PPRP (25 mL/kg, and three groups with PPRP (in doses of 25, 16.5 and 8.3 mL/kg plus the mutagen. The PPRP was administered daily by oral gavage and the MMS was injected intraperitoneally five days prior to the end of the experiment. Blood samples were obtained at 0, 24, 48, 72 and 96 h in order to determine the frequency of micronucleated polychromatic erythrocytes (MNPE. The results indicated that PPRP is not a genotoxic agent, on the contrary, it may reduce the number of MNPE. In this regard, the PPRP showed an anticlastogenic effect directly proportional to its concentrations. Thus, the highest protection was obtained with a concentration of 25 mL/kg after 48 h of treatment.

  20. Bioecology of pear thrips: distribution in forest soils

    Science.gov (United States)

    Margaret Skinner; Bruce L. Parker

    1991-01-01

    The vertical and horizontal distribution of pear thrips in Vermont sugar maple forest soils was investigated. In the fall, about 86% of the thrips were found in the upper 10 cm of soil, though a few were found as deep as 20 cm. No thrips were found in the leaf litter. Soil sampling tools to determine thrips populations within an entire forest were tested and a standard...

  1. REPEATABILITY OF FRUIT QUALITY TRAITS OF CACTUS PEAR

    Directory of Open Access Journals (Sweden)

    VALTÂNIA XAVIER NUNES

    2017-12-01

    Full Text Available ABSTRACT Repeatability analysis has been used to study traits in several crops, assisting in the definition of the minimum number needed to evaluate genotypes more efficiently and with less time and resource consumption. So far, however, no repeatability studies on cactus pear have been found in the literature. The objective of this study was to determine the coefficient of repeatability for cactus pear fruits traits and the minimum number of evaluations (fruit that can provide acceptable accuracy for the prediction of the true value. The experiment was conducted at the Federal Institute of Bahia/Campus Guanambi, with 150 fruits collected from three municipalities in the state of Bahia. The coefficients of repeatability were estimated by the methods of analysis of variance, principal components based on the covariance (PCCV and correlation (PCC matrices, and structural analysis based on the correlation matrix (SA. The analysis of variance showed that, except for fruit diameter, the effect of the production site (municipality was significant for all traits evaluated. The PCCV method was proven the most suitable for studying the repeatability of quality traits of cactus pear fruits. Seven fruits were required to determine, with 90% confidence, the traits length, diameter, fruit firmness, skin thickness, number of seeds, fruit mass, bark mass, pulp mass, pH, titratable acidity, soluble solids, SS/AT ratio, and pulp yield.

  2. ALTERNATIVE FOR REDUCING PHYSIOLOGICAL DISORDERS IN ‘BARTLETT’ PEARS

    Directory of Open Access Journals (Sweden)

    MOISES ZUCOLOTO

    2016-01-01

    Full Text Available ABSTRACT ‘Bartlett’ pears from different harvest dates were assessed regarding to cold storage potential and reduction of physiological disorder incidence. Three harvests, the first (HD1, second (HD2, and third (HD3, were carried out at weekly intervals. The pears were assessed after the harvest, with no exposition to the temperature conditioning, after 20, 40, 60, 80, 100 and 120 days at 0 ± 1 °C and 90 ± 5% RH and after three and six days at room temperature (20 ± 1 °C. Fruit from the early harvest (HD1 showed the smallest incidence of physiological disorder during both cold and room temperature storage. The disorder symptoms became apparent in HD1 fruit after 20 days at cold storage followed by three days at 20 °C, whereas HD2 and HD3 fruit showed the symptoms before being kept in a cold room. ‘Bartlett’ pears harvested at 70.75 N flesh firmness can be stored at 0 ± 1 °C for up to 40 days and preferably commercialized within three days, when they reach the firmness for eating. The extension of cold storage as well as the trade period can result in higher physiological disorder incidence and loss of sensorial quality.

  3. Loop-mediated isothermal amplification (LAMP) based detection of ...

    African Journals Online (AJOL)

    SAM

    2014-05-07

    May 7, 2014 ... 2 months for growing in a culture. Therefore, to control .... The LAMP reaction is carried out in a 25 µL reaction mixture containing ..... J. Fish Dis. 32(6):491-497. Goto M, Honda E, Ogura A, Nomoto A, Hanaki K (2009). Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy ...

  4. Strategies to control tree vigour and optimise fruit production in 'Conference' pears

    NARCIS (Netherlands)

    Maas, F.M.

    2008-01-01

    The ban on the use of chlormequat (CCC) in pear orchards in 2001 forced Dutch pear growers to look for alternative methods to control tree vigour and stimulate flower bud development and fruit production. Root pruning and trunk notching have become the major growth retarding methods. In addition to

  5. Classification of Korean native pear based on a standard set of microsatellite loci

    Science.gov (United States)

    After analysis with 12 microsatellite loci previously selected for use by ECPGR, 34 Korean pear accessions were classified into 3 groups. Group I included 4 Korean pea pears and one accession of P. calleryana. Group II contained 12 cultivars including 7 accessions presumed to belong to P. pyrifolia....

  6. Adulteration of apple with pear juice: emphasis on major carbohydrates, proline, and arbutin.

    Science.gov (United States)

    Thavarajah, Pushparajah; Low, Nicholas H

    2006-06-28

    Detection of juice-to-juice adulteration based on chemical composition studies is a common method used by government regulatory agencies and food companies. This study investigated the use of major carbohydrate (fructose, glucose and sucrose), polyol (sorbitol), proline, and phenolic profiles as indicators of pear adulteration of apple juice (PAAJ). For this work, a total of 105 authentic apple juice samples from 13 countries and 27 authentic pear juice samples from 5 countries were analyzed. Because the major carbohydrate ranges for these juices showed significant overlap their use as markers for PAAJ detection would be very limited. It was found that sorbitol and proline means for apple and pear were significantly different; however, their broad natural ranges would afford PAAJ at levels up to 30% without detection. In addition, careful selection of the pear juice used as the adulterant would further limit the usefulness of these markers for PAAJ detection. Arbutin was conclusively identified as a marker for pear juice on the basis of its presence in all 27 authentic pear samples and its absence (apple juice samples analyzed in this study. The application of the developed HPLC-PDA method for arbutin analysis to detect PAAJ at levels as low as 2% (v/v) was demonstrated. A confirmation method for the presence of arbutin in pure pear juice and apple adulterated with pear juice was introduced on the basis of the hydrolysis of arbutin to hydroquinone employing beta-glucosidase, with reactant and product monitoring by HPLC-PDA.

  7. Application of exogenous ethylene inhibits postharvest peel browning of ‘Huangguan’ pear

    Science.gov (United States)

    Peel browning disorder has an enormous impact on the exterior quality of ‘Huangguan’ pear whereas the underlying mechanism is still unclear. In this study, the effect of exogenous ethylene on peel browning of pear fruits stored at 0' was evaluated. Results showed that ethylene effectively inhibited ...

  8. 76 FR 27848 - Pears Grown in Oregon and Washington; Amendment To Allow Additional Exemptions

    Science.gov (United States)

    2011-05-13

    ... marketing order for Oregon-Washington pears that provides for the sale of fresh pears directly to consumers...' markets. These consumer-direct sales are exempt from the marketing order's assessment, reporting, handling... DEPARTMENT OF AGRICULTURE Agricultural Marketing Service 7 CFR Part 927 [Doc. No. AMS-FV-10-0072...

  9. The relationship between measures of tree vigor and pear thrips damage in sugar maple

    Science.gov (United States)

    Gretchen Smith; Christina M. Petersen; Roy Van Driesche; Charles Burnham

    1991-01-01

    In this presentation I will address three points associated with pear thrips damage and sugar maple. First, I will describe the impact of pear thrips on sugar maple in Massachusetts, in both the sugarbush and the natural forest stand, based on root starch assays that were completed this fall (1988). Secondly, I will discuss the relationship between tree health and...

  10. The life cycle of pear thrips, Taeniothrips inconsequens (Uzel) in Vermont

    Science.gov (United States)

    Margaret Skinner; Bruce L. Parker; Sandra H. Wilmot

    1991-01-01

    Life history information has been collected for pear thrips in orchard habitats in California and British Columbia (Cameron & Treherne 1918, Bailey 1944). However such information is not available for this insect in a northern hardwood forest ecosystem in the eastern United States. Research is currently underway at the University of Vermont to determine the pear...

  11. 75 FR 77563 - Nectarines, Pears, and Peaches Grown in California; Continuance Referenda

    Science.gov (United States)

    2010-12-13

    ... that time, the pear industry has been regulated by a State marketing order. If the results of the pear... DEPARTMENT OF AGRICULTURE Agricultural Marketing Service 7 CFR Parts 916 and 917 [Doc. No. AMS-FV... AGENCY: Agricultural Marketing Service, USDA. ACTION: Referenda order. SUMMARY: This document directs...

  12. Draft genome sequence of Xylella fastidiosa pear leaf scorch strain in Taiwan

    Science.gov (United States)

    The draft genome sequence of Xylella fastidiosa pear leaf scorch strain (PLS229) isolated from pear cultivar Hengshan (Pyrus pyrifolia) in Taiwan is reported. The bacterium has a genome size of 2,733,013 bp with a G+C content of 53.1%. The PLS229 strain genome was annotated to have 3,259 open readin...

  13. 7 CFR 319.56-22 - Apples and pears from certain countries in Europe.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 5 2010-01-01 2010-01-01 false Apples and pears from certain countries in Europe. 319.56-22 Section 319.56-22 Agriculture Regulations of the Department of Agriculture (Continued) ANIMAL... Vegetables § 319.56-22 Apples and pears from certain countries in Europe. (a) Importations allowed. The...

  14. Constitutive and herbivore-induced volatiles in pear, alder and hawthorn trees

    NARCIS (Netherlands)

    Scutareanu, P.; Bruin, de J.; Posthumus, M.A.; Drukker, B.

    2003-01-01

    Qualitative and quantitative differences among pear cultivars were found in constitutive and Cacopsylla-induced volatiles, depending on experimental treatment of the trees (i.e., uninfested and partly or completely infested by psyllids). Blend differences were also found between pear cultivars and

  15. Prickly pear induces upregulation of liver LDL binding in familial heterozygous hypercholesterolemia

    International Nuclear Information System (INIS)

    Palumbo, B.; Palumbo, R.; Efthimiou, Y.; Stamatopoulos, J.; Sinzinger, H.; Oguogho, A.; Budinsky, A.; Sinzinger, H.

    2003-01-01

    The hypoglycemic effect of prickly pear is well known by native local Indian population since a long time. Beside the beneficial effects on lipid metabolism, oxidation injury and platelet function has been claimed in experimental animals. We recently found an upregulation of apo-B/E receptor. We therefore examined 10 patients with isolated heterozygous familial hypercholesterolemia (FH) being enrolled in a dietary run-in phase of 6 weeks after dietary counselling and a further 6 weeks of prickly pear addition. Uptake of autologous 123 I-radiolabeled LDL was determined at entry as well as after 6 weeks of daily prickly pear ingestion. We found a significant (p 176.4 mg/dl; p 123 I-LDL binding by prickly pear in FH-patients in vivo and indicate that prickly pear exerts a significant hypolipidemic action via receptor upregulation. (author)

  16. Nutrient removal by apple, pear and cherry nursery trees

    Directory of Open Access Journals (Sweden)

    Giovambattista Sorrenti

    2017-06-01

    Full Text Available Given that nursery is a peculiar environment, the amount of nutrients removed by nursery trees represents a fundamental acquisition to optimise fertilisation strategies, with economic and environmental implications. In this context, we determined nutrient removal by apple, pear and cherry nursery trees at the end of the nursery growing cycle. We randomly removed 5 leafless apple (Golden Delicious/EMLA M9; density of 30,000 trees ha–1, pear (Santa Maria/Adams; density of 30,000 trees ha–1 and cherry (AlexTM/Gisela 6®; density of 40,000 trees ha–1 trees from a commercial nursery. Trees were divided into roots (below the root collar, rootstock (above-ground wood between root collar and grafting point and variety (1-year-old wood above the grafting point. For each organ we determined biomass, macro- (N, P, K, Ca, Mg, S, and micro- (Fe, Mn, Zn, Cu, and B nutrient concentration. Pear trees were the most developed (650 g (dw tree–1, equal to 1.75 and 2.78 folds than apple and cherry trees, respectively whereas, independently of the species, variety mostly contributed (>50% to the total tree biomass, followed by roots and then above-ground rootstock. However, the dry biomass and nutrient amount measured in rootstocks (including roots represent the cumulative amount of 2 and 3 seasons, for Gisela® 6 (tissue culture and pome fruit species (generated by mound layering, respectively. Macro and micronutrients were mostly concentrated in roots, followed by variety and rootstock, irrespective of the species. Independently of the tissue, macronutrients concentration hierarchy was N>Ca>K> P>Mg>S. Removed N by whole tree accounted for 6.58, 3.53 and 2.49 g tree–1 for pear, apple and cherry, respectively, corresponding to almost 200, 107 and 100 kg N ha–1, respectively. High amounts of K and Ca were used by pear (130-140 kg ha–1 and apple trees (~50 and 130 kg ha–1 of K and Ca, respectively, while ~25 kg K ha–1 and 55 kg Ca ha–1 were

  17. Gene amplification in carcinogenesis

    Directory of Open Access Journals (Sweden)

    Lucimari Bizari

    2006-01-01

    Full Text Available Gene amplification increases the number of genes in a genome and can give rise to karyotype abnormalities called double minutes (DM and homogeneously staining regions (HSR, both of which have been widely observed in human tumors but are also known to play a major role during embryonic development due to the fact that they are responsible for the programmed increase of gene expression. The etiology of gene amplification during carcinogenesis is not yet completely understood but can be considered a result of genetic instability. Gene amplification leads to an increase in protein expression and provides a selective advantage during cell growth. Oncogenes such as CCND1, c-MET, c-MYC, ERBB2, EGFR and MDM2 are amplified in human tumors and can be associated with increased expression of their respective proteins or not. In general, gene amplification is associated with more aggressive tumors, metastases, resistance to chemotherapy and a decrease in the period during which the patient stays free of the disease. This review discusses the major role of gene amplification in the progression of carcinomas, formation of genetic markers and as possible therapeutic targets for the development of drugs for the treatment of some types of tumors.

  18. Phytochemical study of prickly pear from southern Morocco

    Directory of Open Access Journals (Sweden)

    Z. Bouzoubaâ

    2016-06-01

    Full Text Available This work concerns the phytochemical study of the prickly pear pulp’s fruits of two opuntia cultivars; Achefri and Amouslem widely present in two regions of southern Morocco; Arbaa Sahel and Asgherkis that are different in their altitude and annual rainfall. The results of the phytochemical study show that the levels of antioxidants have a non-significant difference between the fruits of the two sites (comparing Amouslem and Achefri in the same site, on the one hand, for the differences due to the variety or cultivar, on the other hand between Amouslem and Achefri from the two sites to show the site effect.

  19. Social amplification of risk

    International Nuclear Information System (INIS)

    Kasperson, R.E.; Renn, O.; Slovic, P.; Kasperson, J.X.; Emani, S.

    1989-01-01

    The risks associated with radioactive and other hazardous waste disposal may be expected to interact with societal processes to enlarge or attenuate the consequences of risks and risk events. This article summarizes a conceptual framework that depicts the social amplification of risk. Using a data base of 128 hazard events that have occurred largely over the past ten years, the authors examine the role of physical consequences, media coverage, and public perceptions of risk in generating social and economic impacts. The analysis concludes that social amplification processes substantially shape the nature and magnitude of those impacts but also that such social amplification appears to be systematically related to characteristics of the risks and risk events

  20. Exponential isothermal amplification of nucleic acids and amplified assays for proteins, cells, and enzyme activities.

    Science.gov (United States)

    Reid, Michael S; Le, X Chris; Zhang, Hongquan

    2018-04-27

    Isothermal exponential amplification techniques, such as strand-displacement amplification (SDA), rolling circle amplification (RCA), loop-mediated isothermal amplification (LAMP), nucleic acid sequence-based amplification (NASBA), helicase-dependent amplification (HDA), and recombinase polymerase amplification (RPA), have great potential for on-site, point-of-care, and in-situ assay applications. These amplification techniques eliminate the need for temperature cycling required for polymerase chain reaction (PCR) while achieving comparable amplification yield. We highlight here recent advances in exponential amplification reaction (EXPAR) for the detection of nucleic acids, proteins, enzyme activities, cells, and metal ions. We discuss design strategies, enzyme reactions, detection techniques, and key features. Incorporation of fluorescence, colorimetric, chemiluminescence, Raman, and electrochemical approaches enables highly sensitive detection of a variety of targets. Remaining issues, such as undesirable background amplification resulting from non-specific template interactions, must be addressed to further improve isothermal and exponential amplification techniques. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Telomerase Repeated Amplification Protocol (TRAP).

    Science.gov (United States)

    Mender, Ilgen; Shay, Jerry W

    2015-11-20

    Telomeres are found at the end of eukaryotic linear chromosomes, and proteins that bind to telomeres protect DNA from being recognized as double-strand breaks thus preventing end-to-end fusions (Griffith et al. , 1999). However, due to the end replication problem and other factors such as oxidative damage, the limited life span of cultured cells (Hayflick limit) results in progressive shortening of these protective structures (Hayflick and Moorhead, 1961; Olovnikov, 1973). The ribonucleoprotein enzyme complex telomerase-consisting of a protein catalytic component hTERT and a functional RNA component hTR or hTERC - counteracts telomere shortening by adding telomeric repeats to the end of chromosomes in ~90% of primary human tumors and in some transiently proliferating stem-like cells (Shay and Wright, 1996; Shay and Wright, 2001). This results in continuous proliferation of cells which is a hallmark of cancer. Therefore, telomere biology has a central role in aging, cancer progression/metastasis as well as targeted cancer therapies. There are commonly used methods in telomere biology such as Telomere Restriction Fragment (TRF) (Mender and Shay, 2015b), Telomere Repeat Amplification Protocol (TRAP) and Telomere dysfunction Induced Foci (TIF) analysis (Mender and Shay, 2015a). In this detailed protocol we describe Telomere Repeat Amplification Protocol (TRAP). The TRAP assay is a popular method to determine telomerase activity in mammalian cells and tissue samples (Kim et al. , 1994). The TRAP assay includes three steps: extension, amplification, and detection of telomerase products. In the extension step, telomeric repeats are added to the telomerase substrate (which is actually a non telomeric oligonucleotide, TS) by telomerase. In the amplification step, the extension products are amplified by the polymerase chain reaction (PCR) using specific primers (TS upstream primer and ACX downstream primer) and in the detection step, the presence or absence of telomerase is

  2. Multi-chamber nucleic acid amplification and detection device

    Science.gov (United States)

    Dugan, Lawrence

    2017-10-25

    A nucleic acid amplification and detection device includes an amplification cartridge with a plurality of reaction chambers for containing an amplification reagent and a visual detection reagent, and a plurality of optically transparent view ports for viewing inside the reaction chambers. The cartridge also includes a sample receiving port which is adapted to receive a fluid sample and fluidically connected to distribute the fluid sample to the reaction chamber, and in one embodiment, a plunger is carried by the cartridge for occluding fluidic communication to the reaction chambers. The device also includes a heating apparatus having a heating element which is activated by controller to generate heat when a trigger event is detected. The heating apparatus includes a cartridge-mounting section which positioned a cartridge in thermal communication with the heating element so that visual changes to the contents of the reaction chambers are viewable through the view ports.

  3. Controlled Microwave Heating Accelerates Rolling Circle Amplification.

    Science.gov (United States)

    Yoshimura, Takeo; Suzuki, Takamasa; Mineki, Shigeru; Ohuchi, Shokichi

    2015-01-01

    Rolling circle amplification (RCA) generates single-stranded DNAs or RNA, and the diverse applications of this isothermal technique range from the sensitive detection of nucleic acids to analysis of single nucleotide polymorphisms. Microwave chemistry is widely applied to increase reaction rate as well as product yield and purity. The objectives of the present research were to apply microwave heating to RCA and indicate factors that contribute to the microwave selective heating effect. The microwave reaction temperature was strictly controlled using a microwave applicator optimized for enzymatic-scale reactions. Here, we showed that microwave-assisted RCA reactions catalyzed by either of the four thermostable DNA polymerases were accelerated over 4-folds compared with conventional RCA. Furthermore, the temperatures of the individual buffer components were specifically influenced by microwave heating. We concluded that microwave heating accelerated isothermal RCA of DNA because of the differential heating mechanisms of microwaves on the temperatures of reaction components, although the overall reaction temperatures were the same.

  4. Biomaterials in light amplification

    Science.gov (United States)

    Mysliwiec, Jaroslaw; Cyprych, Konrad; Sznitko, Lech; Miniewicz, Andrzej

    2017-03-01

    Biologically produced or inspired materials can serve as optical gain media, i.e. they can exhibit the phenomenon of light amplification. Some of these materials, under suitable dye-doping and optical pumping conditions, show lasing phenomena. The emerging branch of research focused on obtaining lasing action in highly disordered and highly light scattering materials, i.e. research on random lasing, is perfectly suited for biological materials. The use of biomaterials in light amplification has been extensively reported in the literature. In this review we attempt to report on progress in the development of biologically derived systems able to show the phenomena of light amplification and random lasing together with the contribution of our group to this field. The rich world of biopolymers modified with molecular aggregates and nanocrystals, and self-organized at the nanoscale, offers a multitude of possibilities for tailoring luminescent and light scattering properties that are not easily replicated in conventional organic or inorganic materials. Of particular importance and interest are light amplification and lasing, or random lasing studies in biological cells and tissues. In this review we will describe nucleic acids and their complexes employed as gain media due to their favorable optical properties and ease of manipulation. We will report on research conducted on various biomaterials showing structural analogy to nucleic acids such as fluorescent proteins, gelatins in which the first distributed feedback laser was realized, and also amyloids or silks, which, due to their dye-doped fiber-like structure, allow for light amplification. Other materials that were investigated in that respect include polysaccharides, like starch exhibiting favorable photostability in comparison to other biomaterials, and chitosan, which forms photonic crystals or cellulose. Light amplification and random lasing was not only observed in processed biomaterials but also in living

  5. Improvement of Shelf Life and Sensory Quality of Pears Using a Specialized Edible Coating

    Directory of Open Access Journals (Sweden)

    Virgilio Cruz

    2015-01-01

    Full Text Available An edible coating functionalized with pomegranate polyphenols was designed. Different blends of candelilla wax, gum arabic, jojoba oil, and pomegranate polyphenols were formulated in order to improve the shelf life quality of pears (variety Bartlett, and all formulations were applied by immersion onto the fruit surface. Coated pears with and without polyphenols and uncoated pears (control were stored under the same conditions. Fruits were analyzed to evaluate changes in their physicochemical, microbiological, and sensorial properties during 30 days of storage at room temperature. Coated pears coded as T13 (candelilla wax 3%, gum arabic 4%, jojoba oil 0.15%, and pomegranate polyphenols 0.015% extended and improved their shelf life quality due to the minimization of the physic-chemical changes and sensorial properties. Therefore, the results indicated that the formulated edible coating has potential to extend the shelf life and maintain quality of pears. It was probed that coated pears were accepted for consumers as a good product. Edible coating application represents a good alternative to keep pears freshness for longer periods.

  6. Digital Microfluidics for Nucleic Acid Amplification

    Directory of Open Access Journals (Sweden)

    Beatriz Coelho

    2017-06-01

    Full Text Available Digital Microfluidics (DMF has emerged as a disruptive methodology for the control and manipulation of low volume droplets. In DMF, each droplet acts as a single reactor, which allows for extensive multiparallelization of biological and chemical reactions at a much smaller scale. DMF devices open entirely new and promising pathways for multiplex analysis and reaction occurring in a miniaturized format, thus allowing for healthcare decentralization from major laboratories to point-of-care with accurate, robust and inexpensive molecular diagnostics. Here, we shall focus on DMF platforms specifically designed for nucleic acid amplification, which is key for molecular diagnostics of several diseases and conditions, from pathogen identification to cancer mutations detection. Particular attention will be given to the device architecture, materials and nucleic acid amplification applications in validated settings.

  7. Stability of cactus-pear powder during storage

    Directory of Open Access Journals (Sweden)

    Plúvia O. Galdino

    2016-02-01

    Full Text Available ABSTRACT The stability of cactus-pear powder, obtained by the process of spray drying for 40 days, was evaluated under controlled conditions of relative air humidity (83% and temperature (25 and 40 °C. The whole pulp was characterized with regard to its physico-chemical parameters: pH, total titratable acidity, soluble solids, water content, total solids, ashes, reducing sugars, total sugars, non-reducing sugars, luminosity, redness, yellowness and water activity. The stored samples in powder were evaluated every 10 days for water content, water activity, total titratable acidity and color (luminosity, redness and yellowness. The whole pulp was slightly acidic and perishable, due to the high water content. During storage, the packages did not prevent water absorption, thus increasing water content and, consequently, water activity. Yellowness oscillated along the storage time, but the predominance of the yellow color was not affected.

  8. Studies of pear-shaped nuclei using accelerated radioactive beams

    CERN Document Server

    Gaffney, L P; Scheck, M; Hayes, A B; Wenander, F; Albers, M; Bastin, B; Bauer, C; Blazhev, A; Bonig, S; Bree, N; Cederkall, J; Chupp, T; Cline, D; Cocolios, T E; Davinson, T; DeWitte, H; Diriken, J; Grahn, T; Herzan, A; Huyse, M; Jenkins, D G; Joss, D T; Kesteloot, N; Konki, J; Kowalczyk, M; Kroll, Th; Kwan, E; Lutter, R; Moschner, K; Napiorkowski, P; Pakarinen, J; Pfeiffer, M; Radeck, D; Reiter, P; Reynders, K; Rigby, S V; Robledo, L M; Rudigier, M; Sambi, S; Seidlitz, M; Siebeck, B; Stora, T; Thoele, P; Van Duppen, P; Vermeulen, M J; von Schmid, M; Voulot, D; Warr, N; Wimmer, K; Wrzosek-Lipska, K; Wu, C Y; Zielinska, M

    2013-01-01

    There is strong circumstantial evidence that certain heavy, unstable atomic nuclei are ‘octupole deformed’, that is, distorted into a pear shape. This contrasts with the more prevalent rugby-ball shape of nuclei with reflection-symmetric, quadrupole deformations. The elusive octupole deformed nuclei are of importance for nuclear structure theory, and also in searches for physics beyond the standard model; any measurable electric-dipole moment (a signature of the latter) is expected to be amplified in such nuclei. Here we determine electric octupole transition strengths (a direct measure of octupole correlations) for short-lived isotopes of radon and radium. Coulomb excitation experiments were performed using accelerated beams of heavy, radioactive ions. Our data on and $^{224}$Ra show clear evidence for stronger octupole deformation in the latter. The results enable discrimination between differing theoretical approaches to octupole correlations, and help to constrain suitable candidates for experimental...

  9. Refillable and magnetically actuated drug delivery system using pear-shaped viscoelastic membrane

    KAUST Repository

    So, Hongyun; Seo, Young Ho; Pisano, Albert P.

    2014-01-01

    We report a refillable and valveless drug delivery device actuated by an external magnetic field for on-demand drug release to treat localized diseases. The device features a pear-shaped viscoelastic magnetic membrane inducing asymmetrical

  10. Empirical models in the description of prickly pear shoot (Nopal drying kinetics

    Directory of Open Access Journals (Sweden)

    Emmanuel M. Pereira

    Full Text Available ABSTRACT The objective of this study was to describe the technological process involved in the drying kinetics of fresh-cut prickly pear shoots through numerical and analytical solutions. Shoots of two different prickly pear species were used, ‘Gigante’ and ‘Miúda’. Drying was performed at different temperatures (50, 60, 70 and 80 °C and weighing procedures were made continuously. The experimental data were expressed as moisture ratio. The Page model showed the best fit to the drying kinetics of minimally processed ‘Gigante’ and ‘Miúda’ prickly pear shoots, with the best coefficients of determination and Chi-square. Peleg and Wang & Singh models can not be used to simulate the drying of ‘Gigante’ and ‘Miúda’ prickly pear shoots within the evaluated range of temperatures, showing an incoherent graphic pattern.

  11. Aromatically enhanced pear distillates from blanquilla and conference varieties using a packed column.

    Science.gov (United States)

    Arrieta-Garay, Yanine; García-Llobodanin, Laura; Pérez-Correa, José Ricardo; López-Vázquez, Cristina; Orriols, Ignacio; López, Francisco

    2013-05-22

    Pear distillates are generally produced from the Bartlett variety because of its rich aroma. In this study, a chemical and sensorial comparative examination of pear distillates from the three main varieties grown in Spain (Bartlett, Blanquilla, and Conference) using two distillation systems (copper Charentais alembic and packed column) was undertaken. Volatile compounds were identified by gas chromatography to differentiate the spirits according to pear variety and distillation method. The Bartlett distillates from both distillation systems possessed higher ethyl ester and acetate and lower cis-3-hexen-1-ol and 1-hexanol concentrations. Despite these differences, a sensory analysis panel could distinguish only the Bartlett alembic distillate from the alembic distillates of the other varieties. In contrast, the panel rated the packed-column distillates equally. Therefore, less aromatic pear varieties can be used to produce distillates with aromatic characteristics similar to those of the Bartlett variety if a suitable distillation process is used.

  12. 76 FR 4202 - Pears Grown in Oregon and Washington; Amendment To Allow Additional Exemptions

    Science.gov (United States)

    2011-01-25

    ... orchards, packing facilities, roadside stands, or farmers' markets without regard to the marketing order's... of pears sold at orchards, packing facilities, roadside stands, and farmers' markets should also be... directly to consumers from their [[Page 4203

  13. Mathematical modelling of thin layer drying of pear

    Directory of Open Access Journals (Sweden)

    Lutovska Monika

    2016-01-01

    Full Text Available In this study, a thin - layer drying of pear slices as a function of drying conditions were examined. The experimental data set of thin - layer drying kinetics at five drying air temperatures 30, 40, 50, 60 and 70°C, and three drying air velocities 1, 1.5 and 2 m s-1 were obtained on the experimental setup, designed to imitate industrial convective dryer. Five well known thin - layer drying models from scientific literature were used to approximate the experimental data in terms of moisture ratio. In order to find which model gives the best results, numerical experiments were made. For each model and data set, the statistical performance index, (φ, and chi-squared, (χ2, value were calculated and models were ranked afterwards. The performed statistical analysis shows that the model of Midilli gives the best statistical results. Because the effect of drying air temperature and drying air velocity on the empirical parameters was not included in the base Midilli model, in this study the generalized form of this model was developed. With this model, the drying kinetic data of pear slices can be approximated with high accuracy. The effective moisture diffusivity was determined by using Fick’s second laws. The obtained values of the effective moisture diffusivity, (Deff, during drying ranged between 6.49 x 10-9 and 3.29 x 10-8 m2 s-1, while the values of activation energy (E0 varied between 28.15 to 30.51 kJ mol-1.

  14. In-vitro neoformation of woody plants (apple, pear, Weigela)

    International Nuclear Information System (INIS)

    Chevreau, E.; Dufour, M.; Duron, M.

    1990-01-01

    Full text: Adventitious bud neoformation may induce variability in two ways: somaclonal variation, if the explant has undergone undifferentiated growth (callus); induced mutations, if a physical or chemical mutagen has been applied to the explant prior to the regeneration. Three apple scion cultivars ('Gala', 'Granny Smith', 'Golden Delicious') and four apple rootstock cultivars ('Mark', 'Novole', 'Lancep', 'Cepiland') were grown on basal apple multiplication medium. Leaves, internodes and roots were used for callogenesis and/or regeneration experiments. Scion varieties were more responsive than rootstocks. 1000 'Gala' neoformations will be planted in the field to investigate somaclonal variation. In pear, shoot regeneration was obtained from in-vitro leaves of 3 varieties of Pyrus communis ('Seckel', 'Louise Bonne', 'Comice') and one variety of Pyrus bretschneideri ('Crystal Pear'). It was observed that regeneration is still possible from leaves irradiated with up to 40 Gy of γ rays, or treated with 5mM ENU for one hour. In Weigela, stem segments (0.5 cm) from rooted in vitro plants of 5 cultivars were used in experiments to induce adventitious buds. On the cultivar 'Eva Rathke' the adventitious buds were detected as early as 10 to 15 days after the beginning of the culture. A mutagenic treatment was carried out by immersing explants of 'Bristol Ruby' in a suspension of ethyl methane sulfonate (EMS). Among 400 plants regenerated from treated explants, 5 mutants were detected after 5 years of field observation. Two of them seem to be solid mutants. Apparently in Weigela both homogeneous mutants and chimeras are produced by adventitious regeneration from mutagenised explants. (author)

  15. Occurrence of Anthracnose on Fruits of Asian Pear Tree Caused by Colletotrichum acutatum

    OpenAIRE

    Kim, Wan Gyu; Hong, Sung Kee; Park, Yeong Seob

    2007-01-01

    Anthracnose symptoms often occurred on fruits of Asian pear trees grown in Anseong, Naju, Seonghwan and Pyeongtaek in Korea during the harvesting period from 2000 to 2005. A total of 28 isolates of Colletotrichum sp. were obtained from the anthracnose symptoms. All the isolates were identified as Colletotrichum acutatum based on their morphological and cultural characteristics. Four isolates of the fungus were tested for pathogenicity to fruits of Asian pear tree by artificial inoculation. Al...

  16. Cell-Specific PEAR1 Methylation Studies Reveal a Locus that Coordinates Expression of Multiple Genes

    Directory of Open Access Journals (Sweden)

    Benedetta Izzi

    2018-04-01

    Full Text Available Chromosomal interactions connect distant enhancers and promoters on the same chromosome, activating or repressing gene expression. PEAR1 encodes the Platelet-Endothelial Aggregation Receptor 1, a contact receptor involved in platelet function and megakaryocyte and endothelial cell proliferation. PEAR1 expression during megakaryocyte differentiation is controlled by DNA methylation at its first CpG island. We identified a PEAR1 cell-specific methylation sensitive region in endothelial cells and megakaryocytes that showed strong chromosomal interactions with ISGL20L2, RRNAD1, MRLP24, HDGF and PRCC, using available promoter capture Hi-C datasets. These genes are involved in ribosome processing, protein synthesis, cell cycle and cell proliferation. We next studied the methylation and expression profile of these five genes in Human Umbilical Vein Endothelial Cells (HUVECs and megakaryocyte precursors. While cell-specific PEAR1 methylation corresponded to variability in expression for four out of five genes, no methylation change was observed in their promoter regions across cell types. Our data suggest that PEAR1 cell-type specific methylation changes may control long distance interactions with other genes. Further studies are needed to show whether such interaction data might be relevant for the genome-wide association data that showed a role for non-coding PEAR1 variants in the same region and platelet function, platelet count and cardiovascular risk.

  17. PEAR: a fast and accurate Illumina Paired-End reAd mergeR.

    Science.gov (United States)

    Zhang, Jiajie; Kobert, Kassian; Flouri, Tomáš; Stamatakis, Alexandros

    2014-03-01

    The Illumina paired-end sequencing technology can generate reads from both ends of target DNA fragments, which can subsequently be merged to increase the overall read length. There already exist tools for merging these paired-end reads when the target fragments are equally long. However, when fragment lengths vary and, in particular, when either the fragment size is shorter than a single-end read, or longer than twice the size of a single-end read, most state-of-the-art mergers fail to generate reliable results. Therefore, a robust tool is needed to merge paired-end reads that exhibit varying overlap lengths because of varying target fragment lengths. We present the PEAR software for merging raw Illumina paired-end reads from target fragments of varying length. The program evaluates all possible paired-end read overlaps and does not require the target fragment size as input. It also implements a statistical test for minimizing false-positive results. Tests on simulated and empirical data show that PEAR consistently generates highly accurate merged paired-end reads. A highly optimized implementation allows for merging millions of paired-end reads within a few minutes on a standard desktop computer. On multi-core architectures, the parallel version of PEAR shows linear speedups compared with the sequential version of PEAR. PEAR is implemented in C and uses POSIX threads. It is freely available at http://www.exelixis-lab.org/web/software/pear.

  18. Revalorization of cactus pear (Opuntia spp. wastes as a source of antioxidants

    Directory of Open Access Journals (Sweden)

    Anaberta Cardador-Martínez

    2011-09-01

    Full Text Available Recently, an increased interest in antioxidant activity and health-improving capacity of cactus pear has been registered. The antioxidant capacity of the pulp of cactus-pear fruits has been previously assessed. In this work, total phenolics, flavonoids and tannins of peel and seeds of four cactus pear cultivars were examined as well as their antioxidant capacity. Tannins were the major phenolics in cactus pear seeds accounting for almost fifty percent for all cultivars. Analysis of variance revealed that ripeness, cultivar, and its interaction had highly significant effect on the total phenolics, tannin, and flavonoid contents of cactus pear peel. With regard to the seeds, only the stage of ripeness and interaction (ripeness stage x cultivar were significant on total phenolics and tannins contents. The flavonoid content in seeds was not affected by any of the factors or their interactions. The antioxidant capacity was higher in the peel than in the seeds. Generally, fruits with light-green or yellow-brown peel have higher antiradical activity and Trolox equivalent antioxidant capacity (TEAC values compared with those with red-purple peel. Cactus pear by-products can indeed be exploited as a good and cheap source of natural antioxidants.

  19. Population diversity of ammonium oxidizers investigated by specific PCR amplification

    Science.gov (United States)

    Ward, B.B.; Voytek, M.A.; Witzel, K.-P.

    1997-01-01

    The species composition of ammonia-oxidizing bacteria in aquatic environments was investigated using PCR primers for 16S rRNA genes to amplify specific subsets of the total ammonia-oxidizer population. The specificity of the amplification reactions was determined using total genomic DNA from known nitrifying strains and non-nitrifying strains identified as having similar rDNA sequences. Specificity of amplification was determined both for direct amplification, using the nitrifier specific primers, and with nested amplification, in which the nitrifier primers were used to reamplify a fragment obtained from direct amplification with Eubacterial universal primers. The present level of specificity allows the distinction between Nitrosomonas europaea, Nitrosomonas sp. (marine) and the other known ammonia-oxidizers in the beta subclass of the Proteobacteria. Using total DNA extracted from natural samples, we used direct amplification to determine presence/absence of different species groups. Species composition was found to differ among depths in vertical profiles of lake samples and among samples and enrichments from various other aquatic environments. Nested PCR yielded several more positive reactions, which implies that nitrifier DNA was present in most samples, but often at very low levels.

  20. Evidence of high-elevation amplification versus Arctic amplification.

    Science.gov (United States)

    Wang, Qixiang; Fan, Xiaohui; Wang, Mengben

    2016-01-12

    Elevation-dependent warming in high-elevation regions and Arctic amplification are of tremendous interest to many scientists who are engaged in studies in climate change. Here, using annual mean temperatures from 2781 global stations for the 1961-2010 period, we find that the warming for the world's high-elevation stations (>500 m above sea level) is clearly stronger than their low-elevation counterparts; and the high-elevation amplification consists of not only an altitudinal amplification but also a latitudinal amplification. The warming for the high-elevation stations is linearly proportional to the temperature lapse rates along altitudinal and latitudinal gradients, as a result of the functional shape of Stefan-Boltzmann law in both vertical and latitudinal directions. In contrast, neither altitudinal amplification nor latitudinal amplification is found within the Arctic region despite its greater warming than lower latitudes. Further analysis shows that the Arctic amplification is an integrated part of the latitudinal amplification trend for the low-elevation stations (≤500 m above sea level) across the entire low- to high-latitude Northern Hemisphere, also a result of the mathematical shape of Stefan-Boltzmann law but only in latitudinal direction.

  1. Back-transmission of a virus associated with apple stem pitting and pear vein yellows from Nicotiana occidentalis to apple and pear indicators

    NARCIS (Netherlands)

    Leone, G.; Lindner, J.L.; Jongedijk, G.; Meer, van der F.

    1995-01-01

    The successful back-transmission of the mechanically transmissible virus associated with apple stem pitting and pear vein yellows, from Nicotiana occidentalis to apple seedlings "Golden Delicious" under greenhouse conditions is reported. This result enabled a field experiment where isolates of apple

  2. Symptoms on apple and pear indicators after back-transmission from Nicotiana occidentalis confirm the identity of apple stem pitting virus with pear vein yellows virus

    NARCIS (Netherlands)

    Leone, G.; Lindner, J.L.; Meer, van der F.A.; Schoen, C.D.; Jongedijk, G.

    1998-01-01

    Isolates of apple stem pitting virus (ASPV) from diseased apple trees were maintained in Nicotiana occidentalis then back-transmitted mechanically from the herbaceous host to apple seedlings and indexed by double budding on apple and pear indicators for the following syndromes: apple stem pitting,

  3. Efficient Audio Power Amplification - Challenges

    DEFF Research Database (Denmark)

    Andersen, Michael Andreas E.

    2005-01-01

    For more than a decade efficient audio power amplification has evolved and today switch-mode audio power amplification in various forms are the state-of-the-art. The technical steps that lead to this evolution are described and in addition many of the challenges still to be faced and where...

  4. Efficient audio power amplification - challenges

    Energy Technology Data Exchange (ETDEWEB)

    Andersen, Michael A.E.

    2005-07-01

    For more than a decade efficient audio power amplification has evolved and today switch-mode audio power amplification in various forms are the state-of-the-art. The technical steps that lead to this evolution are described and in addition many of the challenges still to be faced and where extensive research and development are needed is covered. (au)

  5. Proteomic analysis of 'Zaosu' pear (Pyrus bretschneideri Rehd.) and its early-maturing bud sport.

    Science.gov (United States)

    Liu, Xueting; Zhai, Rui; Feng, Wenting; Zhang, Shiwei; Wang, Zhigang; Qiu, Zonghao; Zhang, Junke; Ma, Fengwang; Xu, Lingfei

    2014-07-01

    Maturation of fruits involves a series of physiological, biochemical, and organoleptic changes that eventually make fleshy fruits attractive, palatable, and nutritional. In order to understand the mature mechanism of the early-maturing bud sport of 'Zaosu' pear, we analyzed the differences of proteome expression between the both pears in different mature stages by the methods of a combination of two-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis. Seventy-five differential expressed protein spots (psport, but only sixty-eight were demonstratively identified in the database of NCBI and uniprot. The majority of proteins were linked to metabolism, energy, stress response/defense and cell structure. Additionally, our data confirmed an increase of proteins related to cell-wall modification, oxidative stress and pentose phosphate metabolism and a decrease of proteins related to photosynthesis and glycolysis during the development process of both pears, but all these proteins increased or decreased faster in the early-maturing bud sport. This comparative analysis between both pears showed that these proteins were closely associated with maturation and could provide more detailed characteristics of the maturation process of both pears. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  6. Effects of ultrasound treatment in purple cactus pear (Opuntia ficus-indica) juice.

    Science.gov (United States)

    Zafra-Rojas, Quinatzin Yadira; Cruz-Cansino, Nelly; Ramírez-Moreno, Esther; Delgado-Olivares, Luis; Villanueva-Sánchez, Javier; Alanís-García, Ernesto

    2013-09-01

    Cactus pear (Opuntia ficus-indica) fruit is a berry with a tasty pulp full of seeds that constitutes about 10-15% of the edible pulp. In Mexico, cactus pear is mainly consumed fresh, but also has the potential to be processed in other products such as juice. The objective of this study was to evaluate the effect of different ultrasound conditions at amplitude levels ranging (40% and 60% for 10, 15, 25 min; 80% for 3, 5, 8, 10, 15 and 25 min) on the characteristics of purple cactus pear juice. The evaluated parameters were related with the quality (stability, °Brix, pH), microbial growth, total phenolic compounds, ascorbic acid and antioxidant activity (ABTS, DPPH and % chelating activity) of purple cactus pear juices. The ultrasound treatment for time period of 15 and 25 min significantly reduced the microbial count in 15 and 25 min, without affecting the juice quality and its antioxidant properties. Juice treated at 80% of amplitude level showed an increased of antioxidant compounds. Our results demonstrated that sonication is a suitable technique for cactus pear processing. This technology allows the achievement of juice safety and quality standards without compromising the retention of antioxidant compounds. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Apomixis and the problem of obtaining haploids and homozygote diploids in pear (Pyrus communis L.

    Directory of Open Access Journals (Sweden)

    Є. О. Долматов

    2013-02-01

    Full Text Available The article highlights results of research over simulative apomixes in pear and its utilization for obtaining haploids and homozygote diploids. It has been established that over 50% pear varieties with failed remote hybridization are capable of generating seeds of apomictic origin producing diploid plants. Genotypes displaying maximal inclination to apomixes have been singled out. Apomictic pear seedlings obtained from foreign pollination within the limits of the same combination are inherent in profound morphological diversity. Fruit-bearing apomicts originated from one and the same maternal plant differ to the same extent as hybrid seedlings of the same family. Genetic markers have enabled to establish that these are embryo sacs in which meiosis has completed that give rise to apomictic seeds. In vitro method as used for the purpose of increasing apomictic plants output has been illustrated. The greatest induction of apomictic shoots in vitro has been reached by alternation of BAP cytokinin at concentration of 1mg/l and 2 mg/l on the background of GA3 amounting to 1,5 mg/l. Grafting with shoots in vitro on non-sterile rootstocks of pear (Pyrus communis has increased the output of plants up to 80%. A cytological assessment of 9 apomictic samples is provided. The cytological analysis of samples of apomictic forms has certified the presence of simulative haploid parthenogenesis in pear.

  8. RNA amplification for successful gene profiling analysis

    Directory of Open Access Journals (Sweden)

    Wang Ena

    2005-07-01

    Full Text Available Abstract The study of clinical samples is often limited by the amount of material available to study. While proteins cannot be multiplied in their natural form, DNA and RNA can be amplified from small specimens and used for high-throughput analyses. Therefore, genetic studies offer the best opportunity to screen for novel insights of human pathology when little material is available. Precise estimates of DNA copy numbers in a given specimen are necessary. However, most studies investigate static variables such as the genetic background of patients or mutations within pathological specimens without a need to assess proportionality of expression among different genes throughout the genome. Comparative genomic hybridization of DNA samples represents a crude exception to this rule since genomic amplification or deletion is compared among different specimens directly. For gene expression analysis, however, it is critical to accurately estimate the proportional expression of distinct RNA transcripts since such proportions directly govern cell function by modulating protein expression. Furthermore, comparative estimates of relative RNA expression at different time points portray the response of cells to environmental stimuli, indirectly informing about broader biological events affecting a particular tissue in physiological or pathological conditions. This cognitive reaction of cells is similar to the detection of electroencephalographic patterns which inform about the status of the brain in response to external stimuli. As our need to understand human pathophysiology at the global level increases, the development and refinement of technologies for high fidelity messenger RNA amplification have become the focus of increasing interest during the past decade. The need to increase the abundance of RNA has been met not only for gene specific amplification, but, most importantly for global transcriptome wide, unbiased amplification. Now gene

  9. First Report of Cadophora luteo-olivacea Causing Side Rot on ‘Conference’ Pears in the Netherlands

    NARCIS (Netherlands)

    Wenneker, M.; Pham, K.T.K.; Lemmers, M.E.C.; Boer, de Astrid; Leeuwen, van Paul; Hollinger, T.C.; Haas, de B.H.; Köhl, J.

    2016-01-01

    Pear (Pyrus communis) is an important fruit crop in the Netherlands. Symptoms of side rot disease of pear fruits were first observed in 2008 on cv. Conference in storage in the Netherlands. Typical round to oval, dark-brown, and slightly sunken spots (size 0.5 to 1.0 cm in diameter) appeared after

  10. AGROCLIMATIC ZONING OF EUROPEAN AND ASIAN PEAR CULTIVARS WITH POTENTIAL FOR COMMERCIAL PLANTING IN SOUTHERN BRAZIL

    Directory of Open Access Journals (Sweden)

    MARCOS SILVEIRA WREGE

    Full Text Available ABSTRACT Pear is among the fruits of major commercial interest in the world and one of the most imported in Brazil. Brazilian production is very small and the fruit quality is low, due to production problems. The success of culture in the country, among other factors, may be linked to the choice of cultivars, pollinating and rootstocks better adapted to local conditions, and thus depend on the particulars of an agricultural zoning. The aim of this study was to identify, in southern Brazil, homogeneous climatic zones with potential for growing of European and Asian pears, through climate risk studies. The regions were defined by the seasonal availability of chilling hours (<7.2 °C accumulated during the period from May to September and the monthly risk of drought. The analysis allowed the recognition of four homogeneous areas for the production of pears in southern Brazil.

  11. Effect of Root Pruning and Irrigation Regimes on Yield and Physiology of Pear Trees

    DEFF Research Database (Denmark)

    Wang, Yufei

    Clara Frijs’ is the dominant pear (Pyrus communis L.) cultivar in Denmark. It is vigorous with long annual shoots, and therefore can be difficult to prune. Root pruning has been widely used to control the canopy size of fruit trees including pears. However, root pruned trees are more likely......, it was concluded that root pruning not only decreases water uptake but also nutrient uptake, and both have contributed to the reduced canopy growth. Supplemental irrigation partially improved the tree water status and nitrogen uptake without stimulating additional shoot growth in the root pruned trees....... A combination of root pruning and irrigation could be a promising practice to control tree size and secure a stable fruit yield in pear orchard....

  12. Potable NIR spectroscopy predicting soluble solids content of pears based on LEDs

    Energy Technology Data Exchange (ETDEWEB)

    Liu Yande; Liu Wei; Sun Xudong; Gao Rongjie; Pan Yuanyuan; Ouyang Aiguo, E-mail: jxliuyd@163.com [School of Mechatronics Engineering, East China Jiaotong University, Changbei Open and Developing District, Nanchang, 330013 (China)

    2011-01-01

    A portable near-infrared (NIR) instrument was developed for predicting soluble solids content (SSC) of pears equipped with light emitting diodes (LEDs). NIR spectra were collected on the calibration and prediction sets (145:45). Relationships between spectra and SSC were developed by multivariate linear regression (MLR), partial least squares (PLS) and artificial neural networks (ANNs) in the calibration set. The 45 unknown pears were applied to evaluate the performance of them in terms of root mean square errors of prediction (RMSEP) and correlation coefficients (r). The best result was obtained by PLS with RMSEP of 0.62{sup 0}Brix and r of 0.82. The results showed that the SSC of pears could be predicted by the portable NIR instrument.

  13. Potable NIR spectroscopy predicting soluble solids content of pears based on LEDs

    International Nuclear Information System (INIS)

    Liu Yande; Liu Wei; Sun Xudong; Gao Rongjie; Pan Yuanyuan; Ouyang Aiguo

    2011-01-01

    A portable near-infrared (NIR) instrument was developed for predicting soluble solids content (SSC) of pears equipped with light emitting diodes (LEDs). NIR spectra were collected on the calibration and prediction sets (145:45). Relationships between spectra and SSC were developed by multivariate linear regression (MLR), partial least squares (PLS) and artificial neural networks (ANNs) in the calibration set. The 45 unknown pears were applied to evaluate the performance of them in terms of root mean square errors of prediction (RMSEP) and correlation coefficients (r). The best result was obtained by PLS with RMSEP of 0.62 0 Brix and r of 0.82. The results showed that the SSC of pears could be predicted by the portable NIR instrument.

  14. Occurrence of Anthracnose on Fruits of Asian Pear Tree Caused by Colletotrichum acutatum.

    Science.gov (United States)

    Kim, Wan Gyu; Hong, Sung Kee; Park, Yeong Seob

    2007-12-01

    Anthracnose symptoms often occurred on fruits of Asian pear trees grown in Anseong, Naju, Seonghwan and Pyeongtaek in Korea during the harvesting period from 2000 to 2005. A total of 28 isolates of Colletotrichum sp. were obtained from the anthracnose symptoms. All the isolates were identified as Colletotrichum acutatum based on their morphological and cultural characteristics. Four isolates of the fungus were tested for pathogenicity to fruits of Asian pear tree by artificial inoculation. All the isolates induced anthracnose symptoms on the fruits by wound inoculation but not by unwound inoculation. The anthracnose symptoms induced by artificial inoculation were similar to those observed in the orchard. This is the first report of anthracnose of Asian pear tree caused by Colletotrichum acutatum.

  15. Conservation of Williams pear using edible coating with alginate and carrageenan

    Directory of Open Access Journals (Sweden)

    Kessiane Silva de Moraes

    2012-12-01

    Full Text Available The aim of this study was to evaluate the physical and chemical parameters of Williams pear, stored at 25 ºC for 15 days, with and without edible coating. Edible coatings prepared with alginate 2% and carrageenan 0.5% were tested. The analyses carried out on the samples were: weight loss, pH, soluble solids, firmness, and color. The edible coatings were characterized in terms of mechanical properties, permeability, thickness, and opacity. The results show that the application of edible coatings with carrageenan and alginate in pears influenced physical and chemical characteristics such as weight loss, pH, total soluble solids, color, and firmness of the fruit. However, the alginate coating showed the best results on pear conservation since it had lower water vapor permeability and greater tensile strength, and therefore it can be used as a protective film on these fruits.

  16. Utilization of Prickly Pear Peels to Improve Quality of Pan Bread

    International Nuclear Information System (INIS)

    Anwar, M.M.; Sallam, E.M.

    2016-01-01

    This investigation aimed to study utilization of prickly pear peels to improve quality of pan bread. Prickly pear peels powder added to wheat flour 72 % at levels 1.0 and 2.0% to make pan beard. In this study, evaluation of nutrients and chemical constitutes and functional properties of prickly pear peels as well as the rheological properties of dough contained prickly pear peels at levels 1% and 2% has been conducted. Then evaluated organoleptic characteristics of pan bread made of it also determined staling rate. Results showed that prickly pear peels had higher content of 32.67 % fiber, 14.25 % pectin and 87.82 % ascorbic acid, and higher contents of antioxidant components. It consists of 441.11 mg/100 g, total phenols, 35.2 flavonoids mg/100 g and DPPH radical-scavenging 62.14%, also water holding capacity was 1.8 ml H 2 O / g , oil holding capacity 2.35 (ml oil/g) and foam stability 7.15%. The major phenolic compounds Oleuro 1264.407, pyrogallo 1149.68, Benzoic 982.37, 3-oH Tyrosol 588.53, Ellagic 413.26, Chorogenic 271.10, Protocatechuic acid 176.02, P-oH- Benzoic 112.78, Epicatechin105.99, Gallic acid 61.26 ppm. The results revealed that addition of prickly pear peels to wheat flour increased the nutrition values of pan bread made of it due to high contents of fiber, ascorbic acid and natural antioxidants, and also decreased staling which improves the quality of pan bread, as well as increases shelf-life of pan bread.

  17. Controlled Microwave Heating Accelerates Rolling Circle Amplification.

    Directory of Open Access Journals (Sweden)

    Takeo Yoshimura

    Full Text Available Rolling circle amplification (RCA generates single-stranded DNAs or RNA, and the diverse applications of this isothermal technique range from the sensitive detection of nucleic acids to analysis of single nucleotide polymorphisms. Microwave chemistry is widely applied to increase reaction rate as well as product yield and purity. The objectives of the present research were to apply microwave heating to RCA and indicate factors that contribute to the microwave selective heating effect. The microwave reaction temperature was strictly controlled using a microwave applicator optimized for enzymatic-scale reactions. Here, we showed that microwave-assisted RCA reactions catalyzed by either of the four thermostable DNA polymerases were accelerated over 4-folds compared with conventional RCA. Furthermore, the temperatures of the individual buffer components were specifically influenced by microwave heating. We concluded that microwave heating accelerated isothermal RCA of DNA because of the differential heating mechanisms of microwaves on the temperatures of reaction components, although the overall reaction temperatures were the same.

  18. Changes in the physiological characteristics of mature pear fruits subjected to gamma irradiation

    International Nuclear Information System (INIS)

    Mahmoud, A.A.; Sewidan, A.M.; Roushdy, H.M.; Taha, F.

    1988-01-01

    Moisture content, weight loss, firmness, titrable acidity total soluble solids and decay percentage of the pear fruits (variety of Le conte) of different treatments (0,1,2,3, and 4 KGy) were investigated during storage. The data reported show that there was a slight decrease in the moisture content and titrable acidity of pear fruits during storage and there was decrease in the firmness and decay percentage. This decrease was elevated by the increase of irradiation doses and storage period on the other hand, the weight losses and total soluble solids increased by increasing the irradiation doses and storage period

  19. Amplification of the dd reaction and X radiation generation in a high-current pulse glow discharge in deuterium with the Ti cathode at 0.8-2.45 kV

    International Nuclear Information System (INIS)

    Lipson, A.G.; Rusetskij, A.S.; Karabut, A.B.; Majli, Dzh.

    2005-01-01

    The dd-reaction yield (3 MeV protons) and the soft X-ray emission from a titanium cathode surface in a periodic pulsed glow discharge in deuterium were studied. The deuteron screening potential was estimated from analysis of the dd-reaction yield as a function of the accelerating voltage. The obtained data show evidence for a significant enhancement of the dd-reaction yield in Ti in comparison to both theoretical estimates and the results of experiments using accelerators at the deuteron energies ≥ 2.5 keV. Intense emission of soft X-ray quanta was observed. The X-ray emission intensity and the dd-reaction yield enhancement strongly depend on the rate of deuterium diffusion in a thin subsurface layer of Ti cathode [ru

  20. Next generation Chirped Pulse Amplification

    Energy Technology Data Exchange (ETDEWEB)

    Nees, J; Biswal, S; Mourou, G [Univ. Michigan, Center for Ultrafast Optical Science, Ann Arbor, MI (United States); Nishimura, Akihiko; Takuma, Hiroshi

    1998-03-01

    The limiting factors of Chirped Pulse Amplification (CPA) are discussed and experimental results of CPA in Yb:glass regenerative amplifier are given. Scaling of Yb:glass to the petawatt level is briefly discussed. (author)

  1. Amplification of Chirality through Self-Replication of Micellar Aggregates in Water

    KAUST Repository

    Bukhriakov, Konstantin; Almahdali, Sarah; Rodionov, Valentin

    2015-01-01

    We describe a system in which the self-replication of micellar aggregates results in a spontaneous amplification of chirality in the reaction products. In this system, amphiphiles are synthesized from two "clickable" fragments: a water-soluble "head

  2. Rapid and Sensitive Isothermal Detection of Nucleic-acid Sequence by Multiple Cross Displacement Amplification

    OpenAIRE

    Yi Wang; Yan Wang; Ai-Jing Ma; Dong-Xun Li; Li-Juan Luo; Dong-Xin Liu; Dong Jin; Kai Liu; Chang-Yun Ye

    2015-01-01

    We have devised a novel amplification strategy based on isothermal strand-displacement polymerization reaction, which was termed multiple cross displacement amplification (MCDA). The approach employed a set of ten specially designed primers spanning ten distinct regions of target sequence and was preceded at a constant temperature (61?65??C). At the assay temperature, the double-stranded DNAs were at dynamic reaction environment of primer-template hybrid, thus the high concentration of primer...

  3. Complete Genome Sequence of Bacillus velezensis L-1, Which Has Antagonistic Activity against Pear Diseases

    OpenAIRE

    Sun, Pingping; Cui, Jianchao; Jia, Xiaohui; Wang, Wenhui

    2017-01-01

    ABSTRACT Bacillus velezensis L-1 is an effective biocontrol agent against pear diseases. Here, we report the complete genome sequence of B. velezensis L-1 in which clusters related to the biosynthesis of secondary metabolites were predicted. This genome provides insights into the possible biocontrol mechanisms and furthers application of this specific bacterium.

  4. Complete Genome Sequence of Bacillus velezensis L-1, Which Has Antagonistic Activity against Pear Diseases.

    Science.gov (United States)

    Sun, Pingping; Cui, Jianchao; Jia, Xiaohui; Wang, Wenhui

    2017-11-30

    Bacillus velezensis L-1 is an effective biocontrol agent against pear diseases. Here, we report the complete genome sequence of B. velezensis L-1 in which clusters related to the biosynthesis of secondary metabolites were predicted. This genome provides insights into the possible biocontrol mechanisms and furthers application of this specific bacterium. Copyright © 2017 Sun et al.

  5. Volatiles from Psylla-infested pear trees and their possible involvement in attraction of anthocorid predators

    NARCIS (Netherlands)

    Scutareanu, P.; Drukker, B.; Bruin, J.; Posthumus, M.A.; Sabelis, M.W.

    1997-01-01

    Previous work showed that anthocorid predators aggregate around gauze cages containing Psylla-infested trees in a pear orchard. Because anthocorids responded to odor from Psylla-infested leaves in a laboratory test, it was hypothesized that these aggregative responses in the field were triggered by

  6. Evaluation of Dacryodes edulis (native pear) seed biomass for Pb (II ...

    African Journals Online (AJOL)

    Dacryodes edulis (Native pear) seed is herein evaluated as a promising biomass for Pb(II) removal from aqueous solution. The Pb(II) adsorption onto Dacryodes edulis seed biomass was influenced by the solution pH, time, biomass dose and initial adsorbate concentration. The Freundlich model fitted better than the ...

  7. 7 CFR 319.56-29 - Ya variety pears from China.

    Science.gov (United States)

    2010-01-01

    ... conditions. (1) The pears must have been grown by growers registered with the national plant protection...) Field inspections for signs of pest infestation must be conducted by the national plant protection...-29 Agriculture Regulations of the Department of Agriculture (Continued) ANIMAL AND PLANT HEALTH...

  8. Development of an energy-protein for animal food based crop residues pear (Pyrus communis

    Directory of Open Access Journals (Sweden)

    Néstor Julián Pulido-Suárez,

    2016-01-01

    Full Text Available Pear (Pyrus communis is a fruit from the species of deciduous, widely consumed worldwide for its high quality energ y. However, pear itself does not provide the amount of protein required for cattle feeding, so alternatives to improve its nutritional quality have been studied. On these grounds, the objective of this study was to evaluate the parameters of solid state fermentation, and compositional energ y value of a protein food based on pears (Pyrus communis with apparent physical damage. A completely random design was used to evaluate three treatments; these correspond to percentages of inclusion of calcium carbonate (0.25, 0.50, 0.75 formulation based on already established (40 % pear, 25 % rice flour, 25 % wheat bran and 10 % urea, the parameters evaluated were: pH, ashes (CZ, crude protein (CP and crude fiber (CF, and they were recorded at 0, 24, 48 and 72 hours. As a result, it was found that the pH dropped gradually for each treatment and at each sampling period; however, there were no significant differences. The lower value at the end of the process is recorded T2 (0.25 with 4.66, followed by T3 (0.50 with 4.50, the ash reached values of up to 6 % with T3, and T2 (0.50 reached the highest percentages in fiber and crude protein. Finally, decreasing the fermentation variables ensures a food with no presence of undesirable microorganisms and stable over time.

  9. Relative Susceptibility of Quince, Pear, and Apple Cultivars to Fire Blight Following Greenhouse Inoculation

    Science.gov (United States)

    Fire blight caused by Erwinia amylovora (EA) is one of the most serious diseases of plants in the family Rosaceae, and Quince (Cydonia oblonga Mill.) is considered one of the most susceptible host genera. Apple (Malus sp.) and pear (Pyrus sp.) cultivars ranging from most susceptible to most resistan...

  10. Plant growth responses of apple and pear trees to doses of glyphosate

    Science.gov (United States)

    Glyphosate is commonly used for intra-row weed management in perennial plantations, where unintended crop exposure to this herbicide can cause growth reduction. The objective of this research was to analyze the initial plant growth behavior of young apple and pear plants exposed to glyphosate. Glyph...

  11. The genome of the pear (Pyrus bretschneideri Rehd.)

    DEFF Research Database (Denmark)

    Wu, Jun; Wang, Zhiwen; Shi, Zebin

    2013-01-01

    The draft genome of the pear (Pyrus bretschneideri) using a combination of BAC-by-BAC and next-generation sequencing is reported. A 512.0-Mb sequence corresponding to 97.1% of the estimated genome size of this highly heterozygous species is assembled with 194× coverage. High-density genetic maps ...

  12. EARLY SPRING APPLICATION OF AMINOETHOXYVINILGLYCINE (AVG INCREASES FRUIT SET AND YIELD OF ‘ROCHA’ PEARS

    Directory of Open Access Journals (Sweden)

    MATEUS DA SILVEIRA PASA

    2017-10-01

    Full Text Available ABSTRACT The low fruit set is one of the main factors leading to poor yield of pear orchards in Brazil. Ethylene is associated with abscission of flowers and fruitlets. Then, the application of ethylene synthesis inhibitors, such as AVG, is a potential tool to increase fruit set of pears. The objective of this study was to evaluate the effect of AVG, sprayed at different rates and timings, on fruit set, yield and fruit quality of ‘Rocha’ pear. The study was performed in a commercial orchard located in the municipality of São Joaquim, SC, during the growing seasons of 2014/2015 and 2015/2016. Plant material consisted of ‘Rocha’ pear trees grafted on quince rootstock ‘BA29’. AVG was tested at different rates (60 mg L-1 and 80 mg L-1 and timings [full bloom, one week after full bloom (WAFB, and two WAFB, either alone or in combination. The experiment was arranged in a randomized block design, with at least five single-tree replications. The fruit set, number of fruit per tree, yield, estimated yield, fruit weight, return bloom, and fruit quality attributes were assessed. Fruit set and yield were consistently increased by single applications of AVG at 60 and 80 mg L-1 at both one and two weeks after full bloom, without negatively affecting fruit quality attributes and return bloom.

  13. Estimate of Leaf Chlorophyll and Nitrogen Content in Asian Pear (Pyrus serotina Rehd. by CCM-200

    Directory of Open Access Journals (Sweden)

    Mostafa GHASEMI

    2011-03-01

    Full Text Available In many cases evaluation of chlorophyll and nitrogen content in plants need to destructive methods, more time and organic solvents. Application of chlorophyll meters save time and resources. The aim of this study was estimating of chlorophyll and nitrogen content in Asian pear leaves using non-destructive method and rapid quantification of chlorophyll by chlorophyll content meter (CCM-200. This study was conducted on 8 years old Asian pear trees during June 2008 in Tehran, Iran. To develop our regression model, the chlorophyll meter data were correlated with extracted chlorophyll and nitrogen content data obtained from DMSO and Kejeldal methods, respectively. The results showed that, there was positive and linear correlation between CCM-200 data and chlorophyll a (R�=0.7183, chlorophyll b (R�=0.8523, total chlorophyll (R�=0.90, and total nitrogen content (R�=0.76 in Asian pear leaves. Thus, it can be concluded that, CCM-200 can be used in order to predict both chlorophyll and nitrogen content in Asian pear leaves.

  14. Finite element analysis of the dynamic behavior of pear under impact loading

    Directory of Open Access Journals (Sweden)

    Alireza Salarikia

    2017-03-01

    Full Text Available Pear fruit is susceptible to bruising from mechanical impact during field harvesting operations and at all stages of postharvest handling. The postharvest shelf life of bruised fruits were shorter, and they softened rapidly under cold storage compared with non-bruised samples. Developing strategies for reducing bruising during the supply chain requires an understanding of fruit dynamic behavior to different enforced loadings. Finite Element Method (FEM is among the best techniques, in terms of accuracy and cost-efficiency, for studying the factors effective in impact-induced bruising. In this research, the drop test of pear sample was simulated using FEM. The simulation was conducted on a 3D solid model of the pear that was created by using non-contact optical scanning technology. This computer-based study aimed to assess the stress and strain distribution patterns within pear generated by collision of the fruit with a flat surface made of different materials. The contact force between two colliding surfaces is also investigated. The simulations were conducted at two different drop orientations and four different impact surfaces. Results showed that, in both drop orientations, the largest and smallest stresses, strains and contact forces were developed in collision with the steel and rubber surfaces, respectively. In general, these parameters were smaller when fruit collided with the surfaces along its horizontal axis than when collided along its vertical axis. Finally, analyses of stress and strain magnitudes showed that simulation stress and strain values were compatible with experiments data.

  15. Xylella taiwanensis sp. nov. cause of pear leaf scorch disease in Taiwan

    Science.gov (United States)

    Xylella fastidiosa is a group of xylem-limited and nutritionally fastidious plant pathogenic bacteria. While mostly found in the Americas, new X. fastidiosa strains have been reported from other continents such as Asia, including a pear leaf scorch (PLS) strain from Taiwan. Current taxonomy of X. fa...

  16. Soil sampling and extraction methods with possible application to pear thrips (Thysanoptera: Thripidae)

    Science.gov (United States)

    John E. Bater

    1991-01-01

    Techniques are described for the sampling and extraction of microarthropods from soil and the potential of these methods to extract the larval stages of the pear thrips, Taeniothrips inconsequens (Uzel), from soil cores taken in sugar maple stands. Also described is a design for an emergence trap that could be used to estimate adult thrips...

  17. Determination of chlormequat in pears by liquid chromatography/mass spectrometry

    NARCIS (Netherlands)

    Mol, H.G.J.; Dam, R.C.J. van; Vreeken, R.J.; Steijger, O.M.

    2000-01-01

    A straightforward and reliable method was developed for the determination of chlormequat in pears by liquid chromatography/mass spectrometry (LC/MS). Water and methanol were compared as extraction solvents. Because no significant differences in extraction efficiency or repeatability were found,

  18. How to prevent ripening blockage in 1-MCP-treated 'Conference' pears.

    Science.gov (United States)

    Chiriboga, Maria-Angeles; Schotsmans, Wendy C; Larrigaudière, Christian; Dupille, Eve; Recasens, Inmaculada

    2011-08-15

    Some European pear varieties treated with 1-methylcyclopropene (1-MCP) often remain 'evergreen', meaning that their ripening process is blocked and does not resume after removal from cold storage. In this work this was confirmed also to be the case in 'Conference' pears. To reverse the blockage of ripening 1-MCP treatments combined with external exogenous ethylene were tested. 1-MCP treatment of 'Conference' pears is very effective in delaying ripening and, more specifically, softening. The same 1-MCP concentration in different experimental years caused a different response. The higher dose of 1-MCP (600 nL L⁻¹) always resulted in irreversible blockage of ripening, whereas the behaviour of fruit receiving a lower dose (300 nL L⁻¹) depended on the year, and this did not depend on maturity at harvest or on storage conditions. Simultaneous exposure to 1-MCP and exogenous ethylene significantly affected fruit ripening, allowing significant softening to occur but at a lower rate compared with control fruit. The application of exogenous ethylene and 1-MCP simultaneously after harvest permitted restoration of the ripening process after storage in 'Conference' pears, extending the possibility of marketing and consumption. Copyright © 2011 Society of Chemical Industry.

  19. Improved irrigation scheduling for pear-jujube trees based on trunk ...

    African Journals Online (AJOL)

    A suitable indicator for scheduling pear-jujube (Ziziphus jujuba Mill.) irrigation in China was developed based on trunk diameter fluctuations (TDF). Parameters derived from TDF responses to variations in soil matrix potential (Ψsoil) were compared under deficit and well irrigation. Maximum daily shrinkage (MDS) increased ...

  20. Hourly laying patterns of the Pearly-eyed Thrasher (Margarops fuscatus) in Puerto Rico

    Science.gov (United States)

    Wayne Arendt

    2011-01-01

    Temporal aspects of egg deposition are important factors governing avian reproductive success. I report hourly egg-laying patterns of the Pearly-eyed Thrasher (Margarops fuscatus) in the Luquillo Experimental Forest in northeastern Puerto Rico during 1979–2000. Initiatory eggs were laid by early morning (median 5 0642 hrs, AST) and almost half of the eggs were laid by...

  1. Sources of Neofabraea spp. and Cadophora spp. in Dutch apple and pear orchards

    NARCIS (Netherlands)

    Köhl, J.; Wenneker, M.; Haas, de B.H.; Anbergen, R.; Lombaers-van der Plas, C.H.; Kastelein, P.

    2015-01-01

    Post-harvest diseases of apple and pear result in significant economic losses during long storage. Pathogens causing quiescent infections in the orchard leading to late post-harvest losses in The Netherlands are Neofabraea alba (Lenticel spot disease), N. perennans (Bull eye rot), Neonectria

  2. High prevalence of pollinosis symptoms among the farmers cultivating Japanese pears.

    Science.gov (United States)

    Hayashi, S; Teranishi, H; Shimooka, Y; Yamada, N

    2007-01-01

    In a district of Japanese pear cultivators, a questionnaire survey and an IgE antibody survey were conducted on the pollinosis. A high prevalence of 36.3 percent of the farmers complained of pollinosis symptoms. By the IgE antibody survey, the symptoms were found to be related to the airborne pollens in the orchard.

  3. Overhead work and shoulder-neck pain in orchard farmers harvesting pears and apples.

    Science.gov (United States)

    Sakakibara, H; Miyao, M; Kondo, T; Yamada, S

    1995-04-01

    The effects of overhead work were studied by comparing orchard farmers' musculoskeletal symptoms while bagging pears with those same symptoms while bagging apples. The subjects were 52 Japanese female farmers, who were examined twice an evening in late June for bagging pears, and during another evening of late July for bagging apples, when each task had been almost finished. They were questioned about musculoskeletal complaints of stiffness and pain during each job, and examined for muscle tenderness and pain from joint movement. Arm elevation angles during the work were measured for each type of bagging. The prevalence of stiffness and pain in the neck and shoulder, muscle tenderness in the shoulder regions, and pain in neck motion were found to be significantly higher when bagging pears than apples. Musculoskeletal symptoms of parts other than the neck and shoulder did not differ between the two types of bagging. The working posture of elevating the arm more than 90 degrees was assumed to account for 75% of the time bagging pears, against 40% for bagging apples. Overhead work requiring arm elevation and head extension was considered to be closely related with shoulder-neck disorders among farmers.

  4. Effects of major geometric variations between intracavitary applications on pear-shaped isodose dimension in cancer of the cervix

    International Nuclear Information System (INIS)

    Kim, R. Y.

    1996-01-01

    PURPOSE: The basic principal of intracavitary brachytherapy for cancer of the cervix is based on specific loading rules to achieve a pear-shaped isodose distribution centered around the cervix. Recently, ICRU Report 38 recommends a dose reference volume for reporting. Our previous studies have confirmed that there is considerable variations of geometry between applications. This study is to evaluate the effect of major geometric variations on pear-shaped isodose dimension in manual afterloading low-dose-rate system. MATERIAL AND METHODS: One hundred orthogonal films of 50 patients with cancer of the cervix (2 applications/patient) were reviewed for comparative measurements of geometric variations between applications. Major geometric variations were found for 13 patients in lengths of tandem, 7 patients in colpostats separation and 16 patients in vaginal packing. The direct measurement of these geometric variations were compared with the three-dimensional measurement of the pear-shaped isodose enclosed by the point A between the two applications. RESULTS: The geometric variations in the width of colpostats separation and length of tandem were directly related to the width and height of the pear-shaped isodose dimension. The geometric relationship between the colpostats and distal tandem had an important effect on the thickness of the pear-shape. In optimization of poor geometry for rectum or bladder wall, high dose volume centered around the cervix is reduced without changing the overall pear-shaped volume due to changing configuration of the pear-shaped isodose. In our selected patients with two applications, variations in vaginal packing had no direct effect on the width and thickness of the pear-shape due to other variables. CONCLUSION: Major geometric variations between applications greatly affect the dimension of the pear-shaped isodose distribution. Optimization of poor geometry is quite limited without compromising the high-dose volume centered around the

  5. Investigation of Pear Drying Performance by Different Methods and Regression of Convective Heat Transfer Coefficient with Support Vector Machine

    Directory of Open Access Journals (Sweden)

    Mehmet Das

    2018-01-01

    Full Text Available In this study, an air heated solar collector (AHSC dryer was designed to determine the drying characteristics of the pear. Flat pear slices of 10 mm thickness were used in the experiments. The pears were dried both in the AHSC dryer and under the sun. Panel glass temperature, panel floor temperature, panel inlet temperature, panel outlet temperature, drying cabinet inlet temperature, drying cabinet outlet temperature, drying cabinet temperature, drying cabinet moisture, solar radiation, pear internal temperature, air velocity and mass loss of pear were measured at 30 min intervals. Experiments were carried out during the periods of June 2017 in Elazig, Turkey. The experiments started at 8:00 a.m. and continued till 18:00. The experiments were continued until the weight changes in the pear slices stopped. Wet basis moisture content (MCw, dry basis moisture content (MCd, adjustable moisture ratio (MR, drying rate (DR, and convective heat transfer coefficient (hc were calculated with both in the AHSC dryer and the open sun drying experiment data. It was found that the values of hc in both drying systems with a range 12.4 and 20.8 W/m2 °C. Three different kernel models were used in the support vector machine (SVM regression to construct the predictive model of the calculated hc values for both systems. The mean absolute error (MAE, root mean squared error (RMSE, relative absolute error (RAE and root relative absolute error (RRAE analysis were performed to indicate the predictive model’s accuracy. As a result, the rate of drying of the pear was examined for both systems and it was observed that the pear had dried earlier in the AHSC drying system. A predictive model was obtained using the SVM regression for the calculated hc values for the pear in the AHSC drying system. The normalized polynomial kernel was determined as the best kernel model in SVM for estimating the hc values.

  6. The identification of two Trypanosoma cruzi I genotypes from domestic and sylvatic transmission cycles in Colombia based on a single polymerase chain reaction amplification of the spliced-leader intergenic region

    Directory of Open Access Journals (Sweden)

    Lina Marcela Villa

    2013-11-01

    Full Text Available A single polymerase chain reaction (PCR reaction targeting the spliced-leader intergenic region of Trypanosoma cruzi I was standardised by amplifying a 231 bp fragment in domestic (TcIDOM strains or clones and 450 and 550 bp fragments in sylvatic strains or clones. This reaction was validated using 44 blind coded samples and 184 non-coded T. cruzi I clones isolated from sylvatic triatomines and the correspondence between the amplified fragments and their domestic or sylvatic origin was determined. Six of the nine strains isolated from acute cases suspected of oral infection had the sylvatic T. cruzi I profile. These results confirmed that the sylvatic T. cruzi I genotype is linked to cases of oral Chagas disease in Colombia. We therefore propose the use of this novel PCR reaction in strains or clones previously characterised as T. cruzi I to distinguish TcIDOMfrom sylvatic genotypes in studies of transmission dynamics, including the verification of population selection within hosts or detection of the frequency of mixed infections by both T. cruzi I genotypes in Colombia.

  7. Development of a nested polymerase chain reaction for amplification of a sequence of the p57 gene of Renibacterium salmoninarum that provides a highly sensitive method for detection of the bacterium in salmonid kidney

    Science.gov (United States)

    Chase, D.M.; Pascho, R.J.

    1998-01-01

    Nucleic acid-based assays have shown promise for diagnosing Renibacterium salmoninarum in tissues and body fluids of salmonids. DeVelopment of a nested polymerase chain reaction (PCR) method to detect a 320 bp DNA segment of the gene encoding the p57 protein of R. salmoninarum is described. Whereas a conventional PCR for a 383 bp segment of the p57 gene reliably detected 1000 R. salmoninarum cells per reaction in kidney tissue, the nested PCR detected as few as 10 R. salmoninarum per reaction in kidney tissue. Two DNA extraction methods for the nested PCR were compared and the correlation between replicate samples was generally higher in samples extracted by the QIAamp system compared with those extracted by the phenol/chloroform method. The specificity of the nested PCR was confirmed by testing DNA extracts of common bacterial fish pathogens and a panel of bacterial species reported to cause false-positive reactions in the enzyme-linked immunosorbent assay (ELISA) and the fluorescent antibody test (FAT) for R. salmoninarum. Kidney samples from 74 naturally infected chinook Salmon were examined by the nested PCR, the ELISA, and the FAT, and the detected prevalences of R. salmoninarum were 61, 47, and 43%, respectively.

  8. PEAR SHOOT SAWFLY (JANUS COMPRESSUS FABRICIUS – LIFE CYCLE AND BIOLOGICAL AND MORPHOLOGICAL CHARACTERISTIC

    Directory of Open Access Journals (Sweden)

    Tihomir Validžić

    2014-06-01

    Full Text Available The aim of the thesis was to investigate life cycle, biological and morphological characteristics of pear shoot sawfly (Janus compressus Fabricius, Hymenoptera Cephidae, furthermore to identify natural enemies in order to protect pear from this pest. The trial was conducted in the period of three years: 2010, 2011 and 2012 in pear orchards at five localities. Monitoring of adult sawfly was done by yellow sticky traps. Laboratory research was done at the Faculty of Agriculture, Department of Plant Protection, Section of Entomology and Nematology. In this study, pear shoot sawfly in Eastern Slavonia occurred in the period of four weeks, starting from the third decade of April with the peak population at the beginning of the May. Adults flight is the most intensive during warm and sunny days, when temperatures are above 14°C. Adult sawflies are characterized by elongated body and antennae, usually 7-12 mm long and sexual dimorphism is present. Pest is univoltine. Basic colour of adult sawfly is black. Antennae are moniliform and consist of 20 (male - 22 (female segments. Females have red or dark red colored abdomen, while males have yellow or orange one. Eggs are cylindrically shaped, 0.8-1.0 mm long. Female lays approximately 30 eggs. Embryonic development of pear shoot sawfly eggs lasts from 11 to 14 days. Larvae are 8-10 mm long, white or pale yellow. Larvae molt three times. Pear shoot sawfly larvae were parasitized by insects from Hymenoptera order, from five identified and one unidentified genera. Level of parasitism by genera is as follows: Eurytoma sp. (Hymenoptera: Eurytomidae – 9.83%, Tetrastichus sp. (Hymenoptera: Eulophidae – 2.01%, Eupelmus sp. (Hymenoptera: Eupelmidae – 1.66%, Pteromalus sp. (Hymenoptera: Pteromalidae – 0.55%, Ichneumonida sp. (Hymenoptera: Pimplinae – 0.35% and unidentified genera – 0.62%. Plant parasitic species Metopoplax origani (Hemiptera: Lygaeidae was found in 1.80% of analyzed shoots. Larvae were

  9. Extraction and determination of polyphenols and betalain pigments in the Moroccan Prickly pear fruits (Opuntia ficus indica

    Directory of Open Access Journals (Sweden)

    Omar Khatabi

    2016-09-01

    This study has permitted us equally to value betalain pigments extracted from fruity juice. These are the betalains present in the epidermis and the pulp of the prickly pear confers on it its color varying from yellow to purple. Results show that yellow and red prickly pears contain imported strengths in betalains. Our work shows that the red prickly pear contains betaxanthin pigments in excess of the indicaxanthin that permits to valorize human’s potential spring of genuine colorings. Betalains and polyphenols are antioxidants that contribute to nutritional prickly pears’ quality and to their products of transformation.

  10. Alternative Chemical Amplification Methods for Peroxy Radical Detection

    Science.gov (United States)

    Wood, E. C. D.

    2014-12-01

    Peroxy radicals (HO2, CH3O2, etc.) are commonly detected by the chemical amplification technique, in which ambient air is mixed with high concentrations of CO and NO, initiating a chain reaction that produces 30 - 200 NO2 molecules per sampled peroxy radical. The NO2 is then measured by one of several techniques. With the exception of CIMS-based techniques, the chemical amplification method has undergone only incremental improvements since it was first introduced in 1982. The disadvantages of the technique include the need to use high concentrations of CO and the greatly reduced sensitivity of the amplification chain length in the presence of water vapor. We present a new chemical amplification scheme in which either ethane or acetaldehyde is used in place of CO, with the NO2 product detected using Cavity Attenuated Phase Shift spectroscopy (CAPS). Under dry conditions, the amplification factor of the alternative amplifiers are approximately six times lower than the CO-based amplifier. The relative humidity "penalty" is not as severe, however, such that at typical ambient relative humidity (RH) values the amplification factor is within a factor of three of the CO-based amplifier. Combined with the NO2 sensitivity of CAPS and a dual-channel design, the detection limit of the ethane amplifier is less than 2 ppt (1 minute average, signal-to-noise ratio 2). The advantages of these alternative chemical amplification schemes are improved safety, a reduced RH correction, and increased sensitivity to organic peroxy radicals relative to HO2.

  11. Shelf-life extension and quality improvement of minimally processed pear by combination treatments with irradiation

    International Nuclear Information System (INIS)

    Swailam, H.M.; Hammad, A.A.; Serag, M.S.; Mansour, F.A.; Abu El-Nour, S.A.

    2007-01-01

    Fresh-cut pears are not yet commercially available in Egypt. Thus, an attempt has been done to produce this minimally processed fruit manually at the laboratory. Fifteen samples of this manually prepared fresh-produce were evaluated for their microbiological quality. Total aerobic bacterial counts (TAPC) ranged from 7.5 x 10 to 3.5 x 10 cfu g-1, lactic acid bacteria (LAB) ranged from less than 10 to 3.2 x 10 cfu g-1; total mould and yeast counts (TM & Y) ranged from less than 10 to 5.3 x 10 cfu g-1 indicating good quality from the view point of microbiological population. Coliform bacteria and Escherichia coli were found in only 2 samples at levels of 20 to 43 and greater than 3 to 9 most probable numbers per gram (MPN g-1), respectively. On the other hand, Staphylococcus aureus was found in also 2 samples at levels of 102. Aeromonas hydrohila, Listeria monocytogenes and Salmonella spp., were not found in any of the samples. Fresh-cut pear s were dipped in water containing 2% ascorbic acid and 1% calcium lactate. The dipped fresh-cut pears were exposed to 1, 2 and 3 kGy. Dipping fresh-cut pears in 2% ascorbic acid plus 1% calcium lactate prevented browning and enhanced firmness during refrigeration storage. Irradiation dose of 2 kGy was the optimum, since it reduced TAPC by 99.58% and LAB and TM and Y to un-detectable level. These combined treatments extend the shelf life of fresh-cut pears without affecting their chemical, physical and sensorial quality attributes

  12. CDK4 amplification predicts recurrence of well-differentiated liposarcoma of the abdomen.

    Directory of Open Access Journals (Sweden)

    Sanghoon Lee

    Full Text Available The absence of CDK4 amplification in liposarcomas is associated with favorable prognosis. We aimed to identify the factors associated with tumor recurrence in patients with well-differentiated (WD and dedifferentiated (DD liposarcomas.From 2000 to 2010, surgical resections for 101 WD and DD liposarcomas were performed. Cases in which complete surgical resections with curative intent were carried out were selected. MDM2 and CDK4 gene amplification were analyzed by quantitative real-time polymerase chain reaction (Q-PCR.There were 31 WD and 17 DD liposarcomas. Locoregional recurrence was observed in 11 WD and 3 DD liposarcomas. WD liposarcomas showed better patient survival compared to DD liposarcomas (P<0.05. Q-PCR analysis of the liposarcomas revealed the presence of CDK4 amplification in 44 cases (91.7% and MDM2 amplification in 46 cases (95.8%. WD liposarcomas with recurrence after surgical resection had significantly higher levels of CDK4 amplification compared to those without recurrence (P = 0.041. High level of CDK4 amplification (cases with CDK4 amplification higher than the median 7.54 was associated with poor recurrence-free survival compared to low CDK4 amplification in both univariate (P = 0.012 and multivariate analyses (P = 0.020.Level of CDK4 amplification determined by Q-PCR was associated with the recurrence of WD liposarcomas after surgical resection.

  13. An evaluation of multiple annealing and looping based genome amplification using a synthetic bacterial community

    KAUST Repository

    Wang, Yong

    2016-02-23

    The low biomass in environmental samples is a major challenge for microbial metagenomic studies. The amplification of a genomic DNA was frequently applied to meeting the minimum requirement of the DNA for a high-throughput next-generation-sequencing technology. Using a synthetic bacterial community, the amplification efficiency of the Multiple Annealing and Looping Based Amplification Cycles (MALBAC) kit that is originally developed to amplify the single-cell genomic DNA of mammalian organisms is examined. The DNA template of 10 pg in each reaction of the MALBAC amplification may generate enough DNA for Illumina sequencing. Using 10 pg and 100 pg templates for each reaction set, the MALBAC kit shows a stable and homogeneous amplification as indicated by the highly consistent coverage of the reads from the two amplified samples on the contigs assembled by the original unamplified sample. Although GenomePlex whole genome amplification kit allows one to generate enough DNA using 100 pg of template in each reaction, the minority of the mixed bacterial species is not linearly amplified. For both of the kits, the GC-rich regions of the genomic DNA are not efficiently amplified as suggested by the low coverage of the contigs with the high GC content. The high efficiency of the MALBAC kit is supported for the amplification of environmental microbial DNA samples, and the concerns on its application are also raised to bacterial species with the high GC content.

  14. Change in chemical constituents and free radical-scavenging activity during Pear (Pyrus pyrifolia) cultivar fruit development.

    Science.gov (United States)

    Cho, Jeong-Yong; Lee, Sang-Hyun; Kim, Eun Hee; Yun, Hae Rim; Jeong, Hang Yeon; Lee, Yu Geon; Kim, Wol-Soo; Moon, Jae-Hak

    2015-01-01

    Changes in chemical constituent contents and DPPH radical-scavenging activity in fruits of pear (Pyrus pyrifolia) cultivars during the development were investigated. The fruits of seven cultivars (cv. Niitaka, Chuhwangbae, Wonhwang, Hwangkeumbae, Hwasan, Manpungbae, and Imamuraaki) were collected at 15-day intervals after day 20 of florescence. Vitamins (ascorbic acid and α-tocopherol), arbutin, chlorogenic acid, malaxinic acid, total caffeic acid, total flavonoids, and total phenolics were the highest in immature pear fruit on day 20 after florescence among samples at different growth stages. All of these compounds decreased gradually in the fruit during the development. Immature pear fruit on day 35 or 50 after florescence exhibited higher free radical-scavenging activity than that at other times, although activities were slightly different among cultivars. The chemical constituent contents and free radical-scavenging activity were largely different among immature fruits of the pear cultivars, but small differences were observed when they matured.

  15. Studies on the Paint Forming Properties of Avocado(Persea Americana) and African Pear (Dacryodes Edulis) Seed Oils.

    OpenAIRE

    Otaigbe, J.O.E; Oriji, O.G; Ekerenam, G.E

    2016-01-01

    Avocado(Persea Americana) and African Pear (Dacryodes edulis) seed oils were investigated for their suitability as base materials for oil paint production. Soxhlet extraction of the oils from the powdered seeds using n-hexane gave 3.63% and 10.40% yields for Avocado and African Pear respectively. Proximate analysis and chemical characterization of the seed oils were carried out using standard procedures according to the American Oil Chemist Society (AOCS) and the American Society for Testing ...

  16. Whole genome amplification in preimplantation genetic diagnosis*

    Science.gov (United States)

    Zheng, Ying-ming; Wang, Ning; Li, Lei; Jin, Fan

    2011-01-01

    Preimplantation genetic diagnosis (PGD) refers to a procedure for genetically analyzing embryos prior to implantation, improving the chance of conception for patients at high risk of transmitting specific inherited disorders. This method has been widely used for a large number of genetic disorders since the first successful application in the early 1990s. Polymerase chain reaction (PCR) and fluorescent in situ hybridization (FISH) are the two main methods in PGD, but there are some inevitable shortcomings limiting the scope of genetic diagnosis. Fortunately, different whole genome amplification (WGA) techniques have been developed to overcome these problems. Sufficient DNA can be amplified and multiple tasks which need abundant DNA can be performed. Moreover, WGA products can be analyzed as a template for multi-loci and multi-gene during the subsequent DNA analysis. In this review, we will focus on the currently available WGA techniques and their applications, as well as the new technical trends from WGA products. PMID:21194180

  17. Genome position and gene amplification

    Czech Academy of Sciences Publication Activity Database

    Jirsová, Pavla; Snijders, A.M.; Kwek, S.; Roydasgupta, R.; Fridlyand, J.; Tokuyasu, T.; Pinkel, D.; Albertson, D. G.

    2007-01-01

    Roč. 8, č. 6 (2007), r120 ISSN 1474-760X Institutional research plan: CEZ:AV0Z50040507; CEZ:AV0Z50040702 Keywords : gene amplification * array comparative genomic hybridization * oncogene Subject RIV: BO - Biophysics Impact factor: 6.589, year: 2007

  18. Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 Triggered Isothermal Amplification for Site-Specific Nucleic Acid Detection.

    Science.gov (United States)

    Huang, Mengqi; Zhou, Xiaoming; Wang, Huiying; Xing, Da

    2018-02-06

    A novel CRISPR/Cas9 triggered isothermal exponential amplification reaction (CAS-EXPAR) strategy based on CRISPR/Cas9 cleavage and nicking endonuclease (NEase) mediated nucleic acids amplification was developed for rapid and site-specific nucleic acid detection. CAS-EXPAR was primed by the target DNA fragment produced by cleavage of CRISPR/Cas9, and the amplification reaction performed cyclically to generate a large number of DNA replicates which were detected using a real-time fluorescence monitoring method. This strategy that combines the advantages of CRISPR/Cas9 and exponential amplification showed high specificity as well as rapid amplification kinetics. Unlike conventional nucleic acids amplification reactions, CAS-EXPAR does not require exogenous primers, which often cause target-independent amplification. Instead, primers are first generated by Cas9/sgRNA directed site-specific cleavage of target and accumulated during the reaction. It was demonstrated this strategy gave a detection limit of 0.82 amol and showed excellent specificity in discriminating single-base mismatch. Moreover, the applicability of this method to detect DNA methylation and L. monocytogenes total RNA was also verified. Therefore, CAS-EXPAR may provide a new paradigm for efficient nucleic acid amplification and hold the potential for molecular diagnostic applications.

  19. Direct amplification of casework bloodstains using the Promega PowerPlex(®) 21 PCR amplification system.

    Science.gov (United States)

    Gray, Kerryn; Crowle, Damian; Scott, Pam

    2014-09-01

    A significant number of evidence items submitted to Forensic Science Service Tasmania (FSST) are blood swabs or bloodstained items. Samples from these items routinely undergo phenol:chloroform:isoamyl alcohol organic extraction and quantitative Polymerase Chain Reaction (qPCR) testing prior to PowerPlex(®) 21 amplification. This multi-step process has significant cost and timeframe implications in a fiscal climate of tightening government budgets, pressure towards improved operating efficiencies, and an increasing emphasis on rapid techniques better supporting intelligence-led policing. Direct amplification of blood and buccal cells on cloth and Whatman FTA™ card with PowerPlex(®) 21 has already been successfully implemented for reference samples, eliminating the requirement for sample pre-treatment. Scope for expanding this method to include less pristine casework blood swabs and samples from bloodstained items was explored in an endeavour to eliminate lengthy DNA extraction, purification and qPCR steps for a wider subset of samples. Blood was deposited onto a range of substrates including those historically found to inhibit STR amplification. Samples were collected with micro-punch, micro-swab, or both. The potential for further fiscal savings via reduced volume amplifications was assessed by amplifying all samples at full and reduced volume (25 and 13μL). Overall success rate data showed 80% of samples yielded a complete profile at reduced volume, compared to 78% at full volume. Particularly high success rates were observed for the blood on fabric/textile category with 100% of micro-punch samples yielding complete profiles at reduced volume and 85% at full volume. Following the success of this trial, direct amplification of suitable casework blood samples has been implemented at reduced volume. Significant benefits have been experienced, most noticeably where results from crucial items have been provided to police investigators prior to interview of

  20. Adsorption-desorption isotherms and heat of sorption of prickly pear fruit (Opuntia ficus indica)

    International Nuclear Information System (INIS)

    Lahsasni, S.; Kouhila, M.; Mahrouz, M.

    2004-01-01

    The equilibrium moisture contents were determined for prickly pear fruit using the gravimetric static method at t=30, 40 and 50 deg. C over a range of relative humidities from 0.05 to 0.9. The sorption curves of prickly pear fruit decreased with increase in temperature at constant relative humidity. The hysteresis effect was observed. The GAB, modified Halsey, modified Chung-Pfost, modified Oswin and modified Henderson models were tested to fit the experimental data. The GAB model was found to be the most suitable for describing the sorption curves. The monolayer moisture content values for the sorption at different temperatures are calculated using a modified BET equation. The isosteric heats of desorption and adsorption of water were determined from the equilibrium data at different temperatures

  1. Adsorption-desorption isotherms and heat of sorption of prickly pear fruit (Opuntia ficus indica)

    Energy Technology Data Exchange (ETDEWEB)

    Lahsasni, S.; Kouhila, M. E-mail: kouhila@hotmail.com; Mahrouz, M

    2004-01-01

    The equilibrium moisture contents were determined for prickly pear fruit using the gravimetric static method at t=30, 40 and 50 deg. C over a range of relative humidities from 0.05 to 0.9. The sorption curves of prickly pear fruit decreased with increase in temperature at constant relative humidity. The hysteresis effect was observed. The GAB, modified Halsey, modified Chung-Pfost, modified Oswin and modified Henderson models were tested to fit the experimental data. The GAB model was found to be the most suitable for describing the sorption curves. The monolayer moisture content values for the sorption at different temperatures are calculated using a modified BET equation. The isosteric heats of desorption and adsorption of water were determined from the equilibrium data at different temperatures.

  2. Performance of 'Rocha' and 'Santa Maria' pears as affected by planting density

    Directory of Open Access Journals (Sweden)

    Mateus da Silveira Pasa

    2015-02-01

    Full Text Available The objective of this work was to evaluate the performance of 'Rocha' and 'Santa Maria' pears at two planting densities. The experiment was carried out during the 2011/2012, 2012/2013, and 2013/2014 growing seasons, in one-year-old orchards (2011/2012 of 'Rocha' and 'Santa Maria' pears, trained in a central-leader system and planted in two densities (2,000 and 4,000 trees per hectare. The assessed parameters were: production per hectare, production per tree, yield efficiency, number of fruit per tree, average fruit weight, trunk diameter increment, fruit firmness, and soluble solid contents. The cumulative yield of 'Rocha' is greater at the higher planting density, whereas the yield efficiency of 'Santa Maria' increases at the lower planting density, as the trees get more mature. Trunk diameter of 'Rocha' also increases at the lower planting density. However, fruit quality parameters in both cultivars are little affected by planting density.

  3. Measurement of indicator genes using global complementary DNA (cDNA) amplification, by polyadenylic acid reverse transcriptase polymerase chain reaction (poly A RT-PCR): A feasibility study using paired samples from tissue and ductal juice in patients undergoing pancreatoduodenectomy.

    Science.gov (United States)

    Sanyal, Sudip; Siriwardena, Ajith K; Byers, Richard

    2018-06-01

    The aim of this study is to compare gene expression profiles in RNA isolated from pancreatic ductal juice with the RNA expression profiles of the same genes from matched intra-operative tissue samples from pancreatic tumours. Intra-operative sampling of pancreatic juice and collection of matched tissue samples was undertaken in patients undergoing pancreatoduodenectomy for clinically suspected pancreatic cancer and a precursor lesion, main-duct intraductal papillary mucinous neoplasm. RNA was isolated and Poly A PCR was used to globally amplify the RNA. Real-time polymerase chain reaction (RT-PCR) was used to measure expression levels of 17 genes selected from microarray studies. Spearman's rank correlation test was used to examine the relationship of gene expression between pancreatic juice and tissue. The study was approved by Regional Ethics Committee. Mesothelin (MSLN) showed significant correlation (p cDNA using poly A PCR is technically feasible. Application of the technique to non-invasively obtained pancreatic juice during endoscopic assessment of tumours and the use of gene arrays of cancer indicator genes are the next steps in development of this technique. Copyright © 2018 IAP and EPC. Published by Elsevier B.V. All rights reserved.

  4. Effect of irradiation and cold storage on the physiological characteristics of the pear fruits

    International Nuclear Information System (INIS)

    Mahmoud, A.A.; Roushdy, H.M.; El-Latif, F.A.; Taha, F.

    1988-01-01

    With the aim of extending shelf life of fruits and the effects of gamma irradiation at the 0, 1, 2, 3 and 4 KGy dose levels, the present investigation has been undertaken to illustrate, the possible application of gamma irradiation processing to prolong the storage period of mature pear fruit. Significant differences had been found between irradiated and non-irradiated fruits regarding total soluble solids, weight loss, decay, firmness, titrable acidity and moisture content

  5. Thin layer convective solar drying and mathematical modeling of prickly pear peel (Opuntia ficus indica)

    International Nuclear Information System (INIS)

    Lahsasni, Siham; Kouhila, Mohammed; Mahrouz, Mostafa; Idlimam, Ali; Jamali, Abdelkrim

    2004-01-01

    This paper presents the thin layer convective solar drying and mathematical modeling of prickly pear peel. For these purposes, an indirect forced convection solar dryer consisting of a solar air collector, an auxiliary heater, a circulation fan and a drying cabinet is used for drying experiments. Moreover, the prickly pear peel is sufficiently dried in the ranges of 32 to 36 deg. C of ambient air temperature, 50 to 60 deg. C of drying air temperature, 23 to 34% of relative humidity, 0.0277 to 0.0833 m 3 /s of drying air flow rate and 200 to 950 W/m 2 of daily solar radiation. The experimental drying curves show only a falling drying rate period. The main factor in controlling the drying rate was found to be the drying air temperature. The drying rate equation is determined empirically from the characteristic drying curve. Also, the experimental drying curves obtained were fitted to a number of mathematical models. The Midilli-Kucuk drying model was found to satisfactorily describe the solar drying curves of prickly pear peel with a correlation coefficient (r) of 0.9998 and chi-square (χ 2 ) of 4.6572 10 -5

  6. Thin layer convective solar drying and mathematical modeling of prickly pear peel (Opuntia ficus indica)

    Energy Technology Data Exchange (ETDEWEB)

    Lahsasni, S.; Mahrouz, M. [Unite de Chimie Agroalimentaire (LCOA), Faculte des Sciences Semlalia, Marrakech (Morocco); Kouhila, M.; Idlimam, A.; Jamali, A. [Ecole Normale Superieure, Marrakech (Morocco). Lab. d' Energie Solaire et Plantes Aromatiques et Medicinales

    2004-02-01

    This paper presents the thin layer convective solar drying and mathematical modeling of prickly pear peel. For these purposes, an indirect forced convection solar dryer consisting of a solar air collector, an auxiliary heater, a circulation fan and a drying cabinet is used for drying experiments. Moreover, the prickly pear peel is sufficiently dried in the ranges of 32 to 36 {sup o} C of ambient air temperature, 50 to 60 {sup o}C of drying air temperature, 23 to 34% of relative humidity, 0.0277 to 0.0833 m{sup 3}/s of drying air flow rate and 200 to 950 W/m{sup 2} of daily solar radiation. The experimental drying curves show only a falling drying rate period. The main factor in controlling the drying rate was found to be the drying air temperature. The drying rate equation is determined empirically from the characteristic drying curve. Also, the experimental drying curves obtained were fitted to a number of mathematical models. The Midilli-Kucuk drying model was found to satisfactorily describe the solar drying curves of prickly pear peel with a correlation coefficient (r) of 0.9998 and chi-square ({chi}{sup 2}) of 4.6572 10{sup -5}. (Author)

  7. Use of Red Cactus Pear (Opuntia ficus-indica Encapsulated Powder to Pigment Extruded Cereal

    Directory of Open Access Journals (Sweden)

    Martha G. Ruiz-Gutiérrez

    2017-01-01

    Full Text Available Encapsulated powder of the red cactus pear is a potential natural dye for the food industry and a known antioxidant. Although the use of this powder is possible, it is not clear how it alters food properties, thus ensuing commercial acceptability. The aim of this study was to evaluate the effect of encapsulated powder of the red cactus pear on the physicochemical properties of extruded cereals. The powder was mixed (2.5, 5.0, and 7.5% w/w with maize grits and extruded (mix moisture 22%, temperature 100°C, and screw speed 325 rpm. The physical, chemical, and sensory characteristics of the extruded cereal were evaluated; extruded cereal without encapsulated powder was used as a control. All cereal extrudates pigmented with the encapsulated powder showed statistically significant differences (P<0.05 in expansion, water absorption, color, density, and texture compared to the control. The encapsulated powder had a positive effect on expansion and water absorption indices, as well as color parameters, but a negative effect on density and texture. Extruded cereal properties were significantly (P<0.05 correlated. Sensorially, consumers accepted the extruded cereal with a lower red cactus pear powder content (2.5% w/w, because this presented characteristics similar to extruded cereal lacking pigment.

  8. Aureobasidium pullulansas a biocontrol agent of blue mold in "Rocha" pear.

    Science.gov (United States)

    Ferreira-Pinto, M M; Moura-Guedes, M C; Barreiro, M G; Pais, I; Santos, M R; Silva, M J

    2006-01-01

    The blue mold of "Rocha" pear caused by Penicillium expansum is an important postharvest disease which is adequately controlled by application of synthetic fungicides. In recent years, strategies like biological control have been considered a desirable alternative to chemicals. Several studies have demonstrated the potential of the yeast-like fungus Aureobasidium pullulans for control of postharvest decay of pear. A Portuguese isolate of Aureobasidium pullulans was characterized and evaluated for its activity in reducing postharvest blue mold decay of "Rocha" pear caused by Penicillium expansum. Study of optimal conditions for antagonist growth was carried out in six different culture media. The effect of four maturity stages of fruits in the development of A. pullulans was also studied. Biocontrol studies were performed with two concentrations of the antagonist (3 x 10(8) and 4 x 10(9) CFU/ml). A. pullulans growth was significantly different (P Corn Meal Agar (CMA) and Potato Dextrose Agar (PDA) media which contains the higher concentration of glucose (20 mg/l). Medium resulted from fruits of the first harvest date presented lower colony diameter. Inoculation of A. pullulans at 3 x 10(8) and 4 x 10(9) CFU/ml reduced the incidence of the disease by 23 and 63%, and reduced the lesion diameter by 36 and 46%, respectively.

  9. ‘PACKHAM’S TRIUMPH’ PEAR RESPONSE TO 1-METHYLCYCLOPROPENE AND NITRIC OXIDE TREATMENTS

    Directory of Open Access Journals (Sweden)

    MARCOS VINÍCIUS HENDGES

    2016-01-01

    Full Text Available This study aimed at assessing the effect of 1-methylcyclopropene (1-MCP, gas-nitric oxide (NO and sodium nitroprusside (SNP on ripening of ‘Packham’s Triumph’ pears. The treatments consisted of T1 control; T2 300 ppb 1-MCP; T3 1 mM SNP; T4 10 ppm NO; and T5 20 ppm NO. The fruit treated with 1-MCP showed significantly higher values for flesh firmness, texture, and peel green color, besides lower respiratory rates and ethylene production. On the other hand, NO and SNP treatments did not reduce fruit respiratory rate and ethylene production. Flesh firmness and textural features were maintained by treating fruit with 20 ppm NO after leaving chambers. Treatments using 1 mM SNP and 20 ppm NO kept peel green color (higher hue angle when compared to control, without decreasing fruit yellowing during the shelf life. The application of 300 ppb 1-MCP prevented buttery texture and yellowing in 'Packham’s Triumph' pear fruit during environment condition exposure. The use NO at 20 ppm kept flesh firmness during storage, however, with subsequent reduction of this variable in environmental conditions. The treatments with 1mM SNP and 20 ppm NO maintained green peel of pear fruit, even after exposure to environmental conditions, but not limiting yellowing.

  10. Signal amplification of microRNAs with modified strand displacement-based cycling probe technology.

    Science.gov (United States)

    Jia, Huning; Bu, Ying; Zou, Bingjie; Wang, Jianping; Kumar, Shalen; Pitman, Janet L; Zhou, Guohua; Song, Qinxin

    2016-10-24

    Micro ribose nucleic acids (miRNAs) play an important role in biological processes such as cell differentiation, proliferation and apoptosis. Therefore, miRNAs are potentially a powerful marker for monitoring cancer and diagnosis. Here, we present sensitive signal amplification for miRNAs based on modified cycling probe technology with strand displacement amplification. miRNA was captured by the template coupled with beads, and then the first cycle based on SDA was repeatedly extended to the nicking end, which was produced by the extension reaction of miRNA. The products generated by SDA are captured by a molecular beacon (MB), which is designed to initiate the second amplification cycle, with a similar principle to the cycling probe technology (CPT), which is based on repeated digestion of the DNA-RNA hybrid by the RNase H. After one sample enrichment and two steps of signal amplification, 0.1 pM of let-7a can be detected. The miRNA assay exhibits a great dynamic range of over 100 orders of magnitude and high specificity to clearly discriminate a single base difference in miRNA sequences. This isothermal amplification does not require any special temperature control instrument. The assay is also about signal amplification rather than template amplification, therefore minimising contamination issues. In addition, there is no need for the reverse transcription (RT) process. Thus the amplification is suitable for miRNA detection.

  11. Laser amplification in excited dielectrics

    Science.gov (United States)

    Winkler, Thomas; Haahr-Lillevang, Lasse; Sarpe, Cristian; Zielinski, Bastian; Götte, Nadine; Senftleben, Arne; Balling, Peter; Baumert, Thomas

    2018-01-01

    Wide-bandgap dielectrics such as glasses or water are transparent at visible and infrared wavelengths. This changes when they are exposed to ultrashort and highly intense laser pulses. Different interaction mechanisms lead to the appearance of various transient nonlinear optical phenomena. Using these, the optical properties of dielectrics can be controlled from the transparent to the metal-like state. Here we expand this range by a yet unexplored mechanism in excited dielectrics: amplification. In a two-colour pump-probe experiment, we show that a 400 nm femtosecond laser pulse is coherently amplified inside an excited sapphire sample on a scale of a few micrometres. Simulations strongly support the proposed two-photon stimulated emission process, which is temporally and spatially controllable. Consequently, we expect applications in all fields that demand strongly localized amplification.

  12. Measuring the amplification of attention

    OpenAIRE

    Blaser, Erik; Sperling, George; Lu, Zhong-Lin

    1999-01-01

    An ambiguous motion paradigm, in which the direction of apparent motion is determined by salience (i.e., the extent to which an area is perceived as figure versus ground), is used to assay the amplification of color by attention to color. In the red–green colored gratings used in these experiments, without attention instructions, salience depends on the chromaticity difference between colored stripes embedded in the motion sequence and the yellow background. Selective attention to red (or to ...

  13. Spheromak Impedance and Current Amplification

    International Nuclear Information System (INIS)

    Fowler, T K; Hua, D D; Stallard, B W

    2002-01-01

    It is shown that high current amplification can be achieved only by injecting helicity on the timescale for reconnection, τ REC , which determines the effective impedance of the spheromak. An approximate equation for current amplification is: dI TOR 2 /dt ∼ I 2 /τ REC - I TOR 2 /τ closed where I is the gun current, I TOR is the spheromak toroidal current and τ CLOSED is the ohmic decay time of the spheromak. Achieving high current amplification, I TOR >> I, requires τ REC CLOSED . For resistive reconnection, this requires reconnection in a cold zone feeding helicity into a hot zone. Here we propose an impedance model based on these ideas in a form that can be implemented in the Corsica-based helicity transport code. The most important feature of the model is the possibility that τ REC actually increases as the spheromak temperature increases, perhaps accounting for the ''voltage sag'' observed in some experiments, and a tendency toward a constant ratio of field to current, B ∝ I, or I TOR ∼ I. Program implications are discussed

  14. Genome-wide analysis of WRKY transcription factors in white pear (Pyrus bretschneideri) reveals evolution and patterns under drought stress.

    Science.gov (United States)

    Huang, Xiaosan; Li, Kongqing; Xu, Xiaoyong; Yao, Zhenghong; Jin, Cong; Zhang, Shaoling

    2015-12-24

    WRKY transcription factors (TFs) constitute one of the largest protein families in higher plants, and its members contain one or two conserved WRKY domains, about 60 amino acid residues with the WRKYGQK sequence followed by a C2H2 or C2HC zinc finger motif. WRKY proteins play significant roles in plant development, and in responses to biotic and abiotic stresses. Pear (Pyrus bretschneideri) is one of the most important fruit crops in the world and is frequently threatened by abiotic stress, such as drought, affecting growth, development and productivity. Although the pear genome sequence has been released, little is known about the WRKY TFs in pear, especially in respond to drought stress at the genome-wide level. We identified a total of 103 WRKY TFs in the pear genome. Based on the structural features of WRKY proteins and topology of the phylogenetic tree, the pear WRKY (PbWRKY) family was classified into seven groups (Groups 1, 2a-e, and 3). The microsyteny analysis indicated that 33 (32%) PbWRKY genes were tandemly duplicated and 57 genes (55.3%) were segmentally duplicated. RNA-seq experiment data and quantitative real-time reverse transcription PCR revealed that PbWRKY genes in different groups were induced by drought stress, and Group 2a and 3 were mainly involved in the biological pathways in response to drought stress. Furthermore, adaptive evolution analysis detected a significant positive selection for Pbr001425 in Group 3, and its expression pattern differed from that of other members in this group. The present study provides a solid foundation for further functional dissection and molecular evolution of WRKY TFs in pear, especially for improving the water-deficient resistance of pear through manipulation of the PbWRKYs.

  15. Thermally multiplexed polymerase chain reaction.

    Science.gov (United States)

    Phaneuf, Christopher R; Pak, Nikita; Saunders, D Curtis; Holst, Gregory L; Birjiniuk, Joav; Nagpal, Nikita; Culpepper, Stephen; Popler, Emily; Shane, Andi L; Jerris, Robert; Forest, Craig R

    2015-07-01

    Amplification of multiple unique genetic targets using the polymerase chain reaction (PCR) is commonly required in molecular biology laboratories. Such reactions are typically performed either serially or by multiplex PCR. Serial reactions are time consuming, and multiplex PCR, while powerful and widely used, can be prone to amplification bias, PCR drift, and primer-primer interactions. We present a new thermocycling method, termed thermal multiplexing, in which a single heat source is uniformly distributed and selectively modulated for independent temperature control of an array of PCR reactions. Thermal multiplexing allows amplification of multiple targets simultaneously-each reaction segregated and performed at optimal conditions. We demonstrate the method using a microfluidic system consisting of an infrared laser thermocycler, a polymer microchip featuring 1 μl, oil-encapsulated reactions, and closed-loop pulse-width modulation control. Heat transfer modeling is used to characterize thermal performance limitations of the system. We validate the model and perform two reactions simultaneously with widely varying annealing temperatures (48 °C and 68 °C), demonstrating excellent amplification. In addition, to demonstrate microfluidic infrared PCR using clinical specimens, we successfully amplified and detected both influenza A and B from human nasopharyngeal swabs. Thermal multiplexing is scalable and applicable to challenges such as pathogen detection where patients presenting non-specific symptoms need to be efficiently screened across a viral or bacterial panel.

  16. A cascade amplification strategy based on rolling circle amplification and hydroxylamine amplified gold nanoparticles enables chemiluminescence detection of adenosine triphosphate.

    Science.gov (United States)

    Wang, Ping; Zhang, Tonghuan; Yang, Taoyi; Jin, Nan; Zhao, Yanjun; Fan, Aiping

    2014-08-07

    A highly sensitive and selective chemiluminescent (CL) biosensor for adenosine triphosphate (ATP) was developed by taking advantage of the ATP-dependent enzymatic reaction (ATP-DER), the powerful signal amplification capability of rolling circle amplification (RCA), and hydroxylamine-amplified gold nanoparticles (Au NPs). The strategy relies on the ability of ATP, a cofactor of T4 DNA ligase, to trigger the ligation-RCA reaction. In the presence of ATP, the T4 DNA ligase catalyzes the ligation reaction between the two ends of the padlock probe, producing a closed circular DNA template that initiates the RCA reaction with phi29 DNA polymerase and dNTP. Therein, many complementary copies of the circular template can be generated. The ATP-DER is eventually converted into a detectable CL signal after a series of processes, including gold probe hybridization, hydroxylamine amplification, and oxidative gold metal dissolution coupled with a simple and sensitive luminol CL reaction. The CL signal is directly proportional to the ATP level. The results showed that the detection limit of the assay is 100 pM of ATP, which compares favorably with those of other ATP detection techniques. In addition, by taking advantage of ATP-DER, the proposed CL sensing system exhibits extraordinary specificity towards ATP and could distinguish the target molecule ATP from its analogues. The proposed method provides a new and versatile platform for the design of novel DNA ligation reaction-based CL sensing systems for other cofactors. This novel ATP-DER based CL sensing system may find wide applications in clinical diagnosis as well as in environmental and biomedical fields.

  17. Small sample whole-genome amplification

    Science.gov (United States)

    Hara, Christine; Nguyen, Christine; Wheeler, Elizabeth; Sorensen, Karen; Arroyo, Erin; Vrankovich, Greg; Christian, Allen

    2005-11-01

    Many challenges arise when trying to amplify and analyze human samples collected in the field due to limitations in sample quantity, and contamination of the starting material. Tests such as DNA fingerprinting and mitochondrial typing require a certain sample size and are carried out in large volume reactions; in cases where insufficient sample is present whole genome amplification (WGA) can be used. WGA allows very small quantities of DNA to be amplified in a way that enables subsequent DNA-based tests to be performed. A limiting step to WGA is sample preparation. To minimize the necessary sample size, we have developed two modifications of WGA: the first allows for an increase in amplified product from small, nanoscale, purified samples with the use of carrier DNA while the second is a single-step method for cleaning and amplifying samples all in one column. Conventional DNA cleanup involves binding the DNA to silica, washing away impurities, and then releasing the DNA for subsequent testing. We have eliminated losses associated with incomplete sample release, thereby decreasing the required amount of starting template for DNA testing. Both techniques address the limitations of sample size by providing ample copies of genomic samples. Carrier DNA, included in our WGA reactions, can be used when amplifying samples with the standard purification method, or can be used in conjunction with our single-step DNA purification technique to potentially further decrease the amount of starting sample necessary for future forensic DNA-based assays.

  18. Cascade DNA nanomachine and exponential amplification biosensing.

    Science.gov (United States)

    Xu, Jianguo; Wu, Zai-Sheng; Shen, Weiyu; Xu, Huo; Li, Hongling; Jia, Lee

    2015-11-15

    DNA is a versatile scaffold for the assembly of multifunctional nanostructures, and potential applications of various DNA nanodevices have been recently demonstrated for disease diagnosis and treatment. In the current study, a powerful cascade DNA nanomachine was developed that can execute the exponential amplification of p53 tumor suppressor gene. During the operation of the newly-proposed DNA nanomachine, dual-cyclical nucleic acid strand-displacement polymerization (dual-CNDP) was ingeniously introduced, where the target trigger is repeatedly used as the fuel molecule and the nicked fragments are dramatically accumulated. Moreover, each displaced nicked fragment is able to activate the another type of cyclical strand-displacement amplification, increasing exponentially the value of fluorescence intensity. Essentially, one target binding event can induce considerable number of subsequent reactions, and the nanodevice was called cascade DNA nanomachine. It can implement several functions, including recognition element, signaling probe, polymerization primer and template. Using the developed autonomous operation of DNA nanomachine, the p53 gene can be quantified in the wide concentration range from 0.05 to 150 nM with the detection limit of 50 pM. If taking into account the final volume of mixture, the detection limit is calculated as lower as 6.2 pM, achieving an desirable assay ability. More strikingly, the mutant gene can be easily distinguished from the wild-type one. The proof-of-concept demonstrations reported herein is expected to promote the development and application of DNA nanomachine, showing great potential value in basic biology and medical diagnosis. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Instrument for Real-Time Digital Nucleic Acid Amplification on Custom Microfluidic Devices.

    Directory of Open Access Journals (Sweden)

    David A Selck

    Full Text Available Nucleic acid amplification tests that are coupled with a digital readout enable the absolute quantification of single molecules, even at ultralow concentrations. Digital methods are robust, versatile and compatible with many amplification chemistries including isothermal amplification, making them particularly invaluable to assays that require sensitive detection, such as the quantification of viral load in occult infections or detection of sparse amounts of DNA from forensic samples. A number of microfluidic platforms are being developed for carrying out digital amplification. However, the mechanistic investigation and optimization of digital assays has been limited by the lack of real-time kinetic information about which factors affect the digital efficiency and analytical sensitivity of a reaction. Commercially available instruments that are capable of tracking digital reactions in real-time are restricted to only a small number of device types and sample-preparation strategies. Thus, most researchers who wish to develop, study, or optimize digital assays rely on the rate of the amplification reaction when performed in a bulk experiment, which is now recognized as an unreliable predictor of digital efficiency. To expand our ability to study how digital reactions proceed in real-time and enable us to optimize both the digital efficiency and analytical sensitivity of digital assays, we built a custom large-format digital real-time amplification instrument that can accommodate a wide variety of devices, amplification chemistries and sample-handling conditions. Herein, we validate this instrument, we provide detailed schematics that will enable others to build their own custom instruments, and we include a complete custom software suite to collect and analyze the data retrieved from the instrument. We believe assay optimizations enabled by this instrument will improve the current limits of nucleic acid detection and quantification, improving our

  20. Loop-mediated isothermal amplification (LAMP) assay for the diagnosis of fasciolosis in sheep and its application under field conditions

    OpenAIRE

    Mart?nez-Valladares, Mar?a; Rojo-V?zquez, Francisco Antonio

    2016-01-01

    Background Loop-mediated isothermal amplification (LAMP) is a very specific, efficient, and rapid gene amplification procedure in which the reaction can run at a constant temperature. In the current study we have developed a LAMP assay to improve the diagnosis of Fasciola spp. in the faeces of sheep. Findings After the optimisation of the LAMP assay we have shown similar results between this technique and the standard PCR using the outer primers of the LAMP reaction. In both cases the limit o...

  1. Development of a recombinase polymerase amplification assay for Vibrio parahaemolyticus detection with an internal amplification control.

    Science.gov (United States)

    Yang, Huan-Lan; Wei, Shuang; Gooneratne, Ravi; Mutukumira, Anthony N; Ma, Xue-Jun; Tang, Shu-Ze; Wu, Xi-Yang

    2018-04-01

    A novel RPA-IAC assay using recombinase polymerase and an internal amplification control (IAC) for Vibrio parahaemolyticus detection was developed. Specific primers were designed based on the coding sequence for the toxR gene in V. parahaemolyticus. The recombinase polymerase amplification (RPA) reaction was conducted at a constant low temperature of 37 °C for 20 min. Assay specificity was validated by using 63 Vibrio strains and 10 non-Vibrio bacterial species. In addition, a competitive IAC was employed to avoid false-negative results, which co-amplified simultaneously with the target sequence. The sensitivity of the assay was determined as 3 × 10 3 CFU/mL, which is decidedly more sensitive than the established PCR method. This method was then used to test seafood samples that were collected from local markets. Seven out of 53 different raw seafoods were detected as V. parahaemolyticus-positive, which were consistent with those obtained using traditional culturing method and biochemical assay. This novel RPA-IAC assay provides a rapid, specific, sensitive, and more convenient detection method for V. parahaemolyticus.

  2. Critical points and magnitude of impacts on the packing line: effect on ripening and quality of 'Packham's Triumph' pears

    Directory of Open Access Journals (Sweden)

    Josiane Pasini

    2017-06-01

    Full Text Available Pears have a very sensitive epidermis and are prone to signal mechanical blemishes, which result in reduced visual quality and low consumer acceptance. The objective of this work was to identify critical points and the magnitude of impact forces on a packing line at a commercial packinghouse. The effect of injuries on ripening and quality of ´Packham´s Triumph´ pears was also evaluated after cold storage. The packing line was scrutinized on its transfer points, fruit drop heights and cushioning overlays, which allowed to acquire the maximum accelerations on each spot. The maximum acceleration forces were reproduced in the lab with ‘Packham’s Triumph’ pears to evaluate the effects on fruit quality after cold storage. Four critical points were noticed on the packing line: at the transfer from the conveyor belt to the lifting rollers, at the transfer from the lifting rollers to the washing ramp with rotatory brushes, at the entrance to the singulator at the end of the conveyor belt and at the drop from the sizer to the packing stalls. Ripening of ‘Packham’s Triumph’ pears invariably came about during cold storage, and independently of the imposed impacts. The impacts under the circumstances of the test did not affect the quality of ´Packham´s Triumph´ pears kept for up to 120 days at cold storage followed by five days at room temperature.

  3. Evaluation of cinnamon essential oil microemulsion and its vapor phase for controlling postharvest gray mold of pears (Pyrus pyrifolia).

    Science.gov (United States)

    Wang, Yifei; Zhao, Ruipeng; Yu, Ling; Zhang, Yunbin; He, Yan; Yao, Jie

    2014-03-30

    Essential oil of cinnamon (CM) is a potential alternative to chemical fungicides. Thus this work aimed to investigate the possible effects of CM microemulsions on decay developments and qualitative properties of pears. The decay incidence of samples treated with 500 µg L⁻¹ microemulsion was significantly reduced by 18.7% in comparison to that of 500 µg L⁻¹ non-microemulsion after 4 days' storage at 20 °C. In the vapor phase, the CM microemulsion with the lowest concentration had the best control for decay incidence and lesion diameter. The interval between inoculations also influenced decay development. Pears treated with Botrytis cinerea and immediately followed by CM microemulsion showed the lowest decay incidence. Moreover, in the natural decay experiment, the percentage of rotted pears was 3.8% in the CM microemulsion treatment and 5.8% in the control. CM microemulsion delayed the loss of ascorbic acid, yet it had no significant influence on pear qualities such as firmness and color. CM microemulsion may be an alternative way to control the gray mold of pears without a negative influence on its qualities. © 2013 Society of Chemical Industry.

  4. MET amplification, expression, and exon 14 mutations in colorectal adenocarcinoma.

    Science.gov (United States)

    Zhang, Meng; Li, Guichao; Sun, Xiangjie; Ni, Shujuan; Tan, Cong; Xu, Midie; Huang, Dan; Ren, Fei; Li, Dawei; Wei, Ping; Du, Xiang

    2018-04-08

    MET amplification, expression, and splice mutations at exon 14 result in dysregulation of the MET signaling pathway. The aim of this study was to identify the relationship between MET amplification, protein or mRNA expression, and mutations in colorectal cancer (CRC). MET immunohistochemistry (IHC) was used for MET protein expression analysis and fluorescence in situ hybridization (FISH) was used for MET amplification detection. Both analyses were performed in tissue microarrays (TMA) containing 294 of colorectal adenocarcinoma tissue samples and 131 samples of adjacent normal epithelial tissue. MET mRNA expression was examined by real-time quantitative polymerase chain reaction (qRT-PCR) in 72 fresh colorectal adenocarcinoma tissue samples and adjacent normal colon tissue. PCR sequencing was performed to screen for MET exon 14 splice mutations in 59 fresh CRC tissue samples. Our results showed that MET protein expression was higher in colorectal tumor tissue than in adjacent normal intestinal epithelium. Positive MET protein expression was associated with significantly poorer overall survival (OS) and disease-free survival (DFS). Multivariate analysis revealed that positive MET protein expression was an independent risk factor for DFS, but not for OS. MET mRNA expression was upregulated in tumor tissues compared with the adjacent normal tissues. The incidence of MET amplification was 4.4%. None of the patients was positive for MET mutation. Collectively, MET was overexpressed in colorectal adenocarcinoma, and its positive protein expression predicted a poorer outcome in CRC patients. Furthermore, according to our results, MET amplification and 14 exon mutation are extremely rare events in colorectal adenocarcinoma. Copyright © 2018. Published by Elsevier Inc.

  5. Comparative transcriptomic analysis reveals that Ethylene/H2O2-mediated hypersensitive response and program cell death determine the compatible interaction of Sand pear and Alternaria Alternata

    Science.gov (United States)

    A major production restriction on sand pear (Pyrus pyrifolia) is black spot disease caused by the necrotrophic fungus Alternaria alternata. However, pear response mechanism to A. alternata is unknown at molecular level. Here, host responses of a resistant cultivar Cuiguan (CG) and a susceptible cult...

  6. Laser amplification in excited dielectrics

    DEFF Research Database (Denmark)

    Winkler, Thomas; Haahr-Lillevang, Lasse; Sarpe, Cristian

    2018-01-01

    Wide-bandgap dielectrics such as glasses or water are transparent at visible and infrared wavelengths. This changes when they are exposed to ultrashort and highly intense laser pulses. Different interaction mechanisms lead to the appearance of various transient nonlinear optical phenomena. Using...... these, the optical properties of dielectrics can be controlled from the transparent to the metal-like state. Here we expand this range by a yet unexplored mechanism in excited dielectrics: amplification. In a two-colour pump-probe experiment, we show that a 400nm femtosecond laser pulse is coherently...

  7. Resonant primordial gravitational waves amplification

    Directory of Open Access Journals (Sweden)

    Chunshan Lin

    2016-01-01

    Full Text Available We propose a mechanism to evade the Lyth bound in models of inflation. We minimally extend the conventional single-field inflation model in general relativity (GR to a theory with non-vanishing graviton mass in the very early universe. The modification primarily affects the tensor perturbation, while the scalar and vector perturbations are the same as the ones in GR with a single scalar field at least at the level of linear perturbation theory. During the reheating stage, the graviton mass oscillates coherently and leads to resonant amplification of the primordial tensor perturbation. After reheating the graviton mass vanishes and we recover GR.

  8. Whole genome amplification - Review of applications and advances

    Energy Technology Data Exchange (ETDEWEB)

    Hawkins, Trevor L.; Detter, J.C.; Richardson, Paul

    2001-11-15

    The concept of Whole Genome Amplification is something that has arisen in the past few years as modifications to the polymerase chain reaction (PCR) have been adapted to replicate regions of genomes which are of biological interest. The applications here are many--forensics, embryonic disease diagnosis, bio terrorism genome detection, ''imoralization'' of clinical samples, microbial diversity, and genotyping. The key question is if DNA can be replicated a genome at a time without bias or non random distribution of the target. Several papers published in the last year and currently in preparation may lead to the conclusion that whole genome amplification may indeed be possible and therefore open up a new avenue to molecular biology.

  9. Amplification biases: possible differences among deviating gene expressions

    Directory of Open Access Journals (Sweden)

    Piumi Francois

    2008-01-01

    Full Text Available Abstract Background Gene expression profiling has become a tool of choice to study pathological or developmental questions but in most cases the material is scarce and requires sample amplification. Two main procedures have been used: in vitro transcription (IVT and polymerase chain reaction (PCR, the former known as linear and the latter as exponential. Previous reports identified enzymatic pitfalls in PCR and IVT protocols; however the possible differences between the sequences affected by these amplification defaults were only rarely explored. Results Screening a bovine cDNA array dedicated to embryonic stages with embryonic (n = 3 and somatic tissues (n = 2, we proceeded to moderate amplifications starting from 1 μg of total RNA (global PCR or IVT one round. Whatever the tissue, 16% of the probes were involved in deviating gene expressions due to amplification defaults. These distortions were likely due to the molecular features of the affected sequences (position within a gene, GC content, hairpin number but also to the relative abundance of these transcripts within the tissues. These deviating genes mainly encoded housekeeping genes from physiological or cellular processes (70% and constituted 2 subsets which did not overlap (molecular features, signal intensities, gene ID. However, the differential expressions identified between embryonic stages were both reliable (minor intersect with biased expressions and relevant (biologically validated. In addition, the relative expression levels of those genes were biologically similar between amplified and unamplified samples. Conclusion Conversely to the most recent reports which challenged the use of intense amplification procedures on minute amounts of RNA, we chose moderate PCR and IVT amplifications for our gene profiling study. Conclusively, it appeared that systematic biases arose even with moderate amplification procedures, independently of (i the sample used: brain, ovary or embryos, (ii

  10. Compatibilidade de enxertia de cultivares de marmeleiros com pereiras Compatibility of pear cultivars on quinces rootstocks

    Directory of Open Access Journals (Sweden)

    Zeni Fonseca Pinto Tomaz

    2009-12-01

    Full Text Available A insuficiência de estudos sobre compatibilidade de porta-enxertos é um dos fatores limitantes ao desenvolvimento da cultura da pereira (Pyrus sp. no Brasil. A utilização do marmeleiro (Cydonia oblonga como porta-enxerto para a cultura da pereira apresenta inúmeras vantagens, entre as quais a redução do vigor e a rápida entrada em produção; todavia, sua combinação com algumas cultivares copa apresenta problemas de incompatibilidade de enxertia, podendo ocasionar a ruptura do caule das plantas no pomar. Objetivou-se, neste trabalho, avaliar a compatibilidade de enxertia de algumas cultivares de marmeleiros ('Quince C' e 'Adams' com pereiras ('Packham's Triumph' e 'Kieffer'. As variáveis analisadas foram: diâmetro da secção do tronco no ponto de enxertia, 5 cm abaixo e 5 cm acima do ponto de enxertia, diferença do diâmetro entre porta-enxerto e copa, altura das plantas, volume e massa seca da copa e raízes. Além disso, efetuou-se a observação da conexão vascular no ponto de enxertia através da imersão da base das plantas (abaixo do ponto de enxertia, em solução corante de Ácido Fuccínico 0,08%. Concluiu-se que a cultivar 'Packham's Triumph'apresenta compatibilidade de enxertia com o marmeleiro cultivares 'Adams'e 'Quince C', enquanto o híbrido 'Kieffer' apresentou sintomas morfológicos de incompatibilidade de enxertia com o marmeleiro cultivares 'Quince C' e 'Adams'.The lack of studies on compatibility of pear cultivars and rootstocks is one of the limiting factors on the development of the pear crop in Brazil. The use of quinces as rootstocks for pear cultivars has several advantages, among them the reduction in vigor and earlier bearing trees, however, its combination with some scions cultivars results in problems of incompatibility , such as lost of trees of the orchard due to break of the graft union. The objective of this study was to determine the compatibility between pears cvs. Packham's Triumph and Kieffer

  11. Comparison of the transcriptomic analysis between two Chinese white pear (Pyrus bretschneideri Rehd. genotypes of different stone cells contents.

    Directory of Open Access Journals (Sweden)

    Jinyun Zhang

    Full Text Available Stone cell content is thought to be one of the key determinants for fruit quality in pears. However, the molecular mechanism of stone cell development remains poorly understood. In this study, we found that the stone cell clusters (SCCs distribution and area in 'Dangshan Su' (with abundant stone cells were higher as compared to 'Lianglizaosu' (low stone cell content bud sport of 'Dangshan Su' based on the histochemical staining, and the correlations of lignin content with stone cell content and SCC area was significant. The fruits of 'Dangshan Su' and 'Lianglizaosu' at three different developmental stages (23 and 55 days after flowering and mature were sampled for comparative transcriptome analysis to explore the metabolic pathways associated with stone cell development. A total of 42444 unigenes were obtained from two varieties, among which 7203 differentially expressed genes (DEGs were identified by comparison of the six transcriptomes. Specifically, many DEGs associated with lignin biosynthesis were identified, including coumaroylquinate 3-monooxygenase (C3H, shikimate O-hydroxycinnamoyltransferase (HCT, ferulate 5-hydroxylase (F5H, cinnamyl alcohol dehydrogenase (CAD and peroxidase (POD, as well as genes related to carbon metabolism, such as sorbitol dehydrogenase-like (SDH-like and ATP-dependent 6-phosphofructokinase (ATP-PFK. At the peak of the stone cell content (55 days after flowering, the expression level of these genes in 'Dangshan Su' was significantly increased compared with 'Lianglizaosu', indicating that these genes were closely related to stone cell development. We validated the transcriptional levels of 33 DEGs using quantitative real-time polymerase chain reaction (qRT-PCR analysis. The results were consistent with the transcriptome analysis, indicating the reliability of transcriptome data. In addition, subcellular localization analysis of three DEGs in lignin synthesis (PbC3H, PbF5H and PbPOD revealed that these proteins are

  12. Balsam-Pear-Skin-Like Porous Polyacrylonitrile Nanofibrous Membranes Grafted with Polyethyleneimine for Postcombustion CO2 Capture.

    Science.gov (United States)

    Zhang, Yufei; Guan, Jiming; Wang, Xianfeng; Yu, Jianyong; Ding, Bin

    2017-11-22

    Amine-containing sorbents have been extensively studied for postcombustion carbon dioxide (CO 2 ) capture because of their ability to chemisorb CO 2 from the flue gas. However, most sorbents are in the form of powders currently, which is not the ideal configuration for the flue gas separation because of the fragile nature and poor mechanical properties, resulting in blocking of the flow pipes and difficult recycling. Herein, we present a novel approach for the facile fabrication of flexible, robust, and polyethyleneimine-grafted (PEI-grafted) hydrolyzed porous PAN nanofibrous membranes (HPPAN-PEI NFMs) through the combination of electrospinning, pore-forming process, hydrolysis reaction, and the subsequent grafting technique. Excitingly, we find that all the resultant porous PAN (PPAN) fibers exhibit a balsam-pear-skin-like porous structure due to the selective removal of poly(vinylpyrrolidone) (PVP) from PAN/PVP fibers by water extraction. Significantly, the HPPAN-PEI NFMs retain their mesoporosity, as well as exhibit good thermal stability and prominent tensile strength (11.1 MPa) after grafting, guaranteeing their application in CO 2 trapping from the flue gas. When exposed to CO 2 at 40 °C, the HPPAN-PEI NFMs show an enhanced CO 2 adsorption capacity of 1.23 mmol g -1 (based on the overall quantity of the sample) or 6.15 mmol g -1 (based on the quantity of grafted PEI). Moreover, the developed HPPAN-PEI NFMs display significantly selective capture for CO 2 over N 2 and excellent recyclability. The CO 2 capacity retains 92% of the initial value after 20 adsorption-desorption cycle tests, indicating that the resultant HPPAN-PEI NFMs have good long-term stability. This work paves the way for fabricating NFM-based solid adsorption materials endowed with a porous structure applied to efficient postcombustion CO 2 capture.

  13. Single primer amplification reaction methods reveal exotic and ...

    Indian Academy of Sciences (India)

    Unknown

    Despite economic importance, the origin and domestication of mulberry has not ..... The numbers at the nodes in each tree are the bootstrap percent values (only values greater than or equal to 50 are shown) for the ..... Dandin S B 1998 Mulberry: A versatile biosource in the service of mankind; Acta Sericologica Sinica 24 ...

  14. Risk Perception and Social Amplification

    International Nuclear Information System (INIS)

    Smith, R.E.

    2001-01-01

    This paper seeks to consider social amplification as it applies to risk perception. Perceptions of the magnitude of a risk are conditioned by issues such as the degree of uncertainty in probability and consequences, the nature of the consequences and the relative weightings placed on probability and consequences. Risk perceptions are also influenced by factors such as confidence in the operator of an industrial process, trust in the regulator and the perceived fairness of regulatory decision-making. Different people may hold different views about these issues and there may also be difficulties in communication. The paper identifies and discusses self-reinforcing mechanisms, which will be labelled 'lock-in' here. They appear to apply in many situations where social amplification is observed. Historically, the term 'lock-in' has been applied mainly in the technological context but, in this paper, four types of lock-in are identified, namely scientific/technological, economic, social and institutional lock-in. One type of lock-in tends to lead to the next and all are buttressed by people's general acceptance of the familiar, fear of the unknown and resistance to change. The regulator seeks to make decisions which achieve the common good rather than supporting or perpetuating any set of vested interests. In this regard the locked-in positions of stakeholders, whether organisations, interest groups, or individual members of the public, are obstacles and challenges. Existing methods of consultation are unsatisfactory in terms of achieving a proper and productive level of dialogue with stakeholders

  15. MYB Transcription Factors in Chinese Pear (Pyrus bretschneideri Rehd.: Genome-Wide Identification, Classification and Expression Profiling during Fruit Development

    Directory of Open Access Journals (Sweden)

    Yun Peng eCao

    2016-04-01

    Full Text Available The MYB family is one of the largest families of transcription factors in plants. Although some MYBs have been reported to play roles in secondary metabolism, no comprehensive study of the MYB family in Chinese pear (Pyrus bretschneideri Rehd. has been reported. In the present study, we performed genome-wide analysis of MYB genes in Chinese pear, designated as PbMYBs, including analyses of their phylogenic relationships, structures, chromosomal locations, promoter regions, GO annotations and collinearity. A total of 129 PbMYB genes were identified in the pear genome and were divided into 31 subgroups based on phylogenetic analysis. These PbMYBs were unevenly distributed among 16 chromosomes (total of 17 chromosomes. The occurrence of gene duplication events indicated that whole-genome duplication and segmental duplication likely played key roles in expansion of the PbMYB gene family. Ka/Ks analysis suggested that the duplicated PbMYBs mainly experienced purifying selection with restrictive functional divergence after the duplication events. Interspecies microsynteny analysis revealed maximum orthology between pear and peach, followed by plum and strawberry. Subsequently, the expression patterns of 20 PbMYB genes that may be involved in lignin biosynthesis according to their phylogenetic relationships were examined throughout fruit development. Among the twenty genes examined, PbMYB25 and PbMYB52 exhibited expression patterns consistent with the typical variations in the lignin content previously reported. Moreover, sub-cellular localization analysis revealed that two proteins PbMYB25 and PbMYB52 were localized to the nucleus. All together, PbMYB25 and PbMYB52 were inferred to be candidate genes involved in the regulation of lignin biosynthesis during the development of pear fruit. This study provides useful information for further functional analysis of the MYB gene family in pear.

  16. Hurdle technology applied to prickly pear beverages for inhibiting Saccharomyces cerevisiae and Escherichia coli.

    Science.gov (United States)

    García-García, R; Escobedo-Avellaneda, Z; Tejada-Ortigoza, V; Martín-Belloso, O; Valdez-Fragoso, A; Welti-Chanes, J

    2015-06-01

    The effect of pH reduction (from 6·30-6·45 to 4·22-4·46) and the addition of antimicrobial compounds (sodium benzoate and potassium sorbate) on the inhibition of Saccharomyces cerevisiae and Escherichia coli in prickly pear beverages formulated with the pulp and peel of Villanueva (V, Opuntia albicarpa) and Rojo Vigor (RV, Opuntia ficus-indica) varieties during 14 days of storage at 25°C, was evaluated. RV variety presented the highest microbial inhibition. By combining pH reduction and preservatives, reductions of 6·2-log10 and 2·3-log10 for E. coli and S. cerevisiae were achieved respectively. Due to the low reduction of S. cerevisiae, pulsed electric fields (PEF) (11-15 μs/25-50 Hz/27-36 kV cm(-1)) was applied as another preservation factor. The combination of preservatives, pH reduction and PEF at 13-15 μs/25-50 Hz for V variety, and 11 μs/50 Hz, 13-15 μs/25-50 Hz for RV, had a synergistic effect on S. cerevisiae inhibition, achieving at least 3·4-log10 of microbial reduction immediately after processing, and more than 5-log10 at fourth day of storage at 25°C maintained this reduction during 21 days of storage (P > 0·05). Hurdle technology using PEF in combination with other factors is adequate to maintain stable prickly pear beverages during 21 days/25°C. Significance and impact of the study: Prickly pear is a fruit with functional value, with high content of nutraceuticals and antioxidant activity. Functional beverages formulated with the pulp and peel of this fruit represent an alternative for its consumption. Escherichia coli and Saccharomyces cerevisiae are micro-organisms that typically affect fruit beverage quality and safety. The food industry is looking for processing technologies that maintain quality without compromising safety. Hurdle technology, including pulsed electric fields (PEF) could be an option to achieve this. The combination of PEF, pH reduction and preservatives is an alternative to obtain safe and minimally processed

  17. Clarification of purple cactus pear juice using microfiltration membranes to obtain a solution of betalain pigments

    Directory of Open Access Journals (Sweden)

    Cristina VERGARA

    2015-09-01

    Full Text Available Summary Betalains are fruit pigments possessing health-giving properties. To isolate the pigments, the juice must be separated from the fruit matrix, which contains biopolymers. The aim of this study was to clarify cactus pear juice by microfiltration to obtain a clarified juice containing betalains. For this purpose, two 0.2 µm pore size microfiltration membranes (ceramic and polymeric were tested. The permeates were clear, free of turbidity and high in betalains (20%, also containing polyphenols and antioxidant activity, whereas the retained fractions were high in mucilage. The best separation was obtained using the ceramic membrane.

  18. Genetic, metabolite and developmental determinism of fruit friction discolouration in pear.

    Science.gov (United States)

    Saeed, Munazza; Brewer, Lester; Johnston, Jason; McGhie, Tony K; Gardiner, Susan E; Heyes, Julian A; Chagné, David

    2014-09-16

    The unattractive appearance of the surface of pear fruit caused by the postharvest disorder friction discolouration (FD) is responsible for significant consumer dissatisfaction in markets, leading to lower returns to growers. Developing an understanding of the genetic control of FD is essential to enable the full application of genomics-informed breeding for the development of new pear cultivars. Biochemical constituents [phenolic compounds and ascorbic acid (AsA)], polyphenol oxidase (PPO) activity, as well as skin anatomy, have been proposed to play important roles in FD susceptibility in studies on a limited number of cultivars. However, to date there has been no investigation on the biochemical and genetic control of FD, employing segregating populations. In this study, we used 250 seedlings from two segregating populations (POP369 and POP356) derived from interspecific crosses between Asian (Pyrus pyrifolia Nakai and P. bretschneideri Rehd.) and European (P. communis) pears to identify genetic factors associated with susceptibility to FD. Single nucleotide polymorphism (SNP)-based linkage maps suitable for QTL analysis were developed for the parents of both populations. The maps for population POP369 comprised 174 and 265 SNP markers for the male and female parent, respectively, while POP356 maps comprised 353 and 398 SNP markers for the male and female parent, respectively. Phenotypic data for 22 variables were measured over two successive years (2011 and 2012) for POP369 and one year (2011) only for POP356. A total of 221 QTLs were identified that were linked to 22 phenotyped variables, including QTLs associated with FD for both populations that were stable over the successive years. In addition, clear evidence of the influence of developmental factors (fruit maturity) on FD and other variables was also recorded. The QTLs associated with fruit firmness, PPO activity, AsA concentration and concentration of polyphenol compounds as well as FD are the first

  19. Allergenic pollens and spores in the working environment of Japanese pear farmers.

    Science.gov (United States)

    Teranishi, H; Uchida, M; Hayashi, S; Yamada, N

    2007-01-01

    Occupational allergies such as pollinosis are reported in several agricultural works in Japan. Many pollens and spores were observed in Japanese pear orchard during the artificial pollination season. By the study on daily symptoms in an allergic farmer, we confirmed that the pollinosis symptoms were most common and most severe during the artificial pollination. These results suggest that the exposure to allergenic pollens and spores induces allergic symptoms. Thus, caution should be paid for the avoidance of the exposure to these allergenic pollens and spores to prevent the allergy.

  20. Antioxidant betalains from cactus pear (Opuntia ficus-indica) inhibit endothelial ICAM-1 expression.

    Science.gov (United States)

    Gentile, C; Tesoriere, L; Allegra, M; Livrea, M A; D'Alessio, P

    2004-12-01

    It has been suggested that some pigments would have antioxidant properties and that their presence in dietary constituents would contribute to reduce the risk of oxidative stress-correlated diseases. Among others, inflammatory response depends on redox status and may implicate oxidative stress. Vascular endothelial cells are a direct target of oxidative stress in inflammation. We have tested the impact of the free radical scavenger and antioxidant properties of betalains from the prickle pear in an in vitro model of endothelial cells. Here we show the capacity of betalains to protect endothelium from cytokine-induced redox state alteration, through ICAM-1 inhibition.

  1. Strand Invasion Based Amplification (SIBA®): a novel isothermal DNA amplification technology demonstrating high specificity and sensitivity for a single molecule of target analyte.

    Science.gov (United States)

    Hoser, Mark J; Mansukoski, Hannu K; Morrical, Scott W; Eboigbodin, Kevin E

    2014-01-01

    Isothermal nucleic acid amplification technologies offer significant advantages over polymerase chain reaction (PCR) in that they do not require thermal cycling or sophisticated laboratory equipment. However, non-target-dependent amplification has limited the sensitivity of isothermal technologies and complex probes are usually required to distinguish between non-specific and target-dependent amplification. Here, we report a novel isothermal nucleic acid amplification technology, Strand Invasion Based Amplification (SIBA). SIBA technology is resistant to non-specific amplification, is able to detect a single molecule of target analyte, and does not require target-specific probes. The technology relies on the recombinase-dependent insertion of an invasion oligonucleotide (IO) into the double-stranded target nucleic acid. The duplex regions peripheral to the IO insertion site dissociate, thereby enabling target-specific primers to bind. A polymerase then extends the primers onto the target nucleic acid leading to exponential amplification of the target. The primers are not substrates for the recombinase and are, therefore unable to extend the target template in the absence of the IO. The inclusion of 2'-O-methyl RNA to the IO ensures that it is not extendible and that it does not take part in the extension of the target template. These characteristics ensure that the technology is resistant to non-specific amplification since primer dimers or mis-priming are unable to exponentially amplify. Consequently, SIBA is highly specific and able to distinguish closely-related species with single molecule sensitivity in the absence of complex probes or sophisticated laboratory equipment. Here, we describe this technology in detail and demonstrate its use for the detection of Salmonella.

  2. Strand Invasion Based Amplification (SIBA®: a novel isothermal DNA amplification technology demonstrating high specificity and sensitivity for a single molecule of target analyte.

    Directory of Open Access Journals (Sweden)

    Mark J Hoser

    Full Text Available Isothermal nucleic acid amplification technologies offer significant advantages over polymerase chain reaction (PCR in that they do not require thermal cycling or sophisticated laboratory equipment. However, non-target-dependent amplification has limited the sensitivity of isothermal technologies and complex probes are usually required to distinguish between non-specific and target-dependent amplification. Here, we report a novel isothermal nucleic acid amplification technology, Strand Invasion Based Amplification (SIBA. SIBA technology is resistant to non-specific amplification, is able to detect a single molecule of target analyte, and does not require target-specific probes. The technology relies on the recombinase-dependent insertion of an invasion oligonucleotide (IO into the double-stranded target nucleic acid. The duplex regions peripheral to the IO insertion site dissociate, thereby enabling target-specific primers to bind. A polymerase then extends the primers onto the target nucleic acid leading to exponential amplification of the target. The primers are not substrates for the recombinase and are, therefore unable to extend the target template in the absence of the IO. The inclusion of 2'-O-methyl RNA to the IO ensures that it is not extendible and that it does not take part in the extension of the target template. These characteristics ensure that the technology is resistant to non-specific amplification since primer dimers or mis-priming are unable to exponentially amplify. Consequently, SIBA is highly specific and able to distinguish closely-related species with single molecule sensitivity in the absence of complex probes or sophisticated laboratory equipment. Here, we describe this technology in detail and demonstrate its use for the detection of Salmonella.

  3. Tiny Grains Give Huge Gains: Nanocrystal–Based Signal Amplification for Biomolecule Detection

    Science.gov (United States)

    Tong, Sheng; Ren, Binbin; Zheng, Zhilan; Shen, Han; Bao, Gang

    2013-01-01

    Nanocrystals, despite their tiny sizes, contain thousands to millions of atoms. Here we show that the large number of atoms packed in each metallic nanocrystal can provide a huge gain in signal amplification for biomolecule detection. We have devised a highly sensitive, linear amplification scheme by integrating the dissolution of bound nanocrystals and metal-induced stoichiometric chromogenesis, and demonstrated that signal amplification is fully defined by the size and atom density of nanocrystals, which can be optimized through well-controlled nanocrystal synthesis. Further, the rich library of chromogenic reactions allows implementation of this scheme in various assay formats, as demonstrated by the iron oxide nanoparticle linked immunosorbent assay (ILISA) and blotting assay developed in this study. Our results indicate that, owing to the inherent simplicity, high sensitivity and repeatability, the nanocrystal based amplification scheme can significantly improve biomolecule quantification in both laboratory research and clinical diagnostics. This novel method adds a new dimension to current nanoparticle-based bioassays. PMID:23659350

  4. Combined Analyses of Chloroplast DNA Haplotypes and Microsatellite Markers Reveal New Insights Into the Origin and Dissemination Route of Cultivated Pears Native to East Asia

    Directory of Open Access Journals (Sweden)

    Xiaoyan Yue

    2018-05-01

    Full Text Available Asian pear plays an important role in the world pear industry, accounting for over 70% of world total production volume. Commercial Asian pear production relies on four major pear cultivar groups, Japanese pear (JP, Chinese white pear (CWP, Chinese sand pear (CSP, and Ussurian pear (UP, but their origins remain controversial. We estimated the genetic diversity levels and structures in a large sample of existing local cultivars to investigate the origins of Asian pears using twenty-five genome-covering nuclear microsatellite (simple sequence repeats, nSSR markers and two non-coding chloroplast DNA (cpDNA regions (trnL-trnF and accD-psaI. High levels of genetic diversity were detected for both nSSRs (HE = 0.744 and cpDNAs (Hd = 0.792. The major variation was found within geographic populations of cultivated pear groups, demonstrating a close relationship among cultivar groups. CSPs showed a greater genetic diversity than CWPs and JPs, and lowest levels of genetic differentiation were detected among them. Phylogeographical analyses indicated that the CSP, CWP, and JP were derived from the same progenitor of Pyrus pyrifolia in China. A dissemination route of cultivated P. pyrifolia estimated by approximate Bayesian computation suggested that cultivated P. pyrifolia from the Middle Yangtze River Valley area contributed the major genetic resources to the cultivars, excluding those of southwestern China. Three major genetic groups of cultivated Pyrus pyrifolia were revealed using nSSRs and a Bayesian statistical inference: (a JPs; (b cultivars from South-Central China northward to northeastern China, covering the main pear production area in China; (c cultivars from southwestern China to southeastern China, including Yunnan, Guizhou, Guangdong, Guangxi, and Fujian Provinces. This reflected the synergistic effects of ecogeographical factors and human selection during cultivar spread and improvement. The analyses indicated that UP cultivars might be

  5. Multiscale image contrast amplification (MUSICA)

    Science.gov (United States)

    Vuylsteke, Pieter; Schoeters, Emile P.

    1994-05-01

    This article presents a novel approach to the problem of detail contrast enhancement, based on multiresolution representation of the original image. The image is decomposed into a weighted sum of smooth, localized, 2D basis functions at multiple scales. Each transform coefficient represents the amount of local detail at some specific scale and at a specific position in the image. Detail contrast is enhanced by non-linear amplification of the transform coefficients. An inverse transform is then applied to the modified coefficients. This yields a uniformly contrast- enhanced image without artefacts. The MUSICA-algorithm is being applied routinely to computed radiography images of chest, skull, spine, shoulder, pelvis, extremities, and abdomen examinations, with excellent acceptance. It is useful for a wide range of applications in the medical, graphical, and industrial area.

  6. Measuring the amplification of attention.

    Science.gov (United States)

    Blaser, E; Sperling, G; Lu, Z L

    1999-09-28

    An ambiguous motion paradigm, in which the direction of apparent motion is determined by salience (i.e., the extent to which an area is perceived as figure versus ground), is used to assay the amplification of color by attention to color. In the red-green colored gratings used in these experiments, without attention instructions, salience depends on the chromaticity difference between colored stripes embedded in the motion sequence and the yellow background. Selective attention to red (or to green) alters the perceived direction of motion and is found to be equivalent to increasing the physical redness (or greenness) by 25-117%, depending on the observer and color. Whereas attention to a color drastically alters the salience of that color, it leaves color appearance unchanged. A computational model, which embodies separate, parallel pathways for object perception and for salience, accounts for 99% of the variance of the experimental data.

  7. Construction and optimization of an efficient breathing-based isothermal emulsion amplification method

    Energy Technology Data Exchange (ETDEWEB)

    Shen, Yanting, E-mail: shenyanting798@126.com [Research Center for Learning Science, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); State Key Laboratory of Bioelectronics, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Tian, Fei, E-mail: 642807827@qq.com [Research Center for Learning Science, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Tu, Jing, E-mail: jtu@seu.edu.cn [State Key Laboratory of Bioelectronics, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Li, Rui, E-mail: lirui901113@163.com [Research Center for Learning Science, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Chen, Zhenzhu, E-mail: zzchen_seu@163.com [Research Center for Learning Science, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Bai, Yunfei, E-mail: whitecf@seu.edu.cn [State Key Laboratory of Bioelectronics, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Ge, Qinyu, E-mail: geqinyu@seu.edu.cn [State Key Laboratory of Bioelectronics, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Lu, Zuhong, E-mail: zhlu@seu.edu.cn [State Key Laboratory of Bioelectronics, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China)

    2017-06-22

    The reaction temperature is one of the main factors that affect the stability of emulsion PCR (emPCR). Focusing on this point, we applied the “DNA breathing” mechanism in BEAMing (Bead, Emulsion, Amplification, and Magnetic) and proposed a more stable emulsion amplification method. Compared to the conventional emPCR, this method provided excellent results. Firstly, more stable emulsion system resulted in higher percentage of single-molecular amplifications (73.17%). Secondly, an ordinary temperature-controlling device was enough. Our outcome showed that the reaction temperature of this method was not strict so that the ordinary temperature-controlling device was enough for it (the heat block sets vs. the PCR instrument: 13.140 ± 0.110 vs. 13.008 ± 0.039, P = 0.120). Thirdly, the single-biotinylated emP{sub 1} coated streptavidin beads were stable enough to be used for this method (the control temperature vs. the reaction temperature: 2967.91 ± 409.045 vs. 3026.22 ± 442.129, P = 0.334), which could replace the double-biotinylated emP{sub 1} coated beads and was benefit for saving cost. In conclusion, the method presented here with stable emulsion system, simplified temperature-controlling device, and decreased investment would be a highly streamlined and inexpensive option for future single-molecular amplification based researches. - Highlights: • A breathing-based isothermal emulsion amplification (BIEA) method was developed. • BIEA showed excellent properties compared with conventional amplification method. • Terminal breathing of DNA duplex was firstly used in emulsion amplification.

  8. Risk Perception and Social Amplification

    Energy Technology Data Exchange (ETDEWEB)

    Smith, R.E. [Environment Agency (United Kingdom)

    2001-07-01

    This paper seeks to consider social amplification as it applies to risk perception. Perceptions of the magnitude of a risk are conditioned by issues such as the degree of uncertainty in probability and consequences, the nature of the consequences and the relative weightings placed on probability and consequences. Risk perceptions are also influenced by factors such as confidence in the operator of an industrial process, trust in the regulator and the perceived fairness of regulatory decision-making. Different people may hold different views about these issues and there may also be difficulties in communication. The paper identifies and discusses self-reinforcing mechanisms, which will be labelled 'lock-in' here. They appear to apply in many situations where social amplification is observed. Historically, the term 'lock-in' has been applied mainly in the technological context but, in this paper, four types of lock-in are identified, namely scientific/technological, economic, social and institutional lock-in. One type of lock-in tends to lead to the next and all are buttressed by people's general acceptance of the familiar, fear of the unknown and resistance to change. The regulator seeks to make decisions which achieve the common good rather than supporting or perpetuating any set of vested interests. In this regard the locked-in positions of stakeholders, whether organisations, interest groups, or individual members of the public, are obstacles and challenges. Existing methods of consultation are unsatisfactory in terms of achieving a proper and productive level of dialogue with stakeholders.

  9. Cultivar and Harvest Month Influence the Nutrient Content of Opuntia spp. Cactus Pear Cladode Mucilage Extracts

    Directory of Open Access Journals (Sweden)

    Alba du Toit

    2018-04-01

    Full Text Available Mucilage extracted from cactus pear cladodes is a hydrocolloid gum. It is a novel, natural, low-kilojoule, cost-effective texture-modifying ingredient in functional food products. Yet, the cultivar with the most optimal nutrient content and the preferred harvest times are as yet unknown. For this reason, mucilage from three Opuntia ficus-indica (Algerian, Morado and Gymno-Carpo and one Opuntia robusta (Robusta cultivar were investigated to determine their nutrient content over six months. Nutrients that contribute energy (10.2 kJ/g were low. The mineral content was high (ash 17.7/100 g, particularly calcium (3.0 g/100 g and phosphorous (109.5 mg/kg. Low insoluble acid-detergent fibre (1.4 g/kg and neutral-detergent fibre (2.1 g/kg values indicated that mucilage was mostly soluble fibre. Calcium oxalate crystals were not detected in dried mucilage. Opuntia robusta powders had higher protein, extractable fat and potassium content, while Opuntia ficus-indica mucilage powders had higher polyunsaturated (Linoleic and α-Linolenic acid fat content. O. robusta Robusta mucilage, harvested after the fruit harvest (February had the lowest energy content and the highest mineral and protein content. Mucilage powders were highly soluble, low-kilojoule and mineral-rich. This is a functional ingredient that is produced from an easily cultivated crop, as cactus pears grow in areas with poor soil, extremely high daytime temperatures and limited water supplies.

  10. Population structure of Aphis spiraecola (Hemiptera: Aphididae) on pear trees in China identified using microsatellites.

    Science.gov (United States)

    Cao, Jinjun; Li, Jie; Niu, Jianqun; Liu, Xiaoxia; Zhang, Qingwen

    2012-04-01

    The spiraea aphid (Aphis spiraecola Patch) is a primary pest of fruit trees, particularly pear trees in China. Despite the economic importance of this pest, little is known about its genetic structure or its patterns of dispersal at local and regional scales; however, knowledge of these characteristics is important for establishing effective control strategies for this pest. The genetic variability of 431 individuals from 21 populations on pear trees in China was investigated using eight polymorphic microsatellite loci. The high polymorphism of these markers was evident from the expected heterozygosity value (He = 0.824) and the Polymorphism Information Content (PIC = 0.805), indicating that the spiraea aphid maintains a high level of genetic diversity. The analysis of molecular variance revealed a middle level of population differentiation (F(ST) = 0.1478) among A. spiraecola populations. This result is consistent with the results of the STRUCTURE analysis (K = 3), the unweighted pair-group method with arithmetic average tree and the Mantel test (r = 0.6392; P < 0.05). Our results indicate high levels of genetic exchange in the spiraea aphid, possibly facilitated by geography and climate. Our findings emphasize the importance of considering regional differences in studies of population structure, even when strong isolation-by-distance influences the genetic population structure of species.

  11. Cultivar and Harvest Month Influence the Nutrient Content of Opuntia spp. Cactus Pear Cladode Mucilage Extracts.

    Science.gov (United States)

    du Toit, Alba; de Wit, Maryna; Hugo, Arno

    2018-04-16

    Mucilage extracted from cactus pear cladodes is a hydrocolloid gum. It is a novel, natural, low-kilojoule, cost-effective texture-modifying ingredient in functional food products. Yet, the cultivar with the most optimal nutrient content and the preferred harvest times are as yet unknown. For this reason, mucilage from three Opuntia ficus-indica (Algerian, Morado and Gymno-Carpo) and one Opuntia robusta (Robusta) cultivar were investigated to determine their nutrient content over six months. Nutrients that contribute energy (10.2 kJ/g) were low. The mineral content was high (ash 17.7/100 g), particularly calcium (3.0 g/100 g) and phosphorous (109.5 mg/kg). Low insoluble acid-detergent fibre (1.4 g/kg) and neutral-detergent fibre (2.1 g/kg) values indicated that mucilage was mostly soluble fibre. Calcium oxalate crystals were not detected in dried mucilage. Opuntia robusta powders had higher protein, extractable fat and potassium content, while Opuntia ficus-indica mucilage powders had higher polyunsaturated (Linoleic and α-Linolenic acid) fat content. O. robusta Robusta mucilage, harvested after the fruit harvest (February) had the lowest energy content and the highest mineral and protein content. Mucilage powders were highly soluble, low-kilojoule and mineral-rich. This is a functional ingredient that is produced from an easily cultivated crop, as cactus pears grow in areas with poor soil, extremely high daytime temperatures and limited water supplies.

  12. VEGETATIVE DEVELOPMENT OF EUROPEAN PEAR WITH QUINCE AND DIFFERENT APPLICATION FORMS OF NUTRIENTS

    Directory of Open Access Journals (Sweden)

    FABIANE NUNES SILVEIRA

    Full Text Available ABSTRACT The pear consumption in Brazil is significant and the country is dependent on the import of this fruit to meet the internal market. The aim of this study was to evaluate the vegetative growth of European pear trees grafted on quince with two application forms of nutrients. The study was conducted during the 2012/13, 2013/14 and 2014/15 harvests, in the experimental area of the Universidade do Estado de Santa Catarina / UDESC, in Lages. The cultivars used were Rocha, Santa Maria and Abbé Fétel. The quinces were ‘Adams’ and ‘EMA’. The application forms of the nutrients were conventional and fertigation. The experiment was carried out in a randomized block design and the arrangement of treatments was sub subdivided plots. The application form of nutrients and rootstocks of quince little influenced on the vegetative growth of the plants, being more influenced by the scion in the soil conditions of this study.

  13. Diagnosis of brugian filariasis by loop-mediated isothermal amplification.

    Directory of Open Access Journals (Sweden)

    Catherine B Poole

    Full Text Available In this study we developed and evaluated a Brugia Hha I repeat loop-mediated isothermal amplification (LAMP assay for the rapid detection of Brugia genomic DNA. Amplification was detected using turbidity or fluorescence as readouts. Reactions generated a turbidity threshold value or a clear visual positive within 30 minutes using purified genomic DNA equivalent to one microfilaria. Similar results were obtained using DNA isolated from blood samples containing B. malayi microfilariae. Amplification was specific to B. malayi and B. timori, as no turbidity was observed using DNA from the related filarial parasites Wuchereria bancrofti, Onchocerca volvulus or Dirofilaria immitis, or from human or mosquito. Furthermore, the assay was most robust using a new strand-displacing DNA polymerase termed Bst 2.0 compared to wild-type Bst DNA polymerase, large fragment. The results indicate that the Brugia Hha I repeat LAMP assay is rapid, sensitive and Brugia-specific with the potential to be developed further as a field tool for diagnosis and mapping of brugian filariasis.

  14. Improved multiple displacement amplification (iMDA) and ultraclean reagents.

    Science.gov (United States)

    Motley, S Timothy; Picuri, John M; Crowder, Chris D; Minich, Jeremiah J; Hofstadler, Steven A; Eshoo, Mark W

    2014-06-06

    Next-generation sequencing sample preparation requires nanogram to microgram quantities of DNA; however, many relevant samples are comprised of only a few cells. Genomic analysis of these samples requires a whole genome amplification method that is unbiased and free of exogenous DNA contamination. To address these challenges we have developed protocols for the production of DNA-free consumables including reagents and have improved upon multiple displacement amplification (iMDA). A specialized ethylene oxide treatment was developed that renders free DNA and DNA present within Gram positive bacterial cells undetectable by qPCR. To reduce DNA contamination in amplification reagents, a combination of ion exchange chromatography, filtration, and lot testing protocols were developed. Our multiple displacement amplification protocol employs a second strand-displacing DNA polymerase, improved buffers, improved reaction conditions and DNA free reagents. The iMDA protocol, when used in combination with DNA-free laboratory consumables and reagents, significantly improved efficiency and accuracy of amplification and sequencing of specimens with moderate to low levels of DNA. The sensitivity and specificity of sequencing of amplified DNA prepared using iMDA was compared to that of DNA obtained with two commercial whole genome amplification kits using 10 fg (~1-2 bacterial cells worth) of bacterial genomic DNA as a template. Analysis showed >99% of the iMDA reads mapped to the template organism whereas only 0.02% of the reads from the commercial kits mapped to the template. To assess the ability of iMDA to achieve balanced genomic coverage, a non-stochastic amount of bacterial genomic DNA (1 pg) was amplified and sequenced, and data obtained were compared to sequencing data obtained directly from genomic DNA. The iMDA DNA and genomic DNA sequencing had comparable coverage 99.98% of the reference genome at ≥1X coverage and 99.9% at ≥5X coverage while maintaining both balance

  15. Systematic Analysis of the 4-Coumarate:Coenzyme A Ligase (4CL Related Genes and Expression Profiling during Fruit Development in the Chinese Pear

    Directory of Open Access Journals (Sweden)

    Yunpeng Cao

    2016-10-01

    Full Text Available In plants, 4-coumarate:coenzyme A ligases (4CLs, comprising some of the adenylate-forming enzymes, are key enzymes involved in regulating lignin metabolism and the biosynthesis of flavonoids and other secondary metabolites. Although several 4CL-related proteins were shown to play roles in secondary metabolism, no comprehensive study on 4CL-related genes in the pear and other Rosaceae species has been reported. In this study, we identified 4CL-related genes in the apple, peach, yangmei, and pear genomes using DNATOOLS software and inferred their evolutionary relationships using phylogenetic analysis, collinearity analysis, conserved motif analysis, and structure analysis. A total of 149 4CL-related genes in four Rosaceous species (pear, apple, peach, and yangmei were identified, with 30 members in the pear. We explored the functions of several 4CL and acyl-coenzyme A synthetase (ACS genes during the development of pear fruit by quantitative real-time PCR (qRT-PCR. We found that duplication events had occurred in the 30 4CL-related genes in the pear. These duplicated 4CL-related genes are distributed unevenly across all pear chromosomes except chromosomes 4, 8, 11, and 12. The results of this study provide a basis for further investigation of both the functions and evolutionary history of 4CL-related genes.

  16. Response of the pearly eye melon fly Bactrocera cucurbitae (Coquillett)(Diptera:Tephritidae) mutant to host-associated visual cues

    Science.gov (United States)

    We report on a pearly eye mutant (PEM) line generated from a single male Bactrocera cucurbitae collected in Kapoho, Hawaii. Crossing experiments with colony wild-type flies indicate that the locus controlling this trait is autosomal and the mutant allele is recessive. Experiments with females to ass...

  17. Systematics of the east Palaearctic pear psyllids (Hemiptera: Psylloidea) with particular focus on the Japanese and Korean fauna.

    Science.gov (United States)

    Cho, Geonho; Burckhardt, Daniel; Inoue, Hiromitsu; Luo, Xinyu; Lee, Seunghwan

    2017-12-04

    The confused taxonomy of the east Palaearctic pear psyllids, serious pests on cultivated pear, is reviewed. Fifty-six nominal species have been reported from Pyrus, 25 of which we consider valid and ten as not being associated with Pyrus. Our taxonomic revision suggests that, in Korea, four Cacopsylla species develop on pear: the univoltine C. burckhardti Luo et al. previously misidentified as C. pyrisuga (Foerster), the polyvoltine, seasonally dimorphic C. jukyungi (Kwon) (winter form 'cinereosignata' Luo et al., summer form 'jukyungi'), commonly found in Korean pear orchards, and C. maculatili Li (winter form 'maculatili', summer form 'qiuzili' Li) previously misidentified as C. pyricola (Foerster) by some authors, as well as the probably polyvoltine but not dimorphic C. sandolbaea (Park & Lee). The former three species (C. burckhardti, C. jukyungi, misidentified as C. chinensis (Yang & Li), and C. maculatili) occur also in Japan. Keys to the adult and fifth instar immatures as well as short biological notes are provided, and C. jukyungi and C. sandolbaea are redescribed. Following nomenclatorial changes are proposed: Cacopsylla betulaefoliae (Yang & Li, 1981) = Psylla heterobetulaefoliae Yang & Li, 1981, syn. nov.; Cacopsylla bidens (Šulc, 1907) = Psylla jiangli Yang & Li, 1981, syn. nov.; Cacopsylla jukyungi (Kwon) = C. cinereosignata Luo et al., syn. nov.; Cacopsylla maculatili Li = C. qiuzili Li, syn. nov.; Cacopsylla nigella (Konovalova), comb. nov. from Psylla. The synonymy of P. obongsana Kwon with C. sandolbaea is confirmed.

  18. Ohmic Treatment of Pear Purées (cv. ‘Conference’ in Terms of Some Quality Related Attributes

    Directory of Open Access Journals (Sweden)

    Oana Viorela NISTOR

    2015-06-01

    Full Text Available The effect of ohmic treatment on some quality related characteristics of pear purée (cv. ‘Conference’ such as color, reducing sugars, total phenols, rheological behavior and microbial counts, was analyzed. The inactivation kinetics of pectin methyl esterase (PME in pear crude extract and purée were studied by conventional thermal and ohmic treatments. Thermal inactivation of PME in crude extract was described by a first-order kinetic model. The activation energy values suggested the presence of two isoenzymes with different thermostability. The ohmic heating reduced PME activity by 96% at 25 V·cm-1. Minimal changes induced by ohmic heating on above quality related aspects were observed. Supporting this statement, there were no significant changes in the nutritional and sensorial attributes. It was reported an increase of 3% of reducing sugar content for the ohmic heated samples. The phenolic content of the treated samples registered a reduction of 59% in comparison with fresh pear purée. The pear purée presented a non-Newtonian pseudoplastic behaviour. The Ostwald de Waele model was fitted to rheograms and the consistency coefficient (m and flow behavior index (n were determined. Results obtained for the microbial charge were higher in the control samples. Thus, microbial counts showed complete inactivation of yeast and mold at voltage gradient higher than 17.5 V·cm-1.

  19. 78 FR 53051 - Ethyl-2E,4Z-Decadienoate (Pear Ester); Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2013-08-28

    ..., Biopesticides and Pollution Prevention Division (7511P), Office of Pesticide Programs, Environmental Protection...-decadienoate (pear ester) is a naturally occurring, volatile substance emitted from mature, ripening fruit, that is attractive to the codling moth (CM), Cydia pomonella, a major agricultural pest of pome fruit...

  20. Betalains, Phenols and Antioxidant Capacity in Cactus Pear [Opuntia ficus-indica (L. Mill.] Fruits from Apulia (South Italy Genotypes

    Directory of Open Access Journals (Sweden)

    Clara Albano

    2015-04-01

    Full Text Available Betacyanin (betanin, total phenolics, vitamin C and antioxidant capacity (by Trolox-equivalent antioxidant capacity (TEAC and oxygen radical absorbance capacity (ORAC assays were investigated in two differently colored cactus pear (Opuntia ficus-indica (L. Mill. genotypes, one with purple fruit and the other with orange fruit, from the Salento area, in Apulia (South Italy. In order to quantitate betanin in cactus pear fruit extracts (which is difficult by HPLC because of the presence of two isomers, betanin and isobetanin, and the lack of commercial standard with high purity, betanin was purified from Amaranthus retroflexus inflorescence, characterized by the presence of a single isomer. The purple cactus pear variety showed very high betanin content, with higher levels of phenolics, vitamin C, and antioxidant capacity (TEAC than the orange variety. These findings confirm the potential for exploiting the autochthonous biodiversity of cactus pear fruits. In particular, the purple variety could be an interesting source of colored bioactive compounds which not only have coloring potential, but are also an excellent source of dietary antioxidant components which may have beneficial effects on consumers’ health.

  1. Betalains, Phenols and Antioxidant Capacity in Cactus Pear [Opuntia ficus-indica (L.) Mill.] Fruits from Apulia (South Italy) Genotypes

    Science.gov (United States)

    Albano, Clara; Negro, Carmine; Tommasi, Noemi; Gerardi, Carmela; Mita, Giovanni; Miceli, Antonio; De Bellis, Luigi; Blando, Federica

    2015-01-01

    Betacyanin (betanin), total phenolics, vitamin C and antioxidant capacity (by Trolox-equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assays) were investigated in two differently colored cactus pear (Opuntia ficus-indica (L.) Mill.) genotypes, one with purple fruit and the other with orange fruit, from the Salento area, in Apulia (South Italy). In order to quantitate betanin in cactus pear fruit extracts (which is difficult by HPLC because of the presence of two isomers, betanin and isobetanin, and the lack of commercial standard with high purity), betanin was purified from Amaranthus retroflexus inflorescence, characterized by the presence of a single isomer. The purple cactus pear variety showed very high betanin content, with higher levels of phenolics, vitamin C, and antioxidant capacity (TEAC) than the orange variety. These findings confirm the potential for exploiting the autochthonous biodiversity of cactus pear fruits. In particular, the purple variety could be an interesting source of colored bioactive compounds which not only have coloring potential, but are also an excellent source of dietary antioxidant components which may have beneficial effects on consumers’ health. PMID:26783704

  2. Betalains, Phenols and Antioxidant Capacity in Cactus Pear [Opuntia ficus-indica (L.) Mill.] Fruits from Apulia (South Italy) Genotypes.

    Science.gov (United States)

    Albano, Clara; Negro, Carmine; Tommasi, Noemi; Gerardi, Carmela; Mita, Giovanni; Miceli, Antonio; De Bellis, Luigi; Blando, Federica

    2015-04-01

    Betacyanin (betanin), total phenolics, vitamin C and antioxidant capacity (by Trolox-equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assays) were investigated in two differently colored cactus pear (Opuntia ficus-indica (L.) Mill.) genotypes, one with purple fruit and the other with orange fruit, from the Salento area, in Apulia (South Italy). In order to quantitate betanin in cactus pear fruit extracts (which is difficult by HPLC because of the presence of two isomers, betanin and isobetanin, and the lack of commercial standard with high purity), betanin was purified from Amaranthus retroflexus inflorescence, characterized by the presence of a single isomer. The purple cactus pear variety showed very high betanin content, with higher levels of phenolics, vitamin C, and antioxidant capacity (TEAC) than the orange variety. These findings confirm the potential for exploiting the autochthonous biodiversity of cactus pear fruits. In particular, the purple variety could be an interesting source of colored bioactive compounds which not only have coloring potential, but are also an excellent source of dietary antioxidant components which may have beneficial effects on consumers' health.

  3. Fibulorhizoctonia psychrophila is the causal agent of lenticel spot on apple and pear fruit in the Netherlands

    NARCIS (Netherlands)

    Wenneker, M.; Pham, K.T.K.; Lemmers, M.E.C.; Boer, de F.A.; Leeuwen, van P.J.; Hollinger, T.C.; Geijn, van de F.G.; Thomma, B.P.H.J.

    2016-01-01

    In a survey for postharvest diseases of apples and pears in the Netherlands, an unknown postharvest fruit rot was observed. The disease appeared to originate from infected lenticels. A fungus was consistently isolated from the decayed fruits. The fungal pathogen was isolated on potato dextrose

  4. Effect of polyvinyl alcohol on in vitro rooting capacity of shoots in pear clones (Pyrus communis L.) of different ploidy

    Science.gov (United States)

    Poor adventitious root formation is a major obstacle in micropropagation. In this study, intense efforts have been made for improvement of rooting procedures for triploid, tetraploid, and mixploid clones of the pear cultivar, 'Fertility', obtained by in vitro colchicine treatment. An efficient roo...

  5. Ascorbic acid and tissue browning in pears (Pyrus communis L. cvs Rocha and Conference) under controlled atmosphere conditions

    NARCIS (Netherlands)

    Veltman, R.H.; Kho, R.M.; Schaik, van A.C.R.; Sanders, M.G.; Oosterhaven, J.

    2000-01-01

    The relationships between storage gas composition and ascorbic acid (AA) levels, and between AA levels and the development of internal browning, were studied in 'Conference' and 'Rocha' pears (Pyrus communis L.). In both cultivars, AA levels declined under (browning-inducing) controlled atmosphere

  6. Modeling the effects of temperature and relative humidity on gas exchange of prickly pear cactus (Opuntia spp.) stems

    NARCIS (Netherlands)

    Guevara-Arauza, J.C.; Yahia, E.M.; Cedeno, L.; Tijskens, L.M.M.

    2006-01-01

    A model to estimate gas profile of modified atmosphere packaged (MAP) prickly pear cactus stems was developed and calibrated. The model describes the transient gas exchange taking in consideration the effect of temperature (T) and relative humidity (RH) on film permeability (FPgas), respiration rate

  7. Selection of mutants resistant to black spot disease by chronic irradiation of gamma-rays in Japanese pear 'Osanijisseiki'

    International Nuclear Information System (INIS)

    Masuda, Tetsuo; Yoshioka, Toji; Kotobuki, Kazuo; Sanada, Tetsuro; Inoue, Kosuke; Murata, Kenji; Kitagawa, Kenichi; Tabira, Hiroki; Yoshida, Akira

    1997-01-01

    'Osanijisseiki', a self-compatible, spontaneous bud sport of the Japanese pear 'Nijisseiki' is an excellent cultivar with a smooth skin. However, this cultivar is susceptible to Japanese pear black spot disease caused by Alternaria alternata Japanese pear pathotype. To obtain resistant mutants from 'Osanijisseiki', nursery plants of 'Osanijisseiki' have been irradiated chronically with gamma-rays in the Gamma Field of the Institute of Radiation Breeding, NAR, MAFF, since 1986. Screening tests using AK toxin, a host-specific toxin produced by A. alternata Japanese pear pathotype, were performed form 1988 to 1993. Four branches of young trees planted at a distance of 40 m from the 60 Co source were selected as being resistant mutants in 1991 (IRB 502-13T and IRB 502-14T) and 1993 (IRB 502-17T and IRB 502-18T). Sensitivity of the four resistant mutants to AK-toxin and susceptibility to the pathogen were compared with other of susceptible and resistant cultivars. The results showed that these four mutants possessed intermediate resistance. Furthermore, a mutant, IRB 502-13T, had the same characteristics as the original 'Osanijisseiki', except for the difference in toxin sensitivity. The characteristics of the other mutants, IRB 502 14-T, IRB 502-17T, and IRB 502-18T, care being examined. (author)

  8. Highly efficient amplification of chronic wasting disease agent by protein misfolding cyclical amplification with beads (PMCAb)

    Science.gov (United States)

    Johnson, Chad J.; Aiken, Judd M.; McKenzie, Debbie; Samuel, Michael D.; Pedersen, Joel A.

    2012-01-01

    Protein misfolding cyclic amplification (PMCA) has emerged as an important technique for detecting low levels of pathogenic prion protein in biological samples. The method exploits the ability of the pathogenic prion protein to convert the normal prion protein to a proteinase K-resistant conformation. Inclusion of Teflon® beads in the PMCA reaction (PMCAb) has been previously shown to increase the sensitivity and robustness of detection for the 263 K and SSLOW strains of hamster-adapted prions. Here, we demonstrate that PMCAb with saponin dramatically increases the sensitivity of detection for chronic wasting disease (CWD) agent without compromising the specificity of the assay (i.e., no false positive results). Addition of Teflon® beads increased the robustness of the PMCA reaction, resulting in a decrease in the variability of PMCA results. Three rounds of serial PMCAb allowed detection of CWD agent from a 6.7×10−13 dilution of 10% brain homogenate (1.3 fg of source brain). Titration of the same brain homogenate in transgenic mice expressing cervid prion protein (Tg(CerPrP)1536+/−mice) allowed detection of CWD agent from the 10−6 dilution of 10% brain homogenate. PMCAb is, thus, more sensitive than bioassay in transgenic mice by a factor exceeding 105. Additionally, we are able to amplify CWD agent from brain tissue and lymph nodes of CWD-positive white-tailed deer having Prnp alleles associated with reduced disease susceptibility.

  9. Highly efficient amplification of chronic wasting disease agent by protein misfolding cyclic amplification with beads (PMCAb.

    Directory of Open Access Journals (Sweden)

    Chad J Johnson

    Full Text Available Protein misfolding cyclic amplification (PMCA has emerged as an important technique for detecting low levels of pathogenic prion protein in biological samples. The method exploits the ability of the pathogenic prion protein to convert the normal prion protein to a proteinase K-resistant conformation. Inclusion of Teflon® beads in the PMCA reaction (PMCAb has been previously shown to increase the sensitivity and robustness of detection for the 263 K and SSLOW strains of hamster-adapted prions. Here, we demonstrate that PMCAb with saponin dramatically increases the sensitivity of detection for chronic wasting disease (CWD agent without compromising the specificity of the assay (i.e., no false positive results. Addition of Teflon® beads increased the robustness of the PMCA reaction, resulting in a decrease in the variability of PMCA results. Three rounds of serial PMCAb allowed detection of CWD agent from a 6.7 × 10(-13 dilution of 10% brain homogenate (1.3 fg of source brain. Titration of the same brain homogenate in transgenic mice expressing cervid prion protein (Tg(CerPrP1536(+/- mice allowed detection of CWD agent from the 10(-6 dilution of 10% brain homogenate. PMCAb is, thus, more sensitive than bioassay in transgenic mice by a factor exceeding 10(5. Additionally, we are able to amplify CWD agent from brain tissue and lymph nodes of CWD-positive white-tailed deer having Prnp alleles associated with reduced disease susceptibility.

  10. Ultrasmall volume molecular isothermal amplification in microfluidic chip with advanced surface processing

    International Nuclear Information System (INIS)

    Huang Guoliang; Yang Xiaoyong; Ma Li; Yang Xu

    2011-01-01

    In this paper, we developed a metal micro-fluidic chip with advanced surface processing for ultra-small volume molecular isothermal amplification. This method takes advantages of the nucleic acid amplification with good stability and consistency, high sensitivity about 31 genomic DNA copies and bacteria specific gene identification. Based on the advanced surface processing, the bioreaction assays of nucleic acid amplification was dropped about 392nl in volume. A high numerical aperture confocal optical detection system was advanced to sensitively monitor the DNA amplification with low noise and high power collecting fluorescence near to the optical diffraction limit. A speedy nucleic acid isothermal amplification was performed in the ultra-small volume microfluidic chip, where the time at the inflexions of second derivative to DNA exponential amplified curves was brought forward and the sensitivity was improved about 65 folds to that of in current 25μl Ep-tube amplified reaction, which indicates a promising clinic molecular diagnostics in the droplet amplification.

  11. Real-time electrochemical monitoring of isothermal helicase-dependent amplification of nucleic acids.

    Science.gov (United States)

    Kivlehan, Francine; Mavré, François; Talini, Luc; Limoges, Benoît; Marchal, Damien

    2011-09-21

    We described an electrochemical method to monitor in real-time the isothermal helicase-dependent amplification of nucleic acids. The principle of detection is simple and well-adapted to the development of portable, easy-to-use and inexpensive nucleic acids detection technologies. It consists of monitoring a decrease in the electrochemical current response of a reporter DNA intercalating redox probe during the isothermal DNA amplification. The method offers the possibility to quantitatively analyze target nucleic acids in less than one hour at a single constant temperature, and to perform at the end of the isothermal amplification a DNA melt curve analysis for differentiating between specific and non-specific amplifications. To illustrate the potentialities of this approach for the development of a simple, robust and low-cost instrument with high throughput capability, the method was validated with an electrochemical system capable of monitoring up to 48 real-time isothermal HDA reactions simultaneously in a disposable microplate consisting of 48-electrochemical microwells. Results obtained with this approach are comparable to that obtained with a well-established but more sophisticated and expensive fluorescence-based method. This makes for a promising alternative detection method not only for real-time isothermal helicase-dependent amplification of nucleic acid, but also for other isothermal DNA amplification strategies.

  12. Simple system for isothermal DNA amplification coupled to lateral flow detection.

    Directory of Open Access Journals (Sweden)

    Kristina Roskos

    Full Text Available Infectious disease diagnosis in point-of-care settings can be greatly improved through integrated, automated nucleic acid testing devices. We have developed an early prototype for a low-cost system which executes isothermal DNA amplification coupled to nucleic acid lateral flow (NALF detection in a mesofluidic cartridge attached to a portable instrument. Fluid handling inside the cartridge is facilitated through one-way passive valves, flexible pouches, and electrolysis-driven pumps, which promotes a compact and inexpensive instrument design. The closed-system disposable prevents workspace amplicon contamination. The cartridge design is based on standard scalable manufacturing techniques such as injection molding. Nucleic acid amplification occurs in a two-layer pouch that enables efficient heat transfer. We have demonstrated as proof of principle the amplification and detection of Mycobacterium tuberculosis (M.tb genomic DNA in the cartridge, using either Loop Mediated Amplification (LAMP or the Exponential Amplification Reaction (EXPAR, both coupled to NALF detection. We envision that a refined version of this cartridge, including upstream sample preparation coupled to amplification and detection, will enable fully-automated sample-in to answer-out infectious disease diagnosis in primary care settings of low-resource countries with high disease burden.

  13. Morphological, physiological and biochemical responses to soil water deficit in seedlings of three populations of wild pear tree (Pyrus boisseriana

    Directory of Open Access Journals (Sweden)

    Zarafshar, M.

    2014-01-01

    Full Text Available Water shortage limits the production of fruit orchards, such as pear, in arid and semi-arid regions. The identification of wild pear germplasm for potential use as rootstock would be valuable for pear cultivation in semi-arid regions. The relative drought tolerance of wild pear germplasm (Pyrus boisseriana from three different populations distributed along an elevational gradient ('semi-arid 1,000', 'semi-wet 1,350' and 'semi-wet 1,600' populations was evaluated in a greenhouse trial. Established container-grown seedlings were exposed to 18 days of simulated drought, or not, followed by a seven day recovery period. Biomass allocation and accumulation, physiological (stomatal conductance, photosynthesis, transpiration, xylem water potential and biochemical parameters (leaf pigments, free proline, malondialdehyde and hydrogen peroxide production were evaluated. Although all populations were able to recover from water shortage, thereby proving to be relatively drought tolerant, some differences between populations were detected for gas exchange parameters, biomass accumulation and proline concentration in favor of the 'semi-arid 1,000' elevation population, which was more drought tolerant. This population showed the most rapid and complete recovery of physiological activity (stomatal conductance and carbon fixation. In addition, all populations showed an increase in carotenoid content in the leaves. Overall, we showed that plants from the 'semi-arid 1,000' elevation had greater tolerance to drought than those from the higher elevations (semi-wet populations. It therefore appears that plants from the 'semi-arid 1,000' elevation represent a promising source of material to be tested as rootstock for commercial scions of pear in field conditions in areas prone to suffer from water deficit.

  14. Exposure to minimally processed pear and melon during shelf life could modify the pathogenic potential of Listeria monocytogenes.

    Science.gov (United States)

    Colás-Medà, Pilar; Viñas, Inmaculada; Oliveira, Márcia; Anguera, Marina; Serrano, Jose C E; Abadias, Maribel

    2017-04-01

    Survival and virulence of foodborne pathogens can be influenced by environmental factors such as the intrinsic properties of food as well as the extrinsic properties that contribute to food shelf life (e.g., temperature and gas atmosphere). The direct contribution of food matrix characteristics on the survival of L. monocytogenes during fresh-cut fruit shelf life is not very well understood. In addition, the gastrointestinal tract is the primary route of listeriosis infection and penetration of the intestinal epithelial cell barrier is the first step in the infection process. Hence, the pathogenic potential of L. monocytogenes, measured as the capability for the organism to survive a simulated gastrointestinal tract and the proportion of cells able to subsequently adhere to and invade differentiated Caco-2 cells, subjected to fresh-cut pear and melon shelf life, was investigated. Samples were inoculated, stored at 10 °C for 7 days and evaluated after inoculation and again after 2 and 7 days of storage. A decrease in L. monocytogenes' capacity to survive a simulated gastrointestinal tract was observed with increasing storage time, regardless of the fruit matrix evaluated. Furthermore, L. monocytogenes placed on fresh-cut pear and melon was subjected to an attachment and invasion assay after crossing the simulated gastrointestinal tract. After inoculation, pathogen on fresh-cut pear showed 5-fold more capacity to adhere to Caco-2 cells than pathogen on fresh-cut melon. After 2 days of storage, L. monocytogenes grown on fresh-cut melon showed similar adhesive capacity (1.11%) than cells grown on pear (1.83%), but cells grown on melon had the higher invasive capacity (0.0093%). We can conclude that minimally processed melon could represent a more important hazard than pear under the studied shelf life. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Investigation the Frost Resistance of Vegetative and Reproductive Buds of Pear Cultivars in Mashhad Climate Condition

    Directory of Open Access Journals (Sweden)

    shadan khorshidi

    2017-09-01

    Full Text Available Introduction: Most deciduous trees need low temperature to break flower bud dormancy. One of the most important abiotic stresses is low temperature which limits production of temperate fruits. Pear production has been considerably reduced in recent years. Important pear cultivars show different levels of resistance to cold. Cold compatibility followed by resistance increase is controlled genetically and contains several mechanisms which lead to production of different metabolites such as: polypeptides, amino acids and sugars. The object of this research was to evaluate the frost resistance of different ‘Dare Gazi’ genotypes and other pear cultivars in Mashhad climate condition. Materials and Methods: This study was conducted to investigate the frost resistance of 23 ‘Dare Gazi’ pear genotypes and nine other cultivars include: ‘William’s’, ‘Bell de june’, ‘Spadona’, ‘Koshia’, ‘Domkaj’, ‘Torsh’, ‘Sebri’ and ‘Tabrizi’. Plant material contained vegetative and reproductive buds of one-year-old shoot samples which were collected from 25-year old trees on March 2014, four days after winter cold (-6.6 °C in three directions of trees and sent to the laboratory. Frost damages of vegetative and reproductive buds were investigated based on visual observations (%, electrolyte leakage (EC and proline content. EC was measured with a Metrohm 644 digital conductivity meter and proline content was measured based on Bates et al. (1973 method, using acid ninhydrin. The experiment was performed on completely randomized experimental design with three replications. Statistical analysis was carried out using MSTAT-C and Excel software. Mean values were compared using the least significance difference test (LSD at 1% levels. Cluster analysis was conducted by SPSS 16 program. Results and Discussion: Highest EC of reproductive buds was observed in ‘Dare Gazi’ 10, 19, ‘Tabrizi’ and ‘Torsh’ whereas ‘Dare Gazi’ 8, 18

  16. Rapid and Sensitive Isothermal Detection of Nucleic-acid Sequence by Multiple Cross Displacement Amplification.

    Science.gov (United States)

    Wang, Yi; Wang, Yan; Ma, Ai-Jing; Li, Dong-Xun; Luo, Li-Juan; Liu, Dong-Xin; Jin, Dong; Liu, Kai; Ye, Chang-Yun

    2015-07-08

    We have devised a novel amplification strategy based on isothermal strand-displacement polymerization reaction, which was termed multiple cross displacement amplification (MCDA). The approach employed a set of ten specially designed primers spanning ten distinct regions of target sequence and was preceded at a constant temperature (61-65 °C). At the assay temperature, the double-stranded DNAs were at dynamic reaction environment of primer-template hybrid, thus the high concentration of primers annealed to the template strands without a denaturing step to initiate the synthesis. For the subsequent isothermal amplification step, a series of primer binding and extension events yielded several single-stranded DNAs and single-stranded single stem-loop DNA structures. Then, these DNA products enabled the strand-displacement reaction to enter into the exponential amplification. Three mainstream methods, including colorimetric indicators, agarose gel electrophoresis and real-time turbidity, were selected for monitoring the MCDA reaction. Moreover, the practical application of the MCDA assay was successfully evaluated by detecting the target pathogen nucleic acid in pork samples, which offered advantages on quick results, modest equipment requirements, easiness in operation, and high specificity and sensitivity. Here we expounded the basic MCDA mechanism and also provided details on an alternative (Single-MCDA assay, S-MCDA) to MCDA technique.

  17. Loop-mediated isothermal amplification (LAMP) shield for Arduino DNA detection

    NARCIS (Netherlands)

    Velders, Aldrik H.; Schoen, Cor; Saggiomo, Vittorio

    2018-01-01

    Objective: Loop-mediated isothermal amplification (LAMP) of DNA is gaining relevance as a method to detect nucleic acids, as it is easier, faster, and more powerful than conventional Polymerase Chain Reaction. However, LAMP is still mostly used in laboratory settings, because of the lack of a cheap

  18. Amplification of Chirality through Self-Replication of Micellar Aggregates in Water

    KAUST Repository

    Bukhriakov, Konstantin

    2015-03-17

    We describe a system in which the self-replication of micellar aggregates results in a spontaneous amplification of chirality in the reaction products. In this system, amphiphiles are synthesized from two "clickable" fragments: a water-soluble "head" and a hydrophobic "tail". Under biphasic conditions, the reaction is autocatalytic, as aggregates facilitate the transfer of hydrophobic molecules to the aqueous phase. When chiral, partially enantioenriched surfactant heads are used, a strong nonlinear induction of chirality in the reaction products is observed. Preseeding the reaction mixture with an amphiphile of one chirality results in the amplification of this product and therefore information transfer between generations of self-replicating aggregates. Because our amphiphiles are capable of catalysis, information transfer, and self-assembly into bounded structures, they present a plausible model for prenucleic acid "lipid world" entities. © 2015 American Chemical Society.

  19. Comparison of the influence of defined storing conditions on asian and european pear cultivars

    Directory of Open Access Journals (Sweden)

    Petr Šnurkovič

    2016-12-01

    Full Text Available The study observed changes in the material composition of European (Conference and Asian (Yali pears during the storage period at different temperatures. Three different temperatures were selected for storing, i.e. 1 °C, 5 °C and 20 °C. Assessed for each of the pieces of fruit were flesh firmness, titratable acidity, soluble solids content, content of organic acids, and production of ethylene and carbon dioxide. Fruit stored at 1 °C and 5 °C was analysed before the moment of putting to the store and then after 25, 55 and 70 days of storing. Fruit stored at 20 °C was analysed before the moment of putting to the store and then after 14, 22 and 30 days of storing. The respiration intensity observed through carbon dioxide production at refrigeration conditions was approximately of the same progress for both of the varieties. For the Yali variety, the intensity of respiration of the fruit at the start of the storing period strongly decreased.The same progress was recorded for the Conference variety. Storing at 20 °C increased the respiration intensity. The varieties Yali and Conference which were stored under the temperature of 5 °C had the highest CO2 production after 70 days of storage. For both varieties the lowest ethylene production during storage was observed at 1 °C. Ethylene production was higher in Yali pear fruits. The Yali variety stored at 20 °C from the beginning produced up to 10 times higher concentration of ethylene than the fruit of the Conference variety. The highest amount of ethylen by the Conference variety was produced by the fruits which were stored under the temperature of 5 °C. At the beginning of the storage period the Conference pears had two-fold higher flesh firmness than the Yali fruit. Fruits of the varieties Yali and Conference which were stored 70 days under the temperature of 1 °C had the highest flesh firmness. For soluble solids and titratable acidity no clear progress was recorded. Malic acid was

  20. THE LINE FOR PRODUCTION OF DRIED APPLES, PEARS, CARROTS, PUMPKIN AND CHIPS

    Directory of Open Access Journals (Sweden)

    G. V. Kalashnikov

    2015-01-01

    Full Text Available The line is intended for processing of fruit and vegetable raw materials and receiving dried apples, pears, carrots, pumpkins and the fruit-and-vegetable of chips. The line solves problems of improvement of quality of a ready-made product and thermal production efficiency due to more rational alternation of the technological modes of a moisture increment and dehumidification with high extent of use of an energy potential of the heat carrier, use of the inert heat carrier (steam identical by the form for technological thermal processes, decrease in specific energy consumption and metal consumption, and also an intensification of moisture evaporation and creation of the compact multipurpose technological line for production of fruit and vegetable products with the expanded range. The technological production line of dried apples, pears, carrots, pumpkin and fruit and vegetable chips contains the jet washer, the inspection conveyor, the size grader, the car for removal of a seed nest and the device are sharp fruits and vegetables on plates, the sulfiter, the dryer and the packing automatic packing machine. Thus the line contains the combined toroidal device for heatmoisture of handling continuous action divided into sections: section of heating of raw materials, section of convective drying, section of preliminary hydration, which is located between microwave drying sections, and the section of cooling of the dried-up product intended for bringing a product to final readiness. The equipment complex from the drum car with the washing block and multipurpose installation with crushing of raw materials and office of sunflower seeds taking into account raw materials type is provided in lines. Are used recirculation a contour, the heating of the initial raw material fulfilled after drying of pairs and a condensate in the closed contour for creation energy-saving of the "know-how" of a ready product. The line represents modular blocks and is recustomized

  1. Internal amplification control of PCR for the Glu1-Dx5 allele in wheat.

    Science.gov (United States)

    Heim, H N; Vieira, E S N; Polo, L R T; Lima, N K; Silva, G J; Linde, G A; Colauto, N B; Schuster, I

    2017-08-17

    One of the limiting factors in using dominant markers is the unique amplification of the target fragment. Therefore, failures in polymerase chain reaction (PCR) or non-amplifications can be interpreted as an absence of the allele. The possibility of false negatives implies in reduced efficiency in the selection process in genetic breeding programs besides the loss of valuable genetic material. Thus, this study aimed to evaluate the viability of a microsatellite marker as an internal amplification control with a dominant marker for the wheat Glu1-Dx5 gene. A population of 77 wheat cultivars/breeding lines was analyzed. Fourteen microsatellite markers were analyzed in silico regarding the formation of dimers and clamps. The biplex reaction conditions were optimized, and the Xbarc117 marker was selected as the internal amplification control with a Glu1-Dx5 marker in wheat. It was concluded that the Xbarc117 microsatellite marker was effective in the simultaneous amplification with a dominant Glu1-Dx5 marker, making biplex PCR viable in wheat for the studied markers.

  2. A novel steric effect-regulated isothermal exponential amplification technology for the one-step homogeneous sensing of proteins.

    Science.gov (United States)

    Wu, Wanghua; Pan, Wufan; Yu, Dongdong; Yuan, Zhen; Qin, Yazhou; Lu, Yuxiang; Zhang, Tao; Zhou, Jianguang

    2018-02-12

    A simple and homogeneous technology, the steric effect-regulated isothermal exponential amplification reaction (SER-EXPAR), was developed to sense proteins. By using a small molecule linked DNA nanostructure, termed enzyme-binding hairpin (EBH), the protein-small molecule binding events could be readily sensed by utilizing the steric effect generated between the protein and enzyme. It set free the enzyme to be active again, thus regulating the amplification rate of EXPAR.

  3. Privacy amplification for quantum key distribution

    International Nuclear Information System (INIS)

    Watanabe, Yodai

    2007-01-01

    This paper examines classical privacy amplification using a universal family of hash functions. In quantum key distribution, the adversary's measurement can wait until the choice of hash functions is announced, and so the adversary's information may depend on the choice. Therefore the existing result on classical privacy amplification, which assumes the independence of the choice from the other random variables, is not applicable to this case. This paper provides a security proof of privacy amplification which is valid even when the adversary's information may depend on the choice of hash functions. The compression rate of the proposed privacy amplification can be taken to be the same as that of the existing one with an exponentially small loss in secrecy of a final key. (fast track communication)

  4. Multiplex amplification of large sets of human exons.

    Science.gov (United States)

    Porreca, Gregory J; Zhang, Kun; Li, Jin Billy; Xie, Bin; Austin, Derek; Vassallo, Sara L; LeProust, Emily M; Peck, Bill J; Emig, Christopher J; Dahl, Fredrik; Gao, Yuan; Church, George M; Shendure, Jay

    2007-11-01

    A new generation of technologies is poised to reduce DNA sequencing costs by several orders of magnitude. But our ability to fully leverage the power of these technologies is crippled by the absence of suitable 'front-end' methods for isolating complex subsets of a mammalian genome at a scale that matches the throughput at which these platforms will routinely operate. We show that targeting oligonucleotides released from programmable microarrays can be used to capture and amplify approximately 10,000 human exons in a single multiplex reaction. Additionally, we show integration of this protocol with ultra-high-throughput sequencing for targeted variation discovery. Although the multiplex capture reaction is highly specific, we found that nonuniform capture is a key issue that will need to be resolved by additional optimization. We anticipate that highly multiplexed methods for targeted amplification will enable the comprehensive resequencing of human exons at a fraction of the cost of whole-genome resequencing.

  5. Micropropagation of Pear Rootstock (Pyrus Communis) by using tissue culture technique and gamma irradiation

    International Nuclear Information System (INIS)

    El-Sharnouby, M.E.; ESSAM, E.R.; Ayoub, S.

    2006-01-01

    New growing shoots from healthy pear rootstock (Pyrus communis) trees were taken and sterilized 3 times in dipping water. Explants were subjected to antioxidant treatment, different media, different additives and different BAP and NAA concentrations. The obtained results showed that Murashig-Skoog (MS) supplemented with 1 mg/l BA was better than Gamborg medium. Adding antioxidant solution and adenine sulphate to the culture medium was preferred for maximizing explants development. Exposing the explants to gamma irradiation at different doses decreased tissue culture parameters with increasing gamma doses. However, the low dose of gamma rays (1 Krad) significantly increased the number of shoots than other gamma treatments. Adding of BAP at 2 mg/l to the culture medium increased number and length of shoots. However, addition of 1 mg/l NAA to the rooting medium led to increase the root formation

  6. Studies of the shapes of heavy pear-shaped nuclei at ISOLDE

    Energy Technology Data Exchange (ETDEWEB)

    Butler, P. A., E-mail: peter.butler@liverpool.ac.uk [Oliver Lodge Laboratory, University of Liverpool, Liverpool L69 7ZE (United Kingdom)

    2016-07-07

    For certain combinations of protons and neutrons there is a theoretical expectation that the shape of nuclei can assume octupole deformation, which would give rise to reflection asymmetry or a ”pear-shape” in the intrinsic frame, either dynamically (octupole vibrations) or statically (permanent octupole deformation). I will briefly review the historic evidence for reflection asymmetry in nuclei and describe how recent experiments carried out at REX-ISOLDE have constrained nuclear theory and how they contribute to tests of extensions of the Standard Model. I will also discuss future prospects for measuring nuclear shapes from Coulomb Excitation: experiments are being planned that will exploit beams from HIE-ISOLDE that are cooled in the TSR storage ring and injected into a solenoidal spectrometer similar to the HELIOS device developed at the Argonne National Laboratory.

  7. Chemical composition and antibacterial activity of Opuntia ficus-indica f. inermis (cactus pear) flowers.

    Science.gov (United States)

    Ennouri, Monia; Ammar, Imene; Khemakhem, Bassem; Attia, Hamadi

    2014-08-01

    Opuntia ficus-indica f. inermis (cactus pear) flowers have wide application in folk medicine. However, there are few reports focusing on their biological activity and were no reports on their chemical composition. The nutrient composition and hexane extracts of Opuntia flowers at 4 flowering stages and their antibacterial and antifungal activities were investigated. The chemical composition showed considerable amounts of fiber, protein, and minerals. Potassium (K) was the predominant mineral followed by calcium (Ca), magnesium (Mg), sodium (Na), iron (Fe), and zinc (Zn). The main compounds in the various hexane extracts were 9.12-octadecadienoic acid (29-44%) and hexadecanoic acid (8.6-32%). The antibacterial activity tests showed that O. inermis hexane extracts have high effectiveness against Escherichia coli and Staphylococcus aureus, making this botanical source a potential contender as a food preservative or food control additive.

  8. Determination of some mineral contents of prickly pear (Opuntia ficus-indica L.) seed flours.

    Science.gov (United States)

    Al-Juhaimi, Fahad; Özcan, Mehmet Musa

    2013-05-01

    The aim of this study was to determine some mineral contents of prickly pear (Opuntia fıcus-indica L.) seeds collected from different locations. The mineral contents of seeds were established by inductively coupled plasma atomic emission spectrometry. All the seeds contained Ca, K, Mg and P at high levels. Calcium content ranged between 268.5 (sample no. 11) and 674.8 ppm (sample no. 4). The level of K changed between 346.7 (sample no. 1) and 676.1 ppm (sample no. 13). Phosphorus content of seeds varied between 1,173.6 (sample no. 14) and 1,871.3 ppm (sample no. 1). It is apparent that seeds are good sources of the macro and micro minerals and can be consumed as a food ingredient to provide nutrition.

  9. Improving the quality of fresh-cut apples, pears, and melons using natural additives.

    Science.gov (United States)

    Alandes, L; Quiles, A; Pérez-Munuera, I; Hernando, I

    2009-03-01

    Improving the quality of different fresh-cut fruits by adding natural substances was studied. "Fuji" apples, "Flor de Invierno" pears, and "Piel de Sapo" melons were treated with calcium lactate, N-acetyl-L-cysteine, glutathione, and malic acid and stored for 4 wk at 4 degrees C. Instrumental texture (penetration), microstructure (light microscopy), acidity, soluble solids, color, pectinmethylesterase activity, and microflora were studied. The results showed that the combined treatment reinforced the cell walls strengthening the structure and texture of these fruits and maintained the L* and a* values throughout 4 wk of storage at 4 degrees C. The combination of additives provided low microbial counts in apples until the 4th week and in melons until the 2nd week. So, this combined treatment could be used to extend the shelf life of some fresh-cut fruits while preserving their quality.

  10. Refillable and magnetically actuated drug delivery system using pear-shaped viscoelastic membrane

    KAUST Repository

    So, Hongyun

    2014-07-01

    We report a refillable and valveless drug delivery device actuated by an external magnetic field for on-demand drug release to treat localized diseases. The device features a pear-shaped viscoelastic magnetic membrane inducing asymmetrical deflection and consecutive touchdown motion to the bottom of the dome-shaped drug reservoir in response to a magnetic field, thus achieving controlled discharge of the drug. Maximum drug release with 18 ± 1.5 μg per actuation was achieved under a 500 mT magnetic flux density, and various controlled drug doses were investigated with the combination of the number of accumulated actuations and the strength of the magnetic field.

  11. Rolling circle amplification of metazoan mitochondrialgenomes

    Energy Technology Data Exchange (ETDEWEB)

    Simison, W. Brian; Lindberg, D.R.; Boore, J.L.

    2005-07-31

    Here we report the successful use of rolling circle amplification (RCA) for the amplification of complete metazoan mt genomes to make a product that is amenable to high-throughput genome sequencing techniques. The benefits of RCA over PCR are many and with further development and refinement of RCA, the sequencing of organellar genomics will require far less time and effort than current long PCR approaches.

  12. MORPHOLOGICAL CHARACTERISTICS AND FORAGE PRODUCTIVITY OF IRRIGATED CACTUS PEAR UNDER DIFFERENT CUTTING INTENSITIES

    Directory of Open Access Journals (Sweden)

    GUILHERME FERREIRA DA COSTA LIMA

    2016-01-01

    Full Text Available This study evaluated the effect of different cutting intensities and years of harvesting on the morphological characteristics and production of fresh (FMP and dry matter (DMP of cactus pear cv. Gigante (Opuntia ficus-indica Mill under conditions of irrigation, high planting density and fertilization, with 12 months of regrowth. The experimental was completely randomized in a factorial design (3 × 2 with 12 replicates. The treatments were three cutting intensities (preserving the mother cladode (PMC, primary cladodes (PPC, or secondary cladodes (PSC, and two years of harvesting. The soil was classified as Cambisol Haplicum and the irrigation water was classified as C4S1 (EC 5.25 dS.m-1 density of 50,000 plants ha-1. The research evaluated plant height, number of cladodes per plant (NCP, length, width, perimeter and thickness of the cladodes, cladode area (CA, cladode area index (CAI, FMP and DMP. There was no significant interaction between treatments (P > 0.05 for the variables plant height, NCP, CAI and FMP. The variables related to cladode morphology showed a significant interaction (P < 0.05. The treatment PSC resulted in a greater DMP (P < 0.05 with a mean of 27.17 Mg ha-1 yr-1, compared to PPC (18.58 Mg ha-1 yr-1 or PMC (11.78 Mg ha-1 yr-1. The treatment PSC promoted greater NCP and forage productivity at harvest and can be considered as a management practice for the sustainability of cactus pear cv. Gigante under irrigation. The more important morphological characteristics were also influenced by the lower cutting intensities.

  13. Estimation of loss of genetic diversity in modern Japanese cultivars by comparison of diverse genetic resources in Asian pear (Pyrus spp.).

    Science.gov (United States)

    Nishio, Sogo; Takada, Norio; Saito, Toshihiro; Yamamoto, Toshiya; Iketani, Hiroyuki

    2016-06-14

    Pears (Pyrus spp.) are one of the most important fruit crops in temperate regions. Japanese pear breeding has been carried out for over 100 years, working to release new cultivars that have good fruit quality and other desirable traits. Local cultivar 'Nijisseiki' and its relatives, which have excellent fruit texture, have been repeatedly used as parents in the breeding program. This strategy has led to inbreeding within recent cultivars and selections. To avoid inbreeding depression, we need to clarify the degree of inbreeding among crossbred cultivars and to introgress genetic resources that are genetically different from modern cultivars and selections. The objective of the present study was to clarify the genetic relatedness between modern Japanese pear cultivars and diverse Asian pear genetic resources. We genotyped 207 diverse accessions by using 19 simple sequence repeat (SSR) markers. The heterozygosity and allelic richness of modern cultivars was obviously decreased compared with that of wild individuals, Chinese pear cultivars, and local cultivars. In analyses using Structure software, the 207 accessions were classified into four clusters (K = 4): one consisting primarily of wild individuals, one of Chinese pear cultivars, one of local cultivars from outside the Kanto region, and one containing both local cultivars from the Kanto region and crossbred cultivars. The results of principal coordinate analysis (PCoA) were similar to those from the Structure analysis. Wild individuals and Chinese pears appeared to be distinct from other groups, and crossbred cultivars became closer to 'Nijisseiki' as the year of release became more recent. Both Structure and PCoA results suggest that the modern Japanese pear cultivars are genetically close to local cultivars that originated in the Kanto region, and that the genotypes of the modern cultivars were markedly biased toward 'Nijisseiki'. Introgression of germplasm from Chinese pear and wild individuals that are

  14. Epidemiological Features and Trends of Brown Spot of Pear Disease Based on the Diversity of Pathogen Populations and Climate Change Effects.

    Science.gov (United States)

    Moragrega, Concepció; Puig, Mireia; Ruz, Lídia; Montesinos, Emilio; Llorente, Isidre

    2018-02-01

    Brown spot of pear, caused by the fungus Stemphylium vesicarium, is an emerging disease of economic importance in several pear-growing areas in Europe. In recent years, new control strategies combining sanitation practices and fungicide applications according to developed forecasting models have been introduced to manage the disease. However, the pathogenic and saprophytic behavior of this pathogen makes it difficult to manage the disease. In addition, climate change can also result in variations in the severity and geographical distribution of the disease. In this study, ecological and epidemiological aspects of brown spot of pear disease related to inoculum characterization and climate change impact were elucidated. The pathogenic variation in S. vesicarium populations from pear orchards and its relationship to inoculum sources (air samples, leaf debris, and infected host and nonhost tissues) was determined using multivariate analysis. In total, six variables related to infection and disease development on cultivar Conference pear detached leaves of 110 S. vesicarium isolates were analyzed. A high proportion of isolates (42%) were nonpathogenic to pear; 85% of these nonpathogenic isolates were recovered from air samples. Most isolates recovered from lesions (93%) and pseudothecia (83%) were pathogenic to pear. A group of pathogenic isolates rapidly infected cultivar Conference pear leaves resulted in disease increase that followed a monomolecular model, whereas some S. vesicarium isolates required a period of time after inoculation to initiate infection and resulted in disease increase that followed a logistic model. The latter group was mainly composed of isolates recovered from pseudothecia on leaf debris, whereas the former group was mainly composed of isolates recovered from lesions on pear fruit and leaves. The relationship between the source of inoculum and pathogenic/aggressiveness profile was confirmed by principal component analysis. The effect of

  15. Molecular Characterization, Evolution, and Expression Profiling of the Dirigent (DIR Family Genes in Chinese White Pear (Pyrus bretschneideri

    Directory of Open Access Journals (Sweden)

    Xi Cheng

    2018-04-01

    Full Text Available Stone cells content and size are the key factors determining the internal quality of the pear fruit. Synthesis of lignin and thickening of secondary cell wall are the keys to the development of stone cells. The polymerization of monolignols and secondary cell wall formation requires the participation of dirigent proteins (DIRs. In recent years, DIR family have been studied in higher plants, but lack of comprehensive study in the pear DIR (PbDIR family. This study focuses on the identification and analysis of PbDIR family for the first time. We identified 35 PbDIRs from the pear genome, 89% of which are intronless genes. Phylogenetic tree and chromosome localization analysis showed that 35 PbDIRs were divided into four subfamilies (DIR-a, -b/d, -e, and -g and irregularly distributed among 10 chromosomes. In addition, we identified 29, 26, and 14 DIRs from the other three Rosids (peach, Mei, and grape, respectively. Interspecies microsynteny analysis revealed the collinear gene pairs between pear and peach are the most. Temporal expression analysis showed that the expression changes of seven PbDIRs (DIR-a subfamily: PbDIR4 and PbDIR5; DIR-b/d subfamily: PbDIR11; DIR-g subfamily: PbDIR19; DIR-e subfamily: PbDIR23, 25 and 26 in fruits were consistent with the changes of fruit lignin and stone cells contents. In addition, the subfamily of PbDIRs in fruits showed significant responses after treatment with ABA, SA, and MeJA. According to the protein tertiary structure, key amino acid residues and expression patterns analysis found that PbDIR4 might be involved in the metabolism of lignin and related to stone cells contents in pear fruits. In this study, we systematically analyzed the structure, evolution, function and expression of PbDIR family, which not only confirmed the characteristics of PbDIR family, but also laid the foundation for revealing the role of DIR in pear stone cell development and lignin polymerization.

  16. Isothermal Recombinase Polymerase amplification (RPA) of Schistosoma haematobium DNA and oligochromatographic lateral flow detection.

    Science.gov (United States)

    Rosser, A; Rollinson, D; Forrest, M; Webster, B L

    2015-09-04

    Accurate diagnosis of urogenital schistosomiasis is vital for surveillance/control programs. Amplification of schistosome DNA in urine by PCR is sensitive and specific but requires infrastructure, financial resources and skilled personnel, often not available in endemic areas. Recombinase Polymerase Amplification (RPA) is an isothermal DNA amplification/detection technology that is simple, rapid, portable and needs few resources. Here a Schistosoma haematobium RPA assay was developed and adapted so that DNA amplicons could be detected using oligochromatographic Lateral Flow (LF) strips. The assay successfully amplified S. haematobium DNA at 30-45 °C in 10 mins and was sensitive to a lower limit of 100 fg of DNA. The assay was also successful with the addition of crude urine, up to 5% of the total reaction volume. Cross amplification occurred with other schistosome species but not with other common urine microorganisms. The LF-RPA assay developed here can amplify and detect low levels of S. haematobium DNA. Reactions are rapid, require low temperatures and positive reactions are interpreted using lateral flow strips, reducing the need for infrastructure and resources. This together with an ability to withstand inhibitors within urine makes RPA a promising technology for further development as a molecular diagnostic tool for urogenital schistosomiasis.

  17. Systematic analysis and comparison of the PHD-Finger gene family in Chinese pear (Pyrus bretschneideri) and its role in fruit development.

    Science.gov (United States)

    Cao, Yunpeng; Han, Yahui; Meng, Dandan; Abdullah, Muhammad; Li, Dahui; Jin, Qing; Lin, Yi; Cai, Yongping

    2018-04-20

    PHD-finger proteins, which belongs to the type of zinc finger family, and that play an important role in the regulation of both transcription and the chromatin state in eukaryotes. Currently, PHD-finger proteins have been well studied in animals, while few studies have been carried out on their function in plants. In the present study, 129 non-redundant PHD-finger genes were identified from 5 Rosaceae species (pear, apple, strawberry, mei, and peach); among them, 31 genes were identified in pear. Subsequently, we carried out a bioinformatics analysis of the PHD-finger genes. Thirty-one PbPHD genes were divided into 7 subfamilies based on the phylogenetic analysis, which are consistent with the intron-exon and conserved motif analyses. In addition, we identified five segmental duplication events, implying that the segmental duplications might be a crucial role in the expansion of the PHD-finger gene family in pear. The microsynteny analysis of five Rosaceae species showed that there were independent duplication events in addition to the genome-wide duplication of the pear genome. Subsequently, ten expressed PHD-finger genes of pear fruit were identified using qRT-PCR, and one of these genes, PbPHD10, was identified as an important candidate gene for the regulation of lignin synthesis. Our research provides useful information for the further analysis of the function of PHD-finger gene family in pear.

  18. A Label-Free and Sensitive Fluorescent Qualitative Assay for Bisphenol A Based on Rolling Circle Amplification/Exonuclease III-Combined Cascade Amplification.

    Science.gov (United States)

    Li, Xia; Song, Juan; Xue, Qing-Wang; You, Fu-Heng; Lu, Xia; Kong, Yan-Cong; Ma, Shu-Yi; Jiang, Wei; Li, Chen-Zhong

    2016-10-21

    Bisphenol A (BPA) detection in drinking water and food packaging materials has attracted much attention since the discovery that BPA can interfere with normal physiological processes and cause adverse health effects. Here, we constructed a label-free aptamer fluorescent assay for selective and sensitive detection of BPA based on the rolling circle amplification (RCA)/Exonuclease III (Exo III)-combined cascade amplification strategy. First, the duplex DNA probe (RP) with anti-BPA aptamer and trigger sequence was designed for BPA recognition and signal amplification. Next, under the action of BPA, the trigger probe was liberated from RP to initiate RCA reaction as primary amplification. Subsequently, the RCA products were used to trigger Exo III assisted secondary amplification with the help of hairpin probes, producing plenty of "G-quadruplex" in lantern-like structures. Finally, the continuously enriched "G-quadruplex lanterns" were lightened by zinc(II)-protoporphyrin IX (ZnPPIX) generating enhanced fluorescence signals. By integrating the primary RCA and secondary Exo III mediated cascade amplification strategy, this method displayed an excellent sensitivity with the detection limits of 5.4 × 10 -17 M. In addition, the anti-BPA aptamer exhibits high recognition ability with BPA, guaranteeing the specificity of detection. The reporter signal probe (G-quadruplex with ZnPPIX) provides a label-free fluorescence signals readout without complicated labeling procedures, making the method simple in design and cost-effective in operation. Moreover, environmental samples analysis was also performed, suggesting that our strategy was reliable and had a great potential application in environmental monitoring.

  19. A Label-Free and Sensitive Fluorescent Qualitative Assay for Bisphenol A Based on Rolling Circle Amplification/Exonuclease III-Combined Cascade Amplification

    Directory of Open Access Journals (Sweden)

    Xia Li

    2016-10-01

    Full Text Available Bisphenol A (BPA detection in drinking water and food packaging materials has attracted much attention since the discovery that BPA can interfere with normal physiological processes and cause adverse health effects. Here, we constructed a label-free aptamer fluorescent assay for selective and sensitive detection of BPA based on the rolling circle amplification (RCA/Exonuclease III (Exo III-combined cascade amplification strategy. First, the duplex DNA probe (RP with anti-BPA aptamer and trigger sequence was designed for BPA recognition and signal amplification. Next, under the action of BPA, the trigger probe was liberated from RP to initiate RCA reaction as primary amplification. Subsequently, the RCA products were used to trigger Exo III assisted secondary amplification with the help of hairpin probes, producing plenty of “G-quadruplex” in lantern-like structures. Finally, the continuously enriched “G-quadruplex lanterns” were lightened by zinc(II-protoporphyrin IX (ZnPPIX generating enhanced fluorescence signals. By integrating the primary RCA and secondary Exo III mediated cascade amplification strategy, this method displayed an excellent sensitivity with the detection limits of 5.4 × 10−17 M. In addition, the anti-BPA aptamer exhibits high recognition ability with BPA, guaranteeing the specificity of detection. The reporter signal probe (G-quadruplex with ZnPPIX provides a label-free fluorescence signals readout without complicated labeling procedures, making the method simple in design and cost-effective in operation. Moreover, environmental samples analysis was also performed, suggesting that our strategy was reliable and had a great potential application in environmental monitoring.

  20. Ultrafast Capillary Electrophoresis Isolation of DNA Aptamer for the PCR Amplification-Based Small Analyte Sensing

    Directory of Open Access Journals (Sweden)

    Emmanuelle eFiore

    2015-08-01

    Full Text Available Here, we report a new homogeneous DNA amplification-based aptamer assay for small analyte sensing. The aptamer of adenosine chosen as the model analyte was split into two fragments able to assemble in the presence of target. Primers were introduced at extremities of one fragment in order to generate the amplifiable DNA component. The amount of amplifiable fragment was quantifiable by Real-Time Polymerase Chain Reaction (RT-PCR amplification and directly reliable on adenosine concentration. This approach combines the very high separation efficiency and the homogeneous format (without immobilization of capillary electrophoresis and the sensitivity of real time PCR amplification. An ultrafast isolation of target-bound split aptamer (60 s was developed by designing a capillary electrophoresis input/ouput scheme. Such method was successfully applied to the determination of adenosine with a LOD of 1 µM.

  1. Single-Cell Whole-Genome Amplification and Sequencing: Methodology and Applications.

    Science.gov (United States)

    Huang, Lei; Ma, Fei; Chapman, Alec; Lu, Sijia; Xie, Xiaoliang Sunney

    2015-01-01

    We present a survey of single-cell whole-genome amplification (WGA) methods, including degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), multiple displacement amplification (MDA), and multiple annealing and looping-based amplification cycles (MALBAC). The key parameters to characterize the performance of these methods are defined, including genome coverage, uniformity, reproducibility, unmappable rates, chimera rates, allele dropout rates, false positive rates for calling single-nucleotide variations, and ability to call copy-number variations. Using these parameters, we compare five commercial WGA kits by performing deep sequencing of multiple single cells. We also discuss several major applications of single-cell genomics, including studies of whole-genome de novo mutation rates, the early evolution of cancer genomes, circulating tumor cells (CTCs), meiotic recombination of germ cells, preimplantation genetic diagnosis (PGD), and preimplantation genomic screening (PGS) for in vitro-fertilized embryos.

  2. Detection of genetically modified organisms (GMOs) using isothermal amplification of target DNA sequences.

    Science.gov (United States)

    Lee, David; La Mura, Maurizio; Allnutt, Theo R; Powell, Wayne

    2009-02-02

    The most common method of GMO detection is based upon the amplification of GMO-specific DNA amplicons using the polymerase chain reaction (PCR). Here we have applied the loop-mediated isothermal amplification (LAMP) method to amplify GMO-related DNA sequences, 'internal' commonly-used motifs for controlling transgene expression and event-specific (plant-transgene) junctions. We have tested the specificity and sensitivity of the technique for use in GMO studies. Results show that detection of 0.01% GMO in equivalent background DNA was possible and dilutions of template suggest that detection from single copies of the template may be possible using LAMP. This work shows that GMO detection can be carried out using LAMP for routine screening as well as for specific events detection. Moreover, the sensitivity and ability to amplify targets, even with a high background of DNA, here demonstrated, highlights the advantages of this isothermal amplification when applied for GMO detection.

  3. Absorption and distribution of calcium (labelled with 45Ca) applied after harvest to Asian pear (Pyrus pyrifolia) fruits

    International Nuclear Information System (INIS)

    Bhat, A.R.; Dhatt, A.S.; Dhillon, K.S.; Dhillon, S.K.

    1993-01-01

    Most of the disorders related to calcium are due to its unequal distribution within the plant rather than poor uptake. Immediately after fruit set, calcium uptake by roots as well as its translocation to developing fruit is extensive. However, the absorbed calcium does not reach the fruit in desired amounts during the subsequent fruit development stages and thereby, resulting in physiological disorders. Some corky spots underneath the peel of the fruit at harvest or during storage have been observed in Asian pear (cv. Patharnakh) which is extensively grown in northern India. There is need to increase the Ca concentration in the body of the fruit to prolong its life. The present investigation was, therefore, undertaken to study the absorption and distribution of post-harvest applied Ca in Asian pear fruit using radiotracer techniques. (author). 6 refs., 3 tabs

  4. chipPCR: an R package to pre-process raw data of amplification curves.

    Science.gov (United States)

    Rödiger, Stefan; Burdukiewicz, Michał; Schierack, Peter

    2015-09-01

    Both the quantitative real-time polymerase chain reaction (qPCR) and quantitative isothermal amplification (qIA) are standard methods for nucleic acid quantification. Numerous real-time read-out technologies have been developed. Despite the continuous interest in amplification-based techniques, there are only few tools for pre-processing of amplification data. However, a transparent tool for precise control of raw data is indispensable in several scenarios, for example, during the development of new instruments. chipPCR is an R: package for the pre-processing and quality analysis of raw data of amplification curves. The package takes advantage of R: 's S4 object model and offers an extensible environment. chipPCR contains tools for raw data exploration: normalization, baselining, imputation of missing values, a powerful wrapper for amplification curve smoothing and a function to detect the start and end of an amplification curve. The capabilities of the software are enhanced by the implementation of algorithms unavailable in R: , such as a 5-point stencil for derivative interpolation. Simulation tools, statistical tests, plots for data quality management, amplification efficiency/quantification cycle calculation, and datasets from qPCR and qIA experiments are part of the package. Core functionalities are integrated in GUIs (web-based and standalone shiny applications), thus streamlining analysis and report generation. http://cran.r-project.org/web/packages/chipPCR. Source code: https://github.com/michbur/chipPCR. stefan.roediger@b-tu.de Supplementary data are available at Bioinformatics online. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  5. Anthocyanin biosynthesis in pears is regulated by a R2R3-MYB transcription factor PyMYB10.

    Science.gov (United States)

    Feng, Shouqian; Wang, Yanling; Yang, Song; Xu, Yuting; Chen, Xuesen

    2010-06-01

    Skin color is an important factor in pear breeding programs. The degree of red coloration is determined by the content and composition of anthocyanins. In plants, many MYB transcriptional factors are involved in regulating anthocyanin biosynthesis. In this study, a R2R3-MYB transcription factor gene, PyMYB10, was isolated from Asian pear (Pyrus pyrifolia) cv. 'Aoguan'. Sequence analysis suggested that the PyMYB10 gene was an ortholog of MdMYB10 gene, which regulates anthocyanin biosynthesis in red fleshed apple (Malus x domestica) cv. 'Red Field'. PyMYB10 was identified at the genomic level and had three exons, with its upstream sequence containing core sequences of cis-acting regulatory elements involved in light responsiveness. Fruit bagging showed that light could induce expression of PyMYB10 and anthocyanin biosynthesis. Quantitative real-time PCR revealed that PyMYB10 was predominantly expressed in pear skins, buds, and young leaves, and the level of transcription in buds was higher than in skin and young leaves. In ripening fruits, the transcription of PyMYB10 in the skin was positively correlated with genes in the anthocyanin pathway and with anthocyanin biosynthesis. In addition, the transcription of PyMYB10 and genes of anthocyanin biosynthesis were more abundant in red-skinned pear cultivars compared to blushed cultivars. Transgenic Arabidopsis plants overexpressing PyMYB10 exhibited ectopic pigmentation in immature seeds. The study suggested that PyMYB10 plays a role in regulating anthocyanin biosynthesis and the overexpression of PyMYB10 was sufficient to induce anthocyanin accumulation.

  6. Rapid and nondestructive detection of watercore and sugar content in Asian pear by near infrared spectroscopy for commercial trade

    Directory of Open Access Journals (Sweden)

    Ronnarit Rittiron

    2014-11-01

    Full Text Available Watercore and sugar content are internal qualities which are impossible for exterior determination. Therefore the aims of this study were to develop models for nondestructive detection of watercore and predicting sugar content in pear using Near Infrared Spectroscopy (NIR technique. A total of 93 samples of Asian pear variety "SH-078" were used. For sugar content, spectrum of each fruit was measured in the short wavelength region (700–1100 nm in the reflection mode and the first derivative of spectra were then correlated with the sugar content in juice determined by digital refractometer. Prediction equation was performed by multiple linear regression. The result showed Standard Error of Prediction (SEP = 0.58°Bx, and Bias=0.11. The result from t-test showed that sugar content predicted by NIR was not significantly different from the value analyzed by refractometer at 95% confidence. For watercore disorder, NIR measurement was performed over the short wavelength range (700–850 nm in the transmission mode. The first derivative spectra were correlated with internal qualities. Then principle component analysis (PCA and partial least squares discriminant analysis (PLSDA were used to perform discrimination models. The accuracy of the PCA model was greater than the PLSDA one. The scores from PC1 were separated into two boundaries, one predicted rejected pears with 100% classification accuracy, and the other was accepted pears with 92% accuracy. The high accuracy of sugar content determining and watercore detecting by NIR reveal the high efficiency of NIR technique for detecting other internal qualities of fruit in the future.

  7. Adaptations of an avian supertramp: distribution, ecology, and life history of the pearly-eyed thrasher (Margarops fuscatus).

    Science.gov (United States)

    Wayne J. Arendt

    2006-01-01

    Part B The pearly-eyed thrasher is a major nest predator and competitor for nest sites of the endangered Puerto Rican Parrot. In all aspects of its distribution and ecology, Margarops fuscatus is a classic example of an avian “supertramp.” It is a pugnacious, highly vagile species, i.e., a good disperser, with a propensity to fill vacant or underexploited niches at all...

  8. Caught in the middle with multiple displacement amplification: the myth of pooling for avoiding multiple displacement amplification bias in a metagenome.

    Science.gov (United States)

    Marine, Rachel; McCarren, Coleen; Vorrasane, Vansay; Nasko, Dan; Crowgey, Erin; Polson, Shawn W; Wommack, K Eric

    2014-01-30

    Shotgun metagenomics has become an important tool for investigating the ecology of microorganisms. Underlying these investigations is the assumption that metagenome sequence data accurately estimates the census of microbial populations. Multiple displacement amplification (MDA) of microbial community DNA is often used in cases where it is difficult to obtain enough DNA for sequencing; however, MDA can result in amplification biases that may impact subsequent estimates of population census from metagenome data. Some have posited that pooling replicate MDA reactions negates these biases and restores the accuracy of population analyses. This assumption has not been empirically tested. Using mock viral communities, we examined the influence of pooling on population-scale analyses. In pooled and single reaction MDA treatments, sequence coverage of viral populations was highly variable and coverage patterns across viral genomes were nearly identical, indicating that initial priming biases were reproducible and that pooling did not alleviate biases. In contrast, control unamplified sequence libraries showed relatively even coverage across phage genomes. MDA should be avoided for metagenomic investigations that require quantitative estimates of microbial taxa and gene functional groups. While MDA is an indispensable technique in applications such as single-cell genomics, amplification biases cannot be overcome by combining replicate MDA reactions. Alternative library preparation techniques should be utilized for quantitative microbial ecology studies utilizing metagenomic sequencing approaches.

  9. Optical nano-biosensing interface via nucleic acid amplification strategy: construction and application.

    Science.gov (United States)

    Zhou, Hong; Liu, Jing; Xu, Jing-Juan; Zhang, Shu-Sheng; Chen, Hong-Yuan

    2018-03-21

    Modern optical detection technology plays a critical role in current clinical detection due to its high sensitivity and accuracy. However, higher requirements such as extremely high detection sensitivity have been put forward due to the clinical needs for the early finding and diagnosing of malignant tumors which are significant for tumor therapy. The technology of isothermal amplification with nucleic acids opens up avenues for meeting this requirement. Recent reports have shown that a nucleic acid amplification-assisted modern optical sensing interface has achieved satisfactory sensitivity and accuracy, high speed and specificity. Compared with isothermal amplification technology designed to work completely in a solution system, solid biosensing interfaces demonstrated better performances in stability and sensitivity due to their ease of separation from the reaction mixture and the better signal transduction on these optical nano-biosensing interfaces. Also the flexibility and designability during the construction of these nano-biosensing interfaces provided a promising research topic for the ultrasensitive detection of cancer diseases. In this review, we describe the construction of the burgeoning number of optical nano-biosensing interfaces assisted by a nucleic acid amplification strategy, and provide insightful views on: (1) approaches to the smart fabrication of an optical nano-biosensing interface, (2) biosensing mechanisms via the nucleic acid amplification method, (3) the newest strategies and future perspectives.

  10. NAIMA: target amplification strategy allowing quantitative on-chip detection of GMOs.

    Science.gov (United States)

    Morisset, Dany; Dobnik, David; Hamels, Sandrine; Zel, Jana; Gruden, Kristina

    2008-10-01

    We have developed a novel multiplex quantitative DNA-based target amplification method suitable for sensitive, specific and quantitative detection on microarray. This new method named NASBA Implemented Microarray Analysis (NAIMA) was applied to GMO detection in food and feed, but its application can be extended to all fields of biology requiring simultaneous detection of low copy number DNA targets. In a first step, the use of tailed primers allows the multiplex synthesis of template DNAs in a primer extension reaction. A second step of the procedure consists of transcription-based amplification using universal primers. The cRNA product is further on directly ligated to fluorescent dyes labelled 3DNA dendrimers allowing signal amplification and hybridized without further purification on an oligonucleotide probe-based microarray for multiplex detection. Two triplex systems have been applied to test maize samples containing several transgenic lines, and NAIMA has shown to be sensitive down to two target copies and to provide quantitative data on the transgenic contents in a range of 0.1-25%. Performances of NAIMA are comparable to singleplex quantitative real-time PCR. In addition, NAIMA amplification is faster since 20 min are sufficient to achieve full amplification.

  11. Optimization of edible coating formulations for improving postharvest quality and shelf life of pear fruit using response surface methodology.

    Science.gov (United States)

    Nandane, A S; Dave, Rudri K; Rao, T V Ramana

    2017-01-01

    The effect of composite edible films containing soy protein isolate (SPI) in combination with additives like hydroxypropyl methylcellulose (HPMC) and olive oil on 'Babughosha' pear ( Pyrus communis L.) stored at ambient temperature (28 ± 5 °C and 60 ± 10% RH) was evaluated using Response surface methodology (RSM). A total of 30 edible coating formulations comprising of SPI (2-6%, w/v), olive oil (0.7-1.1%, v/v), HPMC (0.1-0.5%, w/v) and potassium sorbate (0-0.4% w/v) were evaluated for optimizing the most suitable combination. Quality parameters like weight loss%, TSS, pH and titrable acidity of the stored pears were selected as response variables for optimization. The optimization procedure was carried out using RSM. It was observed that the response variables were mainly effected by concentration of SPI and olive oil in the formulation. Edible coating comprising of SPI 5%, HPMC 0.40%, olive oil 1% and potassium sorbate 0.22% was found to be most suitable combination for pear fruit with predicted values of response variables indicated as weight loss% 3.50, pH 3.41, TSS 11.13 and TA% 0.513.

  12. ESTIMATION OF AMPLIFICATION FACTOR IN EARTHQUAKE ENGINEERING

    Directory of Open Access Journals (Sweden)

    Nazarov Yuriy Pavlovich

    2015-03-01

    Full Text Available The authors are the developers of Odyssey Software (Eurosoft Co. for the analysis of seismological data and computing of seismic loads and their parameters. While communicating with the users of the software, the authors have revealed some uncertainty about both understanding of the term "amplification factor (AF" and calculation of the amplification factor using various methods. In this article, a simple example shows that the determination of the amplification factor as the ratio of the acceleration’s spectrum to the maximal acceleration is derived from the classical definition of AF in the form of the ratio of maximal dynamic displacement to the displacement by the action of static load. Deterministic and probabilistic ap-proaches for the calculating of the AF were discussed. There was an example of AFs calculation and their envelopes for translational and rotational components of seismic impact by using Odyssey Software.

  13. Amplification of hofmeister effect by alcohols.

    Science.gov (United States)

    Xu, Yun; Liu, Guangming

    2014-07-03

    We have demonstrated that Hofmeister effect can be amplified by adding alcohols to aqueous solutions. The lower critical solution temperature behavior of poly(N-isopropylacrylamide) has been employed as the model system to study the amplification of Hofmeister effect. The alcohols can more effectively amplify the Hofmeister effect following the series methanol alcohols and following the series d-sorbitol ≈ xylitol ≈ meso-erythritol alcohols. Our study reveals that the relative extent of amplification of Hofmeister effect is determined by the stability of the water/alcohol complex, which is strongly dependent on the chemical structure of alcohols. The more stable solvent complex formed via stronger hydrogen bonds can more effectively differentiate the anions through the anion-solvent complex interactions, resulting in a stronger amplification of Hofmeister effect. This study provides an alternative method to tune the relative strength of Hofmeister effect besides salt concentration.

  14. Lidar using the backscatter amplification effect

    Science.gov (United States)

    Razenkov, Igor A.; Banakh, Victor A.

    2018-04-01

    Experimental data proving the possibility of lidar measurement of the refractive turbulence strength based on the effect of backscatter amplification (BSA) are reported. It is shown that the values of the amplification factor correlate with the variance of random jitter of optical image of an incoherent light source depending on the value of the structure constant of the air refractive index turbulent fluctuations averaged over the probing path. This paper presents the results of measurements of the BSA factor in comparison with the simultaneous measurements of the BSA peak, which is very narrow and only occurs on the laser beam axis. It is constructed the range-time images of the derivative of the amplification factor gives a comprehensive picture of the location of turbulent zones and their temporal dynamics.

  15. Bio-cultural anchorage of the prickly pear cactus in Tlalnepantla (Morelos, Mexico

    Directory of Open Access Journals (Sweden)

    Torres-Salcido, Gerardo

    2016-06-01

    Full Text Available The prickly pear cactus is a source of food with strong bio-cultural anchorage in Mexico. This is due to at least three factors: 1 the nature and heritage of cacti; 2 cultural heritage; and 3 the socio-cultural relationships with historical and symbolic roots that have facilitated knowledge of how to cultivate it and how to use it. The aim of this article is to put factors of territorial anchorage and its historical transformation in context by examining the case of the municipality of Tlalnepantla in the state of Morelos, Mexico. This community has experienced accelerated change due to the exchange of traditional crops for the prickly pear cactus and the integration of farming, commercialization and agro-transformation. Our hypothesis is that the market, internal conflicts and a lack of socio-institutional coordination have put social organization into crisis, favoring the territorial spread of the prickly pear cactus and making the Local Agro-Food Systems (LAFS of Tlalnepantla less competitive. The conclusions highlight important economic and social advances whose roots lie in the strengthening and anchorage of the territory-product. However, circumstances both internal and external to the community persist, such as intra-community conflicts, the international market and cultural paradigm shifts that affect the producers and put consolidation of the LAFS at risk.El nopal es un alimento con un fuerte anclaje bio-cultural en México, propiciado por al menos tres factores: 1 la naturaleza y el patrimonio de cactáceas; 2 el patrimonio cultural; y, 3 las relaciones socio-culturales que han permitido un “saber hacer” y un “saber utilizar” con raíces históricas y simbólicas. El objetivo es situar los factores de anclaje territorial y su transformación histórica tomando como caso el municipio de Tlalnepantla, en el estado de Morelos, México. Esta comunidad ha experimentado un acelerado cambio por la reconversión de los cultivos

  16. Synergistic effects of some essential oils against fungal spoilage on pear fruit.

    Science.gov (United States)

    Nikkhah, Mehdi; Hashemi, Maryam; Habibi Najafi, Mohammad B; Farhoosh, Reza

    2017-09-18

    The development of natural protective agents as alternatives to chemical fungicides is currently in the spotlight. In the present investigation, chemical composition and antifungal activities of thyme, cinnamon, rosemary and marjoram essential oils (EO), as well as synergism of their possible double and triple combinations were investigated. The compositions of the oils were determined by GC/MS. For determination of antifungal activity against Penicillium expansum and Botrytis cinerea, a broth microdilution method was used. The possible interactions of some essential oil combinations were performed by the two and three-dimensional checkerboard assay and isobologram construction. An in vivo antifungal assay was performed by artificial wounding of pear fruits. The maximum antifungal activity was demonstrated by thyme and cinnamon oils which displayed lower MIC values whereas rosemary and marjoram oils with MIC range between 2500 and 10,000μg/mL exhibited weak antifungal activities against tested fungi. In synergy testing, some double combinations (thyme/cinnamon, thyme/rosemary, cinnamon/rosemary) were found to be synergistic (FICi≤0.5). The triple combination of thyme, cinnamon and rosemary was synergistic for B. cinerea and P. expansum (FICi values of 0.5 and 0.375, respectively); while combination of cinnamon, marjoram and thyme exhibited additive and synergistic effect against P. expansum (FIC=0.625) and B. cinerea (FIC=0.375) respectively. The usage of a mathematical Gompertz model in relation to fungal kinetics, showed that the model could be used to predict growth curves (R 2 =0.993±0.05). For B. cinerea, Gompertz parameters for double and triple combination treatments showed significant increase in lag phase (1.92 and 2.92days, respectively) compared to single treatments. Increase lag time up to 2.82days (P<0.05) also observed in P. expansum treated by triple combination of EOs. Base on the results, the lowest maximum growth rate (0.37mm/day) was observed

  17. [Colorimetric detection of HPV6 and HPV16 by loop mediated isothermal amplification].

    Science.gov (United States)

    Lu, Chun-bin; Luo, Le; Yang, Meng-jie; Nie, Kai; Wang, Miao; Ma, Xue-Jun

    2011-01-01

    A simple, rapid and sensitive colorimetric loop mediated isothermal amplification (LAMP) method was established to detect HPV6 and HPV 16 respectively. The method employed a set of four specially designed primers that recognized six distinct sequences of HPV6-E6 or HPV16-E7 for amplification of nucleic acid under isothermal conditions at 63 degrees C for one hour. The amplification process of LAMP was monitored by the addition of HNB (hydroxy naphthol blue) dye prior to amplification. A positive reaction was indicated by a color change from violet to sky blue and confirmed by real-time turbidimeter and agarose electrophoresis. Thirteen cervical swab samples having single infection with 13 different HPV genotypes were examined to evaluate the specificity. A serial dilution of a cloned plasmid containing HPV-E6 or HPV-E7 gene was examined to evaluate the sensitivity. The results showed that no cross-reaction with other HPV genotypes was observed. The colorimetric LAMP assay could achieve a sensitivity of 1000 copies, 10-20 times lower than that of real-time PCR. The assay was further evaluated with 62 clinical specimens and consistent results were obtained compared with the detection using Kai Pu HPV Genotyping Kit. We concluded that this colorimetric LAMP assay had potential usefulness for the rapid screening of the HPV6 or HPV16 infection in the laboratories and hospitals of provincial and municipal region in China.

  18. Nanoliter reactors improve multiple displacement amplification of genomes from single cells.

    Directory of Open Access Journals (Sweden)

    Yann Marcy

    2007-09-01

    Full Text Available Since only a small fraction of environmental bacteria are amenable to laboratory culture, there is great interest in genomic sequencing directly from single cells. Sufficient DNA for sequencing can be obtained from one cell by the Multiple Displacement Amplification (MDA method, thereby eliminating the need to develop culture methods. Here we used a microfluidic device to isolate individual Escherichia coli and amplify genomic DNA by MDA in 60-nl reactions. Our results confirm a report that reduced MDA reaction volume lowers nonspecific synthesis that can result from contaminant DNA templates and unfavourable interaction between primers. The quality of the genome amplification was assessed by qPCR and compared favourably to single-cell amplifications performed in standard 50-microl volumes. Amplification bias was greatly reduced in nanoliter volumes, thereby providing a more even representation of all sequences. Single-cell amplicons from both microliter and nanoliter volumes provided high-quality sequence data by high-throughput pyrosequencing, thereby demonstrating a straightforward route to sequencing genomes from single cells.

  19. Rapid detection of Mycobacterium tuberculosis by recombinase polymerase amplification.

    Directory of Open Access Journals (Sweden)

    David S Boyle

    Full Text Available Improved access to effective tests for diagnosing tuberculosis (TB has been designated a public health priority by the World Health Organisation. In high burden TB countries nucleic acid based TB tests have been restricted to centralised laboratories and specialised research settings. Requirements such as a constant electrical supply, air conditioning and skilled, computer literate operators prevent implementation of such tests in many settings. Isothermal DNA amplification technologies permit the use of simpler, less energy intensive detection platforms more suited to low resource settings that allow the accurate diagnosis of a disease within a short timeframe. Recombinase Polymerase Amplification (RPA is a rapid, low temperature isothermal DNA amplification reaction. We report here RPA-based detection of Mycobacterium tuberculosis complex (MTC DNA in <20 minutes at 39 °C. Assays for two MTC specific targets were investigated, IS6110 and IS1081. When testing purified MTC genomic DNA, limits of detection of 6.25 fg (IS6110 and 20 fg (IS1081were consistently achieved. When testing a convenience sample of pulmonary specimens from suspected TB patients, RPA demonstrated superior accuracy to indirect fluorescence microscopy. Compared to culture, sensitivities for the IS1081 RPA and microscopy were 91.4% (95%CI: 85, 97.9 and 86.1% (95%CI: 78.1, 94.1 respectively (n = 71. Specificities were 100% and 88.6% (95% CI: 80.8, 96.1 respectively. For the IS6110 RPA and microscopy sensitivities of 87.5% (95%CI: 81.7, 93.2 and 70.8% (95%CI: 62.9, 78.7 were obtained (n = 90. Specificities were 95.4 (95% CI: 92.3,98.1 and 88% (95% CI: 83.6, 92.4 respectively. The superior specificity of RPA for detecting tuberculosis was due to the reduced ability of fluorescence microscopy to distinguish Mtb complex from other acid fast bacteria. The rapid nature of the RPA assay and its low energy requirement compared to other amplification technologies suggest RPA-based TB

  20. Shelf-life extension of pear (Pyrus communis L.) by gamma-irradiation

    International Nuclear Information System (INIS)

    Wani, A.M.; Hussain, P.R.; Dar, M.A.; Mir, M.A.

    2007-01-01

    Hard and unripe pear fruit was precooled and subjected to γ-irradiation in the range of 0.1-2.5 kGy. After irradiation fruit was stored under ambient conditions (25±2 degC, RH 70%) and evaluated periodically for firmness, total soluble solids, titratable acidity, chlorophyll, physiological loss in weight, visual colour score, overall acceptability and microbial load. γ-irradiation at 0.1-0.9 kGy did not show any significant effect with respect to shelf-life extension and microbial load. However, irradiation doses of 1.0-1.5 kGy significantly (p≤0.05) delayed ripening and did not spoil for 14 days of storage under ambient conditions. The microbial load was reduced without affecting the physico-chemical and sensory attributes. The irradiation doses of 2.0-2.5 kGy resulted in reduced microbial load and delayed ripening but adversely affected the fruit colour. (author)

  1. Nitrogen replenishment using variable rate application technique in a small hand-harvested pear orchard

    International Nuclear Information System (INIS)

    Vatsanidou, A.; Nanos, G.D.; Fountas, S.; Baras, J.; Castrignano, A.; Gemtos, T.A.

    2017-01-01

    Precision agriculture is a management approach for sustainable agriculture. It can be applied even in small fields. It aims to optimize inputs, improve profits, and reduce adverse environmental impacts. In this study, a series of measurements were conducted over three growing seasons to assess variability in a 0.55 ha pear orchard located in central Greece. Soil ECa was measured using EM38 sensor, while soil samples were taken from a grid 17 × 8 m and analysed for texture, pH, P, K, Mg, CaCO3, and organic matter content. Data analysis indicated that most of the nutrients were at sufficient levels. Soil and yield maps showed considerable variability while fruit quality presented small variations across the orchard. Yield fluctuations were observed, possibly due to climatic conditions. Prescription maps were developed for nitrogen variable rate application (VRA) for two years based on the replacement of the nutrients removed by the crop. VRA application resulted in 56% and 50% reduction of N fertiliser compared to uniform application.

  2. Nitrogen replenishment using variable rate application technique in a small hand-harvested pear orchard

    Energy Technology Data Exchange (ETDEWEB)

    Vatsanidou, A.; Nanos, G.D.; Fountas, S.; Baras, J.; Castrignano, A.; Gemtos, T.A.

    2017-07-01

    Precision agriculture is a management approach for sustainable agriculture. It can be applied even in small fields. It aims to optimize inputs, improve profits, and reduce adverse environmental impacts. In this study, a series of measurements were conducted over three growing seasons to assess variability in a 0.55 ha pear orchard located in central Greece. Soil ECa was measured using EM38 sensor, while soil samples were taken from a grid 17 × 8 m and analysed for texture, pH, P, K, Mg, CaCO3, and organic matter content. Data analysis indicated that most of the nutrients were at sufficient levels. Soil and yield maps showed considerable variability while fruit quality presented small variations across the orchard. Yield fluctuations were observed, possibly due to climatic conditions. Prescription maps were developed for nitrogen variable rate application (VRA) for two years based on the replacement of the nutrients removed by the crop. VRA application resulted in 56% and 50% reduction of N fertiliser compared to uniform application.

  3. EFFECTS OF PLANTING DENSITYAND ORGANIC FERTILIZATION DOSES ON PRODUCTIVE EFFICIENCY OF CACTUS PEAR

    Directory of Open Access Journals (Sweden)

    NALÍGIA GOMES DE MIRANDA E SILVA

    2016-01-01

    Full Text Available Cactus is crucial for the livestock of semi - arid regions in Brazil. This plant has shown the high productivity of forage, which is influenced by several management factors. This study aimed to evaluate the effect of organic fertilization doses (20, 40 and 80 t/ ha of bovine manure/ha/two years and planting densities (20, 40, 80 and 160 thousand plants/ha on the productivity of cactus pear Clone IPA - 20 ( Opuntia ficus - indica Mill. At the Experimental Station of Caruaru at the Agronomic Institute of Pernambuco, IPA has conducted the experiment. The experimental design was randomized blocks, with split plot arrangements. Higher shoot productivity was observed with increased population density and the application of manure at 80 t ha - 1two years - 1 with values of 61, 90, 117 and 139 t DM ha - 1 two years - 1 at densities of 20, 40, 80 and 160,000 plants ha - 1. The planting density influenced the productivity of cladode - plant and root dry weight, showing exponential responses, with higher cladode - plant and roots weight by area observed with increased plant density. The efficiency of organic fertilization decreased with the increase in manure doses. For increase cactus productivity, 40 t of bovine manure ha - 1 two years - 1 for plantations with 160,000 plants/ha is recommended.

  4. Nutritive value and chemical composition of prickly pear seeds (Opuntia ficus indica L.) growing in Turkey.

    Science.gov (United States)

    Özcan, Mehmet Musa; Al Juhaimi, Fahad Y

    2011-08-01

    The proximate composition and physico-chemical properties (moisture, crude lipid, crude protein, ash, and crude fiber, peroxide value, saponification value, acidity, relative density and refractive index) of prickly pear seed and corresponding oil were determined. The mineral contents (Ca, Cu, Fe, K, Mg, Na, P, Mn and Zn) of samples were analyzed by inductively coupled plasma atomic emission spectrometry. Minerals determined were: calcium 471.2 mg/kg, potassium 532.7 mg/kg, magnesium 117.3 mg/kg, phosphorus 1,627.5 mg/kg and natrium 71.3 mg/kg. The fatty acid profiles of seed oil from the Opuntia ficus indica were analyzed by gas chromatography. Linoleic acid was established as the major fatty acid (61.01%), followed by oleic (25.52%) and palmitic (12.23%) acids. Both myristic, stearic and arachidonic acids were detected in O. ficus indica seed oil in low amounts. As a result, O. ficus indica seeds are an important source of natural fiber and, given its high linoleic acid content, its oil can be used as a nutraceutic agent.

  5. Micromorphology of cactus-pear (Opuntia ficus-indica (L.) Mill) cladodes based on scanning microscopies.

    Science.gov (United States)

    Ben Salem-Fnayou, Asma; Zemni, Hassène; Nefzaoui, Ali; Ghorbel, Abdelwahed

    2014-01-01

    Cladode ultrastructural features of two prickly and two spineless Opuntia ficus-indica cultivars were examined using environmental scanning electron and atomic force microscopies. Observations focused on cladode as well as spine and glochid surface micromorphologies. Prickly cultivars were characterized by abundant cracked epicuticular wax deposits covering the cladode surface, with an amorphous structure as observed by AFM, while less abundant waxy plates were observed by ESEM on spineless cultivar cladodes. Further AFM observations allowed a rough granular and crystalloid epicuticular wax structure to be distinguished in spineless cultivars. Regarding spine micromorphology, prickly cultivars had strong persistent spines, observed by ESEM as a compact arrangement of oblong epidermal cells with a rough granular structure. However, deciduous spines in spineless cultivars had a broken transversely fissured epidermis covering a parallel arrangement of fibres. Through AFM, the deciduous spine surface presented an irregular hilly and smooth microrelief while persistent spines exhibited rough helical filamentous prints. ESEM and AFM studies of cladode surfaces from prickly and spineless cactus pear cultivars revealed valuable micro-morphological details that ought to be extended to a large number of O. ficus-indica cultivars. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Effects of the pear tree canopy on photosynthetically active radiation availability

    International Nuclear Information System (INIS)

    Rossi, F.; Baldini, E.; Baraldi, R.

    1984-01-01

    The relationships existing between radiant energy and photosynthesis have been extensively investigated on the apple /2/ but not on the other fruit trees, pear included. In addition, such information resists generalization, owing to the remarkable differences underlying tree morphology and physiology of the different species; furthermore, some disagreement arises regarding the terminology and the units used to evaluate the amount of radiant energy useful for the photosynthetic process. In general this evaluation is based on the readouts of illuminance (symbol Ev; unit: lux), in agreement with the photopic curve (fig. 1:A), i.e. with the human eye sensibility to the visible radiation(light). However, the relative response of the chloroplasts to the radiant flux, although included within the same spectral wavebands as the photopic curve, follows a different model (fig.1:B), that is, it has two peaks in connection with the spectral wavelenghts of blue (440–490 nm), and, particularly, of red (620–700 nm). Therefore, according to a number of authors /3/6/8/11/, the correct evaluation of the photosynthetically active radiation should be made using sensors calibrated to measure the photosynthetic photon lux density (symbol: PPFD; unit: μE m -2 s -1 ), and provided with a relative spectral response similar to that of the leaves. (author)

  7. Antibacterial and antioxidant activities in extracts of fully grown cladodes of 8 cultivars of cactus pear.

    Science.gov (United States)

    Sánchez, E; Dávila-Aviña, J; Castillo, S L; Heredia, N; Vázquez-Alvarado, R; García, S

    2014-04-01

    The antimicrobial and antioxidant activities of some cultivars of the nopal cactus have not been determined. In this study, 8 cultivars of nopal cacti from Mexico were assayed for phenolic content, antioxidant activities, and antimicrobial activities against Campylobacter Jejuni, Vibrio cholera, and Clostridium Perfringens. Plant material was washed, dried, and macerated in methanol. Minimum bactericidal concentrations (MBCs) were determined using the broth microdilution method. Antioxidant activities were quantitatively determined using spectrophotometric methods. The MCBs of the nopal cacti ranged from 1.1 to 12.5 mg/mL for c. jejuni, 4.4 to 30 mg/mL for V. cholera, and 0.8 to 16 mg/mL for C. perfringens in the cultivars Cardon Blanco, Real de Catorce, and Jalpa, respectively. High quantities of total phenols and total flavonoids were found in the Jalpa cacti (3.80 mg of gallic acid equivalent GAE/g dry weight [DW] and 36.64 mg of quercetin equivalents [QE]/g DW, respectively). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities (RSA) were correlated to bioactive compound contents. The Villanueva cacti had the highest %RSA at 42.31%, and the lowest activity was recorded in Copena V1 at 19.98%. In conclusion, we found that some of the 8 cactus pear cultivars studied may be used for their antioxidant compounds or antimicrobials to control or prevent the contamination of foods. © 2014 Institute of Food Technologists®

  8. Evaluation of selected nursery traits in combination rootstocks and variety for Asian pear trees

    Directory of Open Access Journals (Sweden)

    Tomáš Nečas

    2012-01-01

    Full Text Available Evaluation of the nursery traits of rootstocks and varieties is an important tool for optimisation of the fruit orchard plants final growing. The present experiment evaluated the standard and progressive rootstocks for the pear trees in combination with the Asian varieties of P. pyrifolia Nakai. a P. ussuriensis Maxim. in comparison with the control variety ‘Conference’ P. communis L. The evaluation was focused on such properties as affinity, growth of the tree propagated by budding, thickness of the root crown at the rootstock and the root suckering. Their interactions were tested statistically. The highest values of affinity within the nursery were achieved with the following rootstocks: ‘BET’ (96.6%, ‘FOX 11’ (95.5 %, and ‘MA’ (94.4 %. ‘Hosui’ was identified as the variety with the greatest growing capacity, with an average gain of 1238.2 mm. The most supported variety growth was exhibited by the ‘FOX 11’ rootstock, with an average gain of the trees propagated by budding of 1388.8 mm. The growth intensity itself was demonstrated most in the thickness of the root crown of the ‘BET’ (P. betulifolia rootstock, namely 15.8 mm. The rootstock that exhibited the strongest root suckering was also ‘BET’ with 6.2 suckers per rootstock. The identified differences between the varieties and rootstocks were confirmed as statistically significant or even highly statistically significant.

  9. Chemical And Physiological Studies On Drought Stress Tolerance Of Irradiated Communis Pear Using Tissue Culture

    International Nuclear Information System (INIS)

    Zaied, N.S.; Ragab, E.A.

    2007-01-01

    The rooted in vitro irradiated pear rootstocks (Pyrus communis) were subjected to drought stress by using different concentrations of mannitol (20, 40, 60, 80 and 100 gm/l), polyethylene glycol (PEG) at concentrations 2, 4, 6, 8 and 10 % to culture medium and also agar at concentrations 6, 8, 10, 12 and 14 gm/l to study their effects on tissue culture and chemical analysis and their tolerance to drought stress. The obtained results showed that the number of shoots, shoot length and number of leaves were higher at 20 and 40 gm/l mannitol. Increasing mannitol concentration enhanced the increase of chlorophyll b, reducing sugars, total indoles and total phenols up to the highest level at 100 gm/l. Adding PEG at concentration 2% to the culture medium encouraged significant increases in the number of shoots and number of leaves and increase chlorophyll a, and non-reducing sugars as well as significant decrease in number of shoots, shoots length, number of leaves, root length and number of roots with increasing agar concentrations to the culture medium. However, decreasing agar concentration in the culture medium induced increase in chlorophyll A and non-reducing sugar

  10. Agrobiodiversity of cactus pear (Opuntia, Cactaceae in the Meridional Highlands Plateau of Mexico

    Directory of Open Access Journals (Sweden)

    Juan Antonio Reyes-Agüero

    2011-08-01

    Full Text Available Mexico is characterized by a remarkable richness of Opuntia, mostly at the Meridional Highlands Plateau; it is also here where the greatest richness of Opuntia variants occurs. Most of these variants have been maintained in homegardens; however, the gathering process which originated these homegardens has been disrupted over the past decades, as a result of social change and the destruction of large wild nopaleras. If the variants still surviving in homegardens are lost, these will be hard to recover, that is, the millenary cultural heritage from the human groups that populated the Mexican Meridional Highland Plateau will be lost forever. This situation motivated the preparation of a catalogue that records the diversity of wild and cultivated Opuntia variants living in the meridional Highlands Plateau. To this end, 379 samples were obtained in 29 localities, between 1998 and 2003. The information was processed through Twinspan. All specimens were identified and preserved in herbaria. Botanical keys and descriptions were elaborated. The catalogue includes information on 126 variants comprising 18 species. There were species with only one variant (Opuntia atropes, O. cochinera, O. jaliscana, O. leucotricha, O. rzedowskii and O. velutina, two (O. durangensis, O. lindheimeri, O. phaeacantha and O. robusta, five (O. joconostle and O. lasiacantha, seven (O. chavena, 12 (O. hyptiacantha and O. streptacantha, 15 (O. ficus-indica, 22 (O. albicarpa, and up to 34 (O. megacantha. Additionally, 267 common cactus pear names were related to those variants.

  11. Nitrogen replenishment using variable rate application technique in a small hand-harvested pear orchard

    Directory of Open Access Journals (Sweden)

    Anna Vatsanidou

    2018-02-01

    Full Text Available Precision agriculture is a management approach for sustainable agriculture. It can be applied even in small fields. It aims to optimize inputs, improve profits, and reduce adverse environmental impacts. In this study, a series of measurements were conducted over three growing seasons to assess variability in a 0.55 ha pear orchard located in central Greece. Soil ECa was measured using EM38 sensor, while soil samples were taken from a grid 17 × 8 m and analysed for texture, pH, P, K, Mg, CaCO3, and organic matter content. Data analysis indicated that most of the nutrients were at sufficient levels. Soil and yield maps showed considerable variability while fruit quality presented small variations across the orchard. Yield fluctuations were observed, possibly due to climatic conditions. Prescription maps were developed for nitrogen variable rate application (VRA for two years based on the replacement of the nutrients removed by the crop. VRA application resulted in 56% and 50% reduction of N fertiliser compared to uniform application.

  12. Pectin isolated from prickly pear (Opuntia SSP) modifies LDL metabolism in cholesterol-fed guinea pigs

    International Nuclear Information System (INIS)

    Fernandez, M.L.; McNamara, D.J.

    1990-01-01

    The effects of dietary pectin on plasma and hepatic cholesterol (CH) levels, plasma lipoprotein profiles, hepatic 3-hydroxy-3-methylglutaryl Coenzyme A (HMG-CoA) reductase activity, and low density lipoprotein (LDL) binding to hepatic membranes were investigated by feeding 1% pectin to guinea pigs on a high CH diet. Animals were fed either chow + 0.25% CH (HC diet) or the CH diet + 1% prickly pear pectin (HC-P diet) for 25 days. Plasma CH levels were decreased 26% by the HC-P with 33% decreases in LDL and KDL. LDL peak density shifted from 1.040 to 1.055 g/ml with pectin. Hepatic total, free and esterified CH levels were reduced 60, 40 and 85% respectively by the HC-P diet. In contrast, HMG-CoA reductase activity was unaffected. 125 I-LDL binding to hepatic membranes was increased by intake of the HC-P diet compared to the HC diet. The affinity of the apo B/E receptor for LDL was not affected by dietary pectin while the receptor number was increased 1.5-fold in animals on the HC-P diet. These data suggest that the parameters of HC metabolism affected by dietary pectin are consistent with an increased demand on the hepatic CH pools which possibly results from increased fecal excretion of bile acids

  13. Amplification in Technical Manuals: Theory and Practice.

    Science.gov (United States)

    Killingsworth, M. Jimmie; And Others

    1989-01-01

    Examines how amplification (rhetorical techniques by which discourse is extended to enhance its appeal and information value) tends to increase and improve the coverage, rationale, warnings, behavioral alternatives, examples, previews, and general emphasis of technical manuals. Shows how classical and modern rhetorical theories can be applied to…

  14. Intelligence amplification framework for enhancing scheduling processes

    NARCIS (Netherlands)

    Dobrkovic, Andrej; Liu, Luyao; Iacob, Maria Eugenia; van Hillegersberg, Jos

    2016-01-01

    The scheduling process in a typical business environment consists of predominantly repetitive tasks that have to be completed in limited time and often containing some form of uncertainty. The intelligence amplification is a symbiotic relationship between a human and an intelligent agent. This

  15. Social amplification of risk: a conceptual framework

    International Nuclear Information System (INIS)

    Kasperson, R.E.; Renn, O.; Slovic, P.; Brown, H.S.; Emel, J.; Goble, R.; Kasperson, J.X.; Ratick, S.

    1988-01-01

    One of the most perplexing problems in risk analysis is why some relatively minor risks or risk events, as assessed by technical experts, often elicit strong public concerns and result in substantial impacts upon society and economy. This article sets forth a conceptual framework that seeks to link systematically the technical assessment of risk with psychological, sociological, and cultural perspectives of risk perception and risk-related behavior. The main thesis is that hazards interact with psychological, social, institutional, and cultural processes in ways that may amplify or attenuate public responses to the risk or risk event. A structural description of the social amplification of risk is now possible. Amplification occurs at two stages: in the transfer of information about the risk, and in the response mechanisms of society. Signals about risk are processed by individual and social amplification stations, including the scientist who communicates the risk assessment, the news media, cultural groups, interpersonal networks, and others. Key steps of amplifications can be identified at each stage. The amplified risk leads to behavioral responses, which, in turn, result in secondary impacts. Models are presented that portray the elements and linkages in the proposed conceptual framework

  16. Influence of rootstocks on growth, yield, fruit quality and leaf mineral element contents of pear cv. 'Santa Maria' in semi-arid conditions

    Directory of Open Access Journals (Sweden)

    Ali Ikinci

    2014-01-01

    Full Text Available BACKGROUND: Rootstocks play an essential role to determining orchard performance of fruit trees. Pyrus communisand Cydonia oblonga are widely used rootstocks for European pear cultivars. The lack of rootstocks adapted to different soil conditions and different grafted cultivars is widely acknowledged in pear culture. Cydonia rootstocks (clonal and Pyrus rootstocks (seedling or clonal have their advantages and disadvantages. In each case, site-specific environmental characteristics, specific cultivar response and production objectives must be considered before choosing the best rootstock. In this study, the influence of three Quince (BA 29, Quince A = MA, Quince C = MC and a local European pear seedling rootstocks on the scion yield, some fruit quality characteristics and leaf macro (N, P, K, Ca and Mg and micro element (Fe, Zn, Cu, Mn and B content of 'Santa Maria' pear (Pyrus communis L. were investigated. RESULTS: Trees on seedling rootstock had the highest annual yield, highest cumulative yield (kg tree−1, largest trunk cross-sectional area (TCSA, lowest yield efficiency and lowest cumulative yield (ton ha−1 in the 10th year after planting. The rootstocks had no significant effect on average fruit weight and fruit volume. Significantly higher fruit firmness was obtained on BA 29 and Quince A. The effect of rootstocks on the mineral element accumulation (N, K, Ca, Mg, Fe, Zn, Cu, Mn and B was significant. Leaf analysis showed that rootstocks used had different mineral uptake efficiencies throughout the early season. CONCLUSION: The results showed that the rootstocks strongly affected fruit yield, fruit quality and leaf mineral element uptake of 'Santa Maria' pear cultivar. Pear seedling and BA 29 rootstock found to be more prominent in terms of several characteristics for 'Santa Maria' pear cultivar that is grown in highly calcareous soil in semi-arid climate conditions. We determined the highest N, P (although insignificant, K, Ca, Mg, Fe

  17. Characterization of the lipoxygenase (LOX) gene family in the Chinese white pear (Pyrus bretschneideri) and comparison with other members of the Rosaceae.

    Science.gov (United States)

    Li, Meng; Li, Leiting; Dunwell, Jim M; Qiao, Xin; Liu, Xing; Zhang, Shaoling

    2014-06-07

    Lipoxygenases (LOXs), a type of non-haem iron-containing dioxygenase, are ubiquitous enzymes in plants and participate in the formation of fruit aroma which is a very important aspect of fruit quality. Amongst the various aroma volatiles, saturated and unsaturated alcohols and aldehydes provide the characteristic aroma of the fruit. These compounds are formed from unsaturated fatty acids through oxidation, pyrolysis and reduction steps. This biosynthetic pathway involves at least four enzymes, including LOX, the enzyme responsible for lipid oxidation. Although some studies have been conducted on the LOX gene family in several species including Arabidopsis, soybean, cucumber and apple, there is no information from pear; and the evolutionary history of this gene family in the Rosaceae is still not resolved. In this study we identified 107 LOX homologous genes from five Rosaceous species (Pyrus bretschneideri, Malus × domestica, Fragaria vesca, Prunus mume and Prunus persica); 23 of these sequences were from pear. By using structure analysis, phylogenic analysis and collinearity analysis, we identified variation in gene structure and revealed the phylogenetic evolutionary relationship of this gene family. Expression of certain pear LOX genes during fruit development was verified by analysis of transcriptome data. 23 LOX genes were identified in pear and these genes were found to have undergone a duplication 30-45 MYA; most of these 23 genes are functional. Specific gene duplication was found on chromosome4 in the pear genome. Useful information was provided for future research on the evolutionary history and transgenic research on LOX genes.

  18. Genomic characterization, phylogenetic comparison and differential expression of the cyclic nucleotide-gated channels gene family in pear (Pyrus bretchneideri Rehd.).

    Science.gov (United States)

    Chen, Jianqing; Yin, Hao; Gu, Jinping; Li, Leiting; Liu, Zhe; Jiang, Xueting; Zhou, Hongsheng; Wei, Shuwei; Zhang, Shaoling; Wu, Juyou

    2015-01-01

    The cyclic nucleotide-gated channel (CNGC) family is involved in the uptake of various cations, such as Ca(2+), to regulate plant growth and respond to biotic and abiotic stresses. However, there is far less information about this family in woody plants such as pear. Here, we provided a genome-wide identification and analysis of the CNGC gene family in pear. Phylogenetic analysis showed that the 21 pear CNGC genes could be divided into five groups (I, II, III, IVA and IVB). The majority of gene duplications in pear appeared to have been caused by segmental duplication and occurred 32.94-39.14 million years ago. Evolutionary analysis showed that positive selection had driven the evolution of pear CNGCs. Motif analyses showed that Group I CNGCs generally contained 26 motifs, which was the greatest number of motifs in all CNGC groups. Among these, eight motifs were shared by each group, suggesting that these domains play a conservative role in CNGC activity. Tissue-specific expression analysis indicated that functional diversification of the duplicated CNGC genes was a major feature of long-term evolution. Our results also suggested that the P-S6 and PBC & hinge domains had co-evolved during the evolution. These results provide valuable information to increase our understanding of the function, evolution and expression analyses of the CNGC gene family in higher plants. Copyright © 2014 Elsevier Inc. All rights reserved.

  19. Hydrogen sulfide prolongs postharvest storage of fresh-cut pears (Pyrus pyrifolia by alleviation of oxidative damage and inhibition of fungal growth.

    Directory of Open Access Journals (Sweden)

    Kang-Di Hu

    Full Text Available Hydrogen sulfide (H2S has proved to be a multifunctional signaling molecule in plants and animals. Here, we investigated the role of H2S in the decay of fresh-cut pears (Pyrus pyrifolia. H2S gas released by sodium hydrosulfide (NaHS prolonged the shelf life of fresh-cut pear slices in a dose-dependent manner. Moreover, H2S maintained higher levels of reducing sugar and soluble protein in pear slices. H2S significantly reduced the accumulation of hydrogen peroxide (H2O2, superoxide radicals (•O2(- and malondialdehyde (MDA. Further investigation showed that H2S fumigation up-regulated the activities of antioxidant enzymes ascorbate peroxidase (APX, catalase (CAT, and guaiacol peroxidase (POD, while it down-regulated those of lipoxygenase (LOX, phenylalanine ammonia lyase (PAL and polyphenol oxidase (PPO. Furthermore, H2S fumigation effectively inhibited the growth of two fungal pathogens of pear, Aspergillus niger and Penicillium expansum, suggesting that H2S can be developed as an effective fungicide for postharvest storage. The present study implies that H2S is involved in prolonging postharvest storage of pears by acting as an antioxidant and fungicide.

  20. Current Developments in Prokaryotic Single Cell Whole Genome Amplification

    Energy Technology Data Exchange (ETDEWEB)

    Goudeau, Danielle; Nath, Nandita; Ciobanu, Doina; Cheng, Jan-Fang; Malmstrom, Rex

    2014-03-14

    Our approach to prokaryotic single-cell Whole Genome Amplification at the JGI continues to evolve. To increase both the quality and number of single-cell genomes produced, we explore all aspects of the process from cell sorting to sequencing. For example, we now utilize specialized reagents, acoustic liquid handling, and reduced reaction volumes eliminate non-target DNA contamination in WGA reactions. More specifically, we use a cleaner commercial WGA kit from Qiagen that employs a UV decontamination procedure initially developed at the JGI, and we use the Labcyte Echo for tip-less liquid transfer to set up 2uL reactions. Acoustic liquid handling also dramatically reduces reagent costs. In addition, we are exploring new cell lysis methods including treatment with Proteinase K, lysozyme, and other detergents, in order to complement standard alkaline lysis and allow for more efficient disruption of a wider range of cells. Incomplete lysis represents a major hurdle for WGA on some environmental samples, especially rhizosphere, peatland, and other soils. Finding effective lysis strategies that are also compatible with WGA is challenging, and we are currently assessing the impact of various strategies on genome recovery.

  1. Recombinase polymerase amplification: Emergence as a critical molecular technology for rapid, low-resource diagnostics.

    Science.gov (United States)

    James, Ameh; Macdonald, Joanne

    2015-01-01

    Isothermal molecular diagnostics are bridging the technology gap between traditional diagnostics and polymerase chain reaction-based methods. These new techniques enable timely and accurate testing, especially in settings where there is a lack of infrastructure to support polymerase chain reaction facilities. Despite this, there is a significant lack of uptake of these technologies in developing countries where they are highly needed. Among these novel isothermal technologies, recombinase polymerase amplification (RPA) holds particular potential for use in developing countries. This rapid nucleic acid amplification approach is fast, highly sensitive and specific, and amenable to countries with a high burden of infectious diseases. Implementation of RPA technology in developing countries is critically required to assess limitations and potentials of the diagnosis of infectious disease, and may help identify impediments that prevent adoption of new molecular technologies in low resource- and low skill settings. This review focuses on approaching diagnosis of infectious disease with RPA.

  2. Amplification and overexpression of aurora kinase A (AURKA) in immortalized human ovarian epithelial (HOSE) cells.

    Science.gov (United States)

    Chung, C M; Man, C; Jin, Y; Jin, C; Guan, X Y; Wang, Q; Wan, T S K; Cheung, A L M; Tsao, S W

    2005-07-01

    Immortalization is an early and essential step of human carcinogenesis. Amplification of chromosome 20q has been shown to be a common event in immortalized cells and cancers. We have previously reported that gain and amplification of chromosome 20q is a non-random and common event in immortalized human ovarian surface epithelial (HOSE) cells. The chromosome 20q harbors genes including TGIF2 (20q11.2-q12), AIB1 (20q12), PTPN1 (20q13.1), ZNF217 (20q13.2), and AURKA (20q13.2-q13.3), which were previously reported to be amplified and overexpressed in ovarian cancers. Some of these genes may be involved in immortalization of HOSE cells and represent crucial premalignant changes in ovarian surface epithelium. Investigation of the involvement of these genes was examined in four pairs of pre-crisis (preimmortalized) and post-crisis (immortalized) HOSE cells. Overexpression of AURKA (Aurora kinase A), also known as BTAK and STK15, by both real time-quantitative polymerase chain reaction (RT-QPCR) and Western blotting was detected in all the four immortalized HOSE cells examined while overexpression of AIB1 and ZNF217 was observed in two of four immortalized HOSE cells examined. Overexpression of TGIF2 and PTPN1 was not significant in our immortalized HOSE cell systems. The degree of overexpression of AURKA was shown to be closely associated with the amplification of chromosome 20q in immortalized HOSE cells. Fluorescence in situ hybridization (FISH) with labeled P1 artificial clone (PAC) confirmed the amplification of the chromosomal region (20q13.2-13.3) where AURKA resides. DNA amplification of AURKA was also confirmed using semi-quantitative PCR. Our study showed that amplification and overexpression of AURKA is a common and significant event during immortalization of HOSE cells and may represent an important premalignant change in ovarian carcinogenesis. Copyright (c) 2005 Wiley-Liss, Inc.

  3. In-gel multiple displacement amplification of long DNA fragments diluted to the single molecule level.

    Science.gov (United States)

    Michikawa, Yuichi; Sugahara, Keisuke; Suga, Tomo; Ohtsuka, Yoshimi; Ishikawa, Kenichi; Ishikawa, Atsuko; Shiomi, Naoko; Shiomi, Tadahiro; Iwakawa, Mayumi; Imai, Takashi

    2008-12-15

    The isolation and multiple genotyping of long individual DNA fragments are needed to obtain haplotype information for diploid organisms. Limiting dilution of sample DNA followed by multiple displacement amplification is a useful technique but is restricted to short (reaction (PCR)-ready form. The haplotypes of seven SNPs spanning 240 kb of the DNA surrounding the human ATM gene region on chromosome 11 were determined for 10 individuals, demonstrating the feasibility of this new method.

  4. Rapid screening method for male DNA by using the loop-mediated isothermal amplification assay.

    Science.gov (United States)

    Kitamura, Masashi; Kubo, Seiji; Tanaka, Jin; Adachi, Tatsushi

    2017-08-12

    Screening for male-derived biological material from collected samples plays an important role in criminal investigations, especially those involving sexual assaults. We have developed a loop-mediated isothermal amplification (LAMP) assay targeting multi-repeat sequences of the Y chromosome for detecting male DNA. Successful amplification occurred with 0.5 ng of male DNA under isothermal conditions of 61 to 67 °C, but no amplification occurred with up to 10 ng of female DNA. Under the optimized conditions, the LAMP reaction initiated amplification within 10 min and amplified for 20 min. The LAMP reaction was sensitive at levels as low as 1-pg male DNA, and a quantitative LAMP assay could be developed because of the strong correlation between the reaction time and the amount of template DNA in the range of 10 pg to 10 ng. Furthermore, to apply the LAMP assay to on-site screening for male-derived samples, we evaluated a protocol using a simple DNA extraction method and a colorimetric intercalating dye that allows detection of the LAMP reaction by evaluating the change in color of the solution. Using this protocol, samples of male-derived blood and saliva stains were processed in approximately 30 min from DNA extraction to detection. Because our protocol does not require much hands-on time or special equipment, this LAMP assay promises to become a rapid and simple screening method for male-derived samples in forensic investigations.

  5. Comparison of the Soil Dynamic Amplification Factor and Soil Amplification by Using Microtremor and MASW Methods Respectively

    Science.gov (United States)

    Tuncel, Aykut; Cevdet Özdag, Özkan; Pamuk, Eren; Akgün, Mustafa

    2017-12-01

    Single Station Microtremor method, which is widely used nowadays, is an effective and easy applicable method. In this study, dynamic amplification factor distributions of the study area were obtained using scenario earthquake parameters with single station microtremor data gathered at 112 points. In addition, a surface wave active method, which is known as MASW (Multichannel Analysis of Surface Waves), was applied at 43 profiles to calculate the soil amplification values. Dynamic amplification factor (DAF), soil amplification, the predominant soil period (PSP), geology and topography data of the study area were analysed together. Dynamic amplification factor and soil amplification values were obtained 2 or higher at about sea level parts of the study area which are generally composed of alluvial units. Additionally, in high altitude regions that are composed of volcanic rocks, relatively lower dynamic amplification factor and soil amplification values were obtained. The minimum amplification value in the study area was 1.15, while the maximum amplification value was 3.05 according to the dynamic amplification results and the soil amplification values were between 1.16 and 3.85 in harmony. It is seen that the obtained DAF values and the soil amplification values calculated from the seismic velocities are very similar to each other numerically and regionally. Because of this, it is concluded that the values of the soil amplification obtained by the MASW method and the calculated DAF values in this study are in harmony with each other. Although the depths of research in these two calculation methods are different from each other, the similarity of the results allows us to arrive at the result of how effective the ground layer is on the amplification. It has a great importance to calculate the amplification values and other dynamic parameters by in situ measurements for a planned plot because geological units can vary even at very short distances in heterogeneously

  6. Communities of arbuscular mycorrhizal fungi in the roots of Pyrus pyrifolia var. culta (Japanese pear) in orchards with variable amounts of soil-available phosphorus.

    Science.gov (United States)

    Yoshimura, Yuko; Ido, Akifumi; Iwase, Koji; Matsumoto, Teruyuki; Yamato, Masahide

    2013-01-01

    We examined the colonization rate and communities of arbuscular mycorrhizal fungi (AMF) in the roots of Pyrus pyrifolia var. culta (Japanese pear) in orchards to investigate the effect of phosphorus (P) fertilization on AMF. Soil cores containing the roots of Japanese pear were collected from 13 orchards in Tottori Prefecture, Japan. Soil-available P in the examined orchards was 75.7 to 1,200 mg kg(-1), showing the extreme accumulation of soil P in many orchards. The AMF colonization rate was negatively correlated with soil-available P (P soil-available P (P fungi may be adapted to high soil-available P conditions. Redundancy analysis showed the significant effects of soil pH, available P in soil, and P content in leaves of P. pyrifolia var. culta trees on AMF distribution. These results suggested that the accumulation of soil-available P affected AMF communities in the roots of Japanese pear in the orchard environment.

  7. Effect of aqueous extracts of alligator pear seed (Persea americana mill) on blood glucose and histopathology of pancreas in alloxan-induced diabetic rats.

    Science.gov (United States)

    Edem, Do; Ekanem, Is; Ebong, Pe

    2009-07-01

    Effects of aqueous extract of alligator pear seed on normal and alloxan-induced diabetic rats were investigated in 6 groups of rats (5 rats per group). Test groups were made diabetic with intra-peritoneal injection of alloxan and treated with 300 mg and 600 mg/kg body weight of alligator pear seed extract. Two non-diabetic groups were also administered with 300 mg and 600 mg/kg body weight extract. The levels of blood glucose were examined in all 6 experimental groups. In diabetic rats, blood glucose levels were significantly reduced (pblood glucose levels were significantly reduced (palligator pear seed may contribute significantly to the reduction of blood glucose levels and can be useful in the treatment of diabetes.

  8. Detection and typing of human papillomavirus in archival cervical cancer specimens by DNA amplification with consensus primers

    NARCIS (Netherlands)

    RESNICK, R. M.; Cornelissen, M. T.; WRIGHT, D. K.; EICHINGER, G. H.; FOX, H. S.; ter Schegget, J.; MANOS, M. M.

    1990-01-01

    We developed a polymerase chain reaction DNA amplification system using two distinct consensus oligonucleotide primer sets for the improved detection and typing of a broad spectrum of human genital papillomavirus (HPV) sequences, including those of novel viruses. The system incorporates one primer

  9. Study on Pollination and Selection of the Most Suitable Pollinizers for Commercial Pear Cultivars (Pyrus communis L. in Iran

    Directory of Open Access Journals (Sweden)

    Tatari Maryam

    2017-12-01

    Full Text Available The cultivated pear is an economically important fruit tree species of genus Pyrus in which often gametophytic self-incompatibility occurs. Therefore, this species need to be pollinated by cross-compatible cultivars that bloom in the same time. Selection of appropriate pollinizers for pear cultivars is very important to produce commercial yield. ‘Sebri’, ‘ Shahmiveh’ and ‘Natanzi’ are the best commercial cultivars in Iran, but the lack of a suitable source of pollen can reduce productivity. In order to select the most suitable pollinizer for these pear cvs, an experiment was conducted in which they were considered as pollen recipients and ‘Coscia’, ‘Bartlett’ and ‘Sardroud’ along with ‘Sebri’, ‘Shahmiveh’ and ‘Natanzi’ were evaluated as pollen donors. This research was conducted as a factorial experiment in randomized complete block design for four years. Recipient and donor cultivars had almost overlapping flowering time. The flower buds on selected branches were emasculated at balloon stage and then were counted and isolated with cotton tissue bags. Pollen grains of these pollinizers were collected in the laboratory. Isolation bags were taken off from the branches and emasculated flowers were pollinated with pollen grains of listed pollinizers during receptibility of stigma. The number of pollinated flowers was counted, and branches were covered again with the bags. The results showed that for ‘Sebri’ the best pollinizer was ‘Coscia’ with 5.7% fruit set, for ‘Shahmiveh’ ‘Bartlett’ cv. with 5.8% of fruit set and for ‘Natanzi’, ‘Shahmiveh’ with 5.5% of fruit set.

  10. Dormancy-Associated MADS-Box (DAM) and the Abscisic Acid Pathway Regulate Pear Endodormancy Through a Feedback Mechanism.

    Science.gov (United States)

    Tuan, Pham Anh; Bai, Songling; Saito, Takanori; Ito, Akiko; Moriguchi, Takaya

    2017-08-01

    In the pear 'Kosui' (Pyrus pyrifolia Nakai), the dormancy-associated MADS-box (PpDAM1 = PpMADS13-1) gene has been reported to play an essential role in bud endodormancy. Here, we found that PpDAM1 up-regulated expression of 9-cis-epoxycarotenoid dioxygenase (PpNCED3), which is a rate-limiting gene for ABA biosynthesis. Transient assays with a dual luciferase reporter system (LUC assay) and electrophoretic mobility shift assay (EMSA) showed that PpDAM1 activated PpNCED3 expression by binding to the CArG motif in the PpNCED3 promoter. PpNCED3 expression was increased toward endodormancy release in lateral flower buds of 'Kosui', which is consistent with the induced levels of ABA, its catabolism (ABA 8'-hydroxylase) and signaling genes (type 2C protein phosphatase genes and SNF1-related protein kinase 2 genes). In addition, we found that an ABA response element (ABRE)-binding transcription factor, PpAREB1, exhibiting high expression concomitant with endodormancy release, bound to three ABRE motifs in the promoter region of PpDAM1 and negatively regulated its activity. Taken together, our results suggested a feedback regulation between PpDAM1 and the ABA metabolism and signaling pathway during endodormancy of pear. This first evidence of an interaction between a DAM and ABA biosynthesis in vitro will provide further insights into bud endodormancy regulatory mechanisms of deciduous trees including pear. © The Author 2017. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  11. Rehabilitation of Degraded Rangeland in Drylands by Prickly Pear (Opuntia ficus-indica L. Plantations: Effect on Soil and Spontaneous Vegetation

    Directory of Open Access Journals (Sweden)

    Souad Neffar

    2013-12-01

    Full Text Available In arid and semi-arid lands, the spiny prickly pear (Opuntia ficus-indica is an outstanding plant for soil conservation and restoration. To determine the role of Opuntia ficus-indica on vegetation recovery process in desertified areas of Southern Tebessa (Northeast Algeria, we investigated the effect of prickly pear plantation age and some soil properties (grain size, pH, electrical conductivity, organic matter, total nitrogen, available phosphorus, and CaCO3 equivalents on native plant community. Vegetation cover and plant diversity were assessed by calculating the number of individual plants (N, species richness (S, their ratio (N/S, Shannon index, and Evenness in prickly pear plantation plots of different ages (control, 5 and 20 years. Even if surveyed soil parameters did not differ significantly among O. ficus-indica plantations, results of ANOVA testing the effect of Opuntia plantations on native vegetation traits revealed significant variation for plant abundance (P < 0.0001, N/S ratio (P = 0.003 and vegetation cover (P < 0.0001. Vegetation cover differed significantly with both prickly-pear plantation age (P = 0.031 and seasons (P = 0.019. Tukey's tests revealed that all vegetation traits were significantly higher on prickly pear plantations than in control plots. Multiple comparisons also showed that plant abundance, N/S ratio and vegetation cover were significantly different between both young and old plantations and the controls. Prickly pear cultures facilitated the colonization and development of herbaceous species by ameliorating the severe environmental conditions. In conclusion, the facilitative effect of O. ficus-indica has been clearly demonstrated for both abundance and cover of native vegetation.

  12. Detection of the 35S promoter in transgenic maize via various isothermal amplification techniques: a practical approach.

    Science.gov (United States)

    Zahradnik, Celine; Kolm, Claudia; Martzy, Roland; Mach, Robert L; Krska, Rudolf; Farnleitner, Andreas H; Brunner, Kurt

    2014-11-01

    In 2003 the European Commission introduced a 0.9% threshold for food and feed products containing genetically modified organism (GMO)-derived components. For commodities containing GMO contents higher than this threshold, labelling is mandatory. To provide a DNA-based rapid and simple detection method suitable for high-throughput screening of GMOs, several isothermal amplification approaches for the 35S promoter were tested: strand displacement amplification, nicking-enzyme amplification reaction, rolling circle amplification, loop-mediated isothermal amplification (LAMP) and helicase-dependent amplification (HDA). The assays developed were tested for specificity in order to distinguish between samples containing genetically modified (GM) maize and non-GM maize. For those assays capable of this discrimination, tests were performed to determine the lower limit of detection. A false-negative rate was determined to rule out whether GMO-positive samples were incorrectly classified as GMO-negative. A robustness test was performed to show reliable detection independent from the instrument used for amplification. The analysis of three GM maize lines showed that only LAMP and HDA were able to differentiate between the GMOs MON810, NK603, and Bt11 and non-GM maize. Furthermore, with the HDA assay it was possible to realize a detection limit as low as 0.5%. A false-negative rate of only 5% for 1% GM maize for all three maize lines shows that HDA has the potential to be used as an alternative strategy for the detection of transgenic maize. All results obtained with the LAMP and HDA assays were compared with the results obtained with a previously reported real-time PCR assay for the 35S promoter in transgenic maize. This study presents two new screening assays for detection of the 35S promoter in transgenic maize by applying the isothermal amplification approaches HDA and LAMP.

  13. An integrated portable hand-held analyser for real-time isothermal nucleic acid amplification

    Energy Technology Data Exchange (ETDEWEB)

    Smith, Matthew C. [College of Marine Science, University of South Florida, St Petersburg, FL (United States)], E-mail: msmith@marine.usf.edu; Steimle, George; Ivanov, Stan; Holly, Mark; Fries, David P. [College of Marine Science, University of South Florida, St Petersburg, FL (United States)

    2007-08-29

    A compact hand-held heated fluorometric instrument for performing real-time isothermal nucleic acid amplification and detection is described. The optoelectronic instrument combines a Printed Circuit Board/Micro Electro Mechanical Systems (PCB/MEMS) reaction detection/chamber containing an integrated resistive heater with attached miniature LED light source and photo-detector and a disposable glass waveguide capillary to enable a mini-fluorometer. The fluorometer is fabricated and assembled in planar geometry, rolled into a tubular format and packaged with custom control electronics to form the hand-held reactor. Positive or negative results for each reaction are displayed to the user using an LED interface. Reaction data is stored in FLASH memory for retrieval via an in-built USB connection. Operating on one disposable 3 V lithium battery >12, 60 min reactions can be performed. Maximum dimensions of the system are 150 mm (h) x 48 mm (d) x 40 mm (w), the total instrument weight (with battery) is 140 g. The system produces comparable results to laboratory instrumentation when performing a real-time nucleic acid sequence-based amplification (NASBA) reaction, and also displayed comparable precision, accuracy and resolution to laboratory-based real-time nucleic acid amplification instrumentation. A good linear response (R{sup 2} = 0.948) to fluorescein gradients ranging from 0.5 to 10 {mu}M was also obtained from the instrument indicating that it may be utilized for other fluorometric assays. This instrument enables an inexpensive, compact approach to in-field genetic screening, providing results comparable to laboratory equipment with rapid user feedback as to the status of the reaction.

  14. An integrated portable hand-held analyser for real-time isothermal nucleic acid amplification

    International Nuclear Information System (INIS)

    Smith, Matthew C.; Steimle, George; Ivanov, Stan; Holly, Mark; Fries, David P.

    2007-01-01

    A compact hand-held heated fluorometric instrument for performing real-time isothermal nucleic acid amplification and detection is described. The optoelectronic instrument combines a Printed Circuit Board/Micro Electro Mechanical Systems (PCB/MEMS) reaction detection/chamber containing an integrated resistive heater with attached miniature LED light source and photo-detector and a disposable glass waveguide capillary to enable a mini-fluorometer. The fluorometer is fabricated and assembled in planar geometry, rolled into a tubular format and packaged with custom control electronics to form the hand-held reactor. Positive or negative results for each reaction are displayed to the user using an LED interface. Reaction data is stored in FLASH memory for retrieval via an in-built USB connection. Operating on one disposable 3 V lithium battery >12, 60 min reactions can be performed. Maximum dimensions of the system are 150 mm (h) x 48 mm (d) x 40 mm (w), the total instrument weight (with battery) is 140 g. The system produces comparable results to laboratory instrumentation when performing a real-time nucleic acid sequence-based amplification (NASBA) reaction, and also displayed comparable precision, accuracy and resolution to laboratory-based real-time nucleic acid amplification instrumentation. A good linear response (R 2 = 0.948) to fluorescein gradients ranging from 0.5 to 10 μM was also obtained from the instrument indicating that it may be utilized for other fluorometric assays. This instrument enables an inexpensive, compact approach to in-field genetic screening, providing results comparable to laboratory equipment with rapid user feedback as to the status of the reaction

  15. Light amplification by seeded Kerr instability

    Science.gov (United States)

    Vampa, G.; Hammond, T. J.; Nesrallah, M.; Naumov, A. Yu.; Corkum, P. B.; Brabec, T.

    2018-02-01

    Amplification of femtosecond laser pulses typically requires a lasing medium or a nonlinear crystal. In either case, the chemical properties of the lasing medium or the momentum conservation in the nonlinear crystal constrain the frequency and the bandwidth of the amplified pulses. We demonstrate high gain amplification (greater than 1000) of widely tunable (0.5 to 2.2 micrometers) and short (less than 60 femtosecond) laser pulses, up to intensities of 1 terawatt per square centimeter, by seeding the modulation instability in an Y3Al5O12 crystal pumped by femtosecond near-infrared pulses. Our method avoids constraints related to doping and phase matching and therefore can occur in a wider pool of glasses and crystals even at far-infrared frequencies and for single-cycle pulses. Such amplified pulses are ideal to study strong-field processes in solids and highly excited states in gases.

  16. Parametric nanomechanical amplification at very high frequency.

    Science.gov (United States)

    Karabalin, R B; Feng, X L; Roukes, M L

    2009-09-01

    Parametric resonance and amplification are important in both fundamental physics and technological applications. Here we report very high frequency (VHF) parametric resonators and mechanical-domain amplifiers based on nanoelectromechanical systems (NEMS). Compound mechanical nanostructures patterned by multilayer, top-down nanofabrication are read out by a novel scheme that parametrically modulates longitudinal stress in doubly clamped beam NEMS resonators. Parametric pumping and signal amplification are demonstrated for VHF resonators up to approximately 130 MHz and provide useful enhancement of both resonance signal amplitude and quality factor. We find that Joule heating and reduced thermal conductance in these nanostructures ultimately impose an upper limit to device performance. We develop a theoretical model to account for both the parametric response and nonequilibrium thermal transport in these composite nanostructures. The results closely conform to our experimental observations, elucidate the frequency and threshold-voltage scaling in parametric VHF NEMS resonators and sensors, and establish the ultimate sensitivity limits of this approach.

  17. In Vitro Propagation of Three Moroccan Prickly Pear Cactus Opuntia and Plant Establishment in Soil

    Directory of Open Access Journals (Sweden)

    Aissam EL FINTI

    2013-02-01

    Full Text Available Opuntia is one of the most widespread cacti, primarily due to their edible fruit and vegetable mass used as feed. The high demand for young plants of Opuntia made it necessary to find a rapid method of multiplication of the cactus, the safest method consisting in vitro micropropagation of species belonging to this genus. With aim of large production of plant material, a propagation system of three important prickly pear cactus cultivar (Opuntia ficus-indica in Morocco was developed. Segments of healthy young cladode (containing one areole were cultivated in Murashige and Skoog medium (MS containing adenine sulfate (40 mg/1, monosodium phosphate (50 mg/l, sucrose (50 g/l, phytagel (0.3% and benzyladenine (BA at 22.2 μM, to start the process of micropropagation. In vitro-developed shoots from areoles were used as secondary explants to induce shoot development in the MS medium with 5 mg/l of BA. All of the three studied cultivars showed an important multiplication rate in this medium. ‘Sidi Ifni M’ (‘Moussa’ cultivar shows the greatest number of shoots followed by ‘Sidi Ifni A’ (‘Aissa’ and ‘Delahia’ 17.26, 14.12 and 12.13 respectively. Rooting of in vitro-generated shoots was achieved most efficiently on half-strength MS basal medium supplemented with 0.5 mg/l of indole-3-butyric acid (IBA or IAA. Rooting frequencies were in the range from 95 to 100% and the highest mean number of root (19.1 was obtained with IBA for ‘Delahia’ cultivar. All micropropagated plants were transferred to greenhouse and all of them survived acclimatization process and showed good overall growth.

  18. Conservation of apple and pear juice concentrates. Synergic effect of heat and radiation

    International Nuclear Information System (INIS)

    Kaupert, N.L.; Lescano, H.G.; Kotliar, N.

    1981-01-01

    This paper aims at assessing the feasibility for conserving apple and pear-juice concentrates through the synergic action of heat and radiation. The material was packed in sterile 100-μm polyethylene bags and, after the treatment was applied, the resulting fractions were stored under room temperature (25 0 C+-1 0 C). The temperature applied to the samples before irradiation was 50 0 C during 10 minutes and the doses were 100, 200, 300 and 400 krad. For such purposes, a 60 Co 165-krad/h source was used, located in a mobile irradiator. Periodical microbiological, chemical and organoleptic controls of the food were performed on both control and on irradiated samples, with or without heat. A single alterating microorganism was isolated from all the samples, which was featured as Saccharomyces rouxii. Adopting the temperature as an application variable and the absorbed dose as a constant, the above osmophilic yeast is considerably more sensitive to radiations when it is suspended in a 50% sucrose solution, after the latter was submitted to a 50 0 C temperature treatment. It has been proved that 72.5 krad are needed to attain the reduction of a logarithmic cycle in the Saccharomyces rouxii population irradiated at room temperature, while 36 krad are needed if the sample has been previously heated to 50 0 C for 10 minutes. An attempt was made to apply the synergism of such process to the juice concentrate. Below 50 0 C associated with 400 krad gamma radiation, a total inactivation took place in the Saccharomyces rouxii during the 150 days under analysis. Colour changes were detected in the concentrate; however, the acceptability features for consumption remained at a normal value (level 5) in a hedonic scale of 7 points. (author)

  19. Hormonal Involvement in Breast Cancer Gene Amplification

    Science.gov (United States)

    2010-10-01

    been shown to induce DN A amplification in yeast (Gopalakrishnan et al., 2001; Nguy en et al., 2001; Green et al., 2006) an d increased Cdt1 results in...re-replication in human cells (Dorn et al., 2008). The N- terminus of Cdt1 is important for re-replication, perhaps through interactions with PCNA...evolution of a cancer genome. Genome Res. (Epub. Dec. 3, 2008). Harris TD, Buzby PR, Babcock H, Beer E, Bowers J, Bras lavsky I, Causey M

  20. Fast amplification system for gamma spectroscopy

    International Nuclear Information System (INIS)

    Jesus, E.F.O.; Lopes, R.T.

    1992-01-01

    An amplification system for gamma spectroscopy with high counting rates was developed. The system was constructed with operational amplifiers, and tested and compared with ORTEC conventional system, using Iridium-192 as source of 9,25 x 10 1 0 Bq of activity and NaI (Tl) detector. The constructed system showed a better performance in relation to efficiency and resolution parameters, tested before. (C.G.C.)

  1. Optimized thermal amplification in a radiative transistor

    Energy Technology Data Exchange (ETDEWEB)

    Prod' homme, Hugo; Ordonez-Miranda, Jose; Ezzahri, Younes, E-mail: younes.ezzahri@univ-poitiers.fr; Drevillon, Jeremie; Joulain, Karl [Institut Pprime, CNRS, Université de Poitiers, ISAE-ENSMA, F-86962 Futuroscope Chasseneuil (France)

    2016-05-21

    The thermal performance of a far-field radiative transistor made up of a VO{sub 2} base in between a blackbody collector and a blackbody emitter is theoretically studied and optimized. This is done by using the grey approximation on the emissivity of VO{sub 2} and deriving analytical expressions for the involved heat fluxes and transistor amplification factor. It is shown that this amplification factor can be maximized by tuning the base temperature close to its critical one, which is determined by the temperature derivative of the VO{sub 2} emissivity and the equilibrium temperatures of the collector and emitter. This maximization is the result of the presence of two bi-stable temperatures appearing during the heating and cooling processes of the VO{sub 2} base and enables a thermal switching (temperature jump) characterized by a sizeable variation of the collector-to-base and base-to-emitter heat fluxes associated with a slight change of the applied power to the base. This switching effect leads to the optimization of the amplification factor and therefore it could be used for thermal modulation purposes.

  2. Systematic evaluation of bias in microbial community profiles induced by whole genome amplification.

    Science.gov (United States)

    Direito, Susana O L; Zaura, Egija; Little, Miranda; Ehrenfreund, Pascale; Röling, Wilfred F M

    2014-03-01

    Whole genome amplification methods facilitate the detection and characterization of microbial communities in low biomass environments. We examined the extent to which the actual community structure is reliably revealed and factors contributing to bias. One widely used [multiple displacement amplification (MDA)] and one new primer-free method [primase-based whole genome amplification (pWGA)] were compared using a polymerase chain reaction (PCR)-based method as control. Pyrosequencing of an environmental sample and principal component analysis revealed that MDA impacted community profiles more strongly than pWGA and indicated that this related to species GC content, although an influence of DNA integrity could not be excluded. Subsequently, biases by species GC content, DNA integrity and fragment size were separately analysed using defined mixtures of DNA from various species. We found significantly less amplification of species with the highest GC content for MDA-based templates and, to a lesser extent, for pWGA. DNA fragmentation also interfered severely: species with more fragmented DNA were less amplified with MDA and pWGA. pWGA was unable to amplify low molecular weight DNA (microbial communities in low-biomass environments and for currently planned astrobiological missions to Mars. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

  3. New Fpg probe chemistry for direct detection of recombinase polymerase amplification on lateral flow strips.

    Science.gov (United States)

    Powell, Michael L; Bowler, Frank R; Martinez, Aurore J; Greenwood, Catherine J; Armes, Niall; Piepenburg, Olaf

    2018-02-15

    Rapid, cost-effective and sensitive detection of nucleic acids has the ability to improve upon current practices employed for pathogen detection in diagnosis of infectious disease and food testing. Furthermore, if assay complexity can be reduced, nucleic acid amplification tests could be deployed in resource-limited and home use scenarios. In this study, we developed a novel Fpg (Formamidopyrimidine DNA glycosylase) probe chemistry, which allows lateral flow detection of amplification in undiluted recombinase polymerase amplification (RPA) reactions. The prototype nucleic acid lateral flow chemistry was applied to a human genomic target (rs1207445), Campylobacter jejuni 16S rDNA and two genetic markers of the important food pathogen E. coli O157:H7. All four assays have an analytical sensitivity between 10 and 100 copies DNA per amplification. Furthermore, the assay is performed with fewer hands-on steps than using the current RPA Nfo lateral flow method as dilution of amplicon is not required for lateral flow analysis. Due to the simplicity of the workflow, we believe that the lateral flow chemistry for direct detection could be readily adapted to a cost-effective single-use consumable, ideal for use in non-laboratory settings. Copyright © 2017. Published by Elsevier Inc.

  4. Generation of sequence signatures from DNA amplification fingerprints with mini-hairpin and microsatellite primers.

    Science.gov (United States)

    Caetano-Anollés, G; Gresshoff, P M

    1996-06-01

    DNA amplification fingerprinting (DAF) with mini-hairpins harboring arbitrary "core" sequences at their 3' termini were used to fingerprint a variety of templates, including PCR products and whole genomes, to establish genetic relationships between plant tax at the interspecific and intraspecific level, and to identify closely related fungal isolates and plant accessions. No correlation was observed between the sequence of the arbitrary core, the stability of the mini-hairpin structure and DAF efficiency. Mini-hairpin primers with short arbitrary cores and primers complementary to simple sequence repeats present in microsatellites were also used to generate arbitrary signatures from amplification profiles (ASAP). The ASAP strategy is a dual-step amplification procedure that uses at least one primer in each fingerprinting stage. ASAP was able to reproducibly amplify DAF products (representing about 10-15 kb of sequence) following careful optimization of amplification parameters such as primer and template concentration. Avoidance of primer sequences partially complementary to DAF product termini was necessary in order to produce distinct fingerprints. This allowed the combinatorial use of oligomers in nucleic acid screening, with numerous ASAP fingerprinting reactions based on a limited number of primer sequences. Mini-hairpin primers and ASAP analysis significantly increased detection of polymorphic DNA, separating closely related bermudagrass (Cynodon) cultivars and detecting putatively linked markers in bulked segregant analysis of the soybean (Glycine max) supernodulation (nitrate-tolerant symbiosis) locus.

  5. Development of an Automated Microfluidic System for DNA Collection, Amplification, and Detection of Pathogens

    Energy Technology Data Exchange (ETDEWEB)

    Hagan, Bethany S.; Bruckner-Lea, Cynthia J.

    2002-12-01

    This project was focused on developing and testing automated routines for a microfluidic Pathogen Detection System. The basic pathogen detection routine has three primary components; cell concentration, DNA amplification, and detection. In cell concentration, magnetic beads are held in a flow cell by an electromagnet. Sample liquid is passed through the flow cell and bacterial cells attach to the beads. These beads are then released into a small volume of fluid and delivered to the peltier device for cell lysis and DNA amplification. The cells are lysed during initial heating in the peltier device, and the released DNA is amplified using polymerase chain reaction (PCR) or strand displacement amplification (SDA). Once amplified, the DNA is then delivered to a laser induced fluorescence detection unit in which the sample is detected. These three components create a flexible platform that can be used for pathogen detection in liquid and sediment samples. Future developments of the system will include on-line DNA detection during DNA amplification and improved capture and release methods for the magnetic beads during cell concentration.

  6. The role of DNA amplification and cultural growth in complicated acute appendicitis

    Directory of Open Access Journals (Sweden)

    Francesca Tocchioni

    2016-09-01

    Full Text Available Bacterial growth of peritoneal fluid specimens obtained during surgical procedures for acute appendicitis may be useful to optimize further antibiotic therapy in complicated cases. DNA amplification represents a fast technique to detect microbial sequences. We aimed to compare the potential of DNA amplification versus traditional bacterial growth culture highlighting advantages and drawbacks in a surgical setting. Peritoneal fluid specimens were collected during surgery from 36 children who underwent appendectomy between May and December 2012. Real-time polymerase chain reaction (RT-PCR and cultures were performed on each sample. RT-PCR showed an amplification of 16S in 18/36 samples, Escherichia coli (in 7 cases, Pseudomonas aeruginosa (3, Fusobacterium necrophorum (3, Adenovirus (2, E.coli (1, Klebsiella pneumoniae (1, Serratia marcescens/Enterobacter cloacae (1. Bacterial growth was instead observed only in four patients (3 E.coli and 1 P.aeruginosa and Bacteroides ovatus. Preoperative C-reactive protein and inflammation degree, the most reliable indicators of bacterial translocation, were elevated as expected. DNA amplification was a quick and useful method to detect pathogens and it was even more valuable in detecting aggressive pathogens such as anaerobes, difficult to preserve in biological cultures; its drawbacks were the lack of biological growths and of antibiograms. In our pilot study RT-PCR and cultures did not influence the way patients were treated.

  7. HER-2 amplification in tubular carcinoma of the breast.

    Science.gov (United States)

    Oakley, Gerard J; Tubbs, Raymond R; Crowe, Joseph; Sebek, Bruce; Budd, G Thomas; Patrick, Rebecca J; Procop, Gary W

    2006-07-01

    The prognostic and therapeutic implications of HER-2 gene amplification and estrogen and progesterone receptor status in breast cancer are well described. To address the relative paucity of information concerning HER-2 amplification for tubular carcinomas, we assessed the frequency of gene amplification in 55 tubular carcinomas of the breast from 54 patients, 5 of which had axillary node metastases. The HER-2 gene copy number was assessed by fluorescence in situ hybridization for the majority of tumors analyzed, whereas estrogen and progesterone receptor status was achieved by immunohistochemical analysis. HER-2 gene amplification was not observed in any of the tumors examined, and most were estrogen receptor-positive. This HER-2 gene amplification frequency was significantly lower than the frequency of gene amplification previously reported for all invasive ductal carcinoma of no special type (P < .01). HER-2 gene amplification likely occurs infrequently, or not at all, in tubular carcinomas of the breast, whereas most express estrogen receptors.

  8. Social Amplification of Risk and Crisis Communication Planing - Case Study

    Science.gov (United States)

    Stanciugelu, I.; Frunzaru, V.; Armas, I.; Duntzer, A.; Stan, S.

    2012-04-01

    Risk management has become a dominant concern of public policy and the ability of government to anticipate the strength and focus of public concerns remains weak. The Social Amplification of Risk Framework (SARF) was designed to assist in this endeavor. It aims to facilitate a greater understanding of the social processes that can mediate between a hazard event and its consequences. SARF identifies categories of mediator/moderator that intervene between risk event and its consequences and suggests a causal and temporal sequence in which they act. Information flows first through various sources and then channels, triggering social stations of amplification, initiating individual station of amplification and precipitating behavioral reactions. The International Risk Governance Council Framework is an interdisciplinary and multilevel approach, linking risk management and risk assessment sphere through communication. This study aims to identify categories of mediator/moderator that intervene between the risk event and its consequences, using a survey on earthquake risk perception addressing population of Bucharest city. Romania has a unique seismic profile in Europe, being the country with the biggest surface affected in case of a serious earthquake. Considering the development of the urban area that took place in the last two decades and the growing number of inhabitants, Bucharest is the largest city in Romania and is exposed to extensive damages in case of an earthquake. The sociological survey has been conducted in December 2009 on a representative sample of the Bucharest population aged 18 and over (N=1376) using one stage sampling design. We used a stratified sample method shearing the investigated populations in six layers according to the six sectors of Bucharest. The respondents were selected using random digit dialling method (RDD) and the questionnaires were administered by research staff with computer assisted telephone interviewing method (CATI). The

  9. PpHB22, a member of HD-Zip proteins, activates PpDAM1 to regulate bud dormancy transition in 'Suli' pear (Pyrus pyrifolia White Pear Group).

    Science.gov (United States)

    Yang, Qinsong; Niu, Qingfeng; Li, Jianzhao; Zheng, Xiaoyan; Ma, Yunjing; Bai, Songling; Teng, Yuanwen

    2018-06-01

    Homeodomain-leucine zipper (HD-Zip) proteins, which form one of the largest and most diverse families, regulate many biological processes in plants, including differentiation, flowering, vascular development, and stress signaling. Abscisic acid (ABA) has been proved to be one of the key regulators of bud dormancy and to influence several HD-Zip genes expression. However, the role of HD-Zip genes in regulating bud dormancy remains unclear. We identified 47 pear (P. pyrifolia White Pear Group) HD-Zip genes, which were classified into four subfamilies (HD-Zip I-IV). We further revealed that gene expression levels of some HD-Zip members were closely related to ABA concentrations in flower buds during dormancy transition. Exogenous ABA treatment confirmed that PpHB22 and several other HD-Zip genes responded to ABA. Yeast one-hybrid and dual luciferase assay results combining subcellular localization showed that PpHB22 was present in nucleus and directly induced PpDAM1 (dormancy associated MADS-box 1) expression. Thus, PpHB22 is a negative regulator of plant growth associated with the ABA response pathway and functions upstream of PpDAM1. These findings enrich our understanding of the function of HD-Zip genes related to the bud dormancy transition. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  10. A large ungated TPC with GEM amplification

    Science.gov (United States)

    Berger, M.; Ball, M.; Fabbietti, L.; Ketzer, B.; Arora, R.; Beck, R.; Böhmer, F. V.; Chen, J.-C.; Cusanno, F.; Dørheim, S.; García, F.; Hehner, J.; Herrmann, N.; Höppner, C.; Kaiser, D.; Kis̆, M.; Kleipa, V.; Konorov, I.; Kunkel, J.; Kurz, N.; Leifels, Y.; Müllner, P.; Münzer, R.; Neubert, S.; Rauch, J.; Schmidt, C. J.; Schmitz, R.; Soyk, D.; Vandenbroucke, M.; Voss, B.; Walther, D.; Zmeskal, J.

    2017-10-01

    A Time Projection Chamber (TPC) is an ideal device for the detection of charged particle tracks in a large volume covering a solid angle of almost 4 π. The high density of hits on a given particle track facilitates the task of pattern recognition in a high-occupancy environment and in addition provides particle identification by measuring the specific energy loss for each track. For these reasons, TPCs with Multiwire Proportional Chamber (MWPC) amplification have been and are widely used in experiments recording heavy-ion collisions. A significant drawback, however, is the large dead time of the order of 1 ms per event generated by the use of a gating grid, which is mandatory to prevent ions created in the amplification region from drifting back into the drift volume, where they would severely distort the drift path of subsequent tracks. For experiments with higher event rates this concept of a conventional TPC operating with a triggered gating grid can therefore not be applied without a significant loss of data. A continuous readout of the signals is the more appropriate way of operation. This, however, constitutes a change of paradigm with considerable challenges to be met concerning the amplification region, the design and bandwidth of the readout electronics, and the data handling. A mandatory prerequisite for such an operation is a sufficiently good suppression of the ion backflow from the avalanche region, which otherwise limits the tracking and particle identification capabilities of such a detector. Gas Electron Multipliers (GEM) are a promising candidate to combine excellent spatial resolution with an intrinsic suppression of ions. In this paper we describe the design, construction and the commissioning of a large TPC with GEM amplification and without gating grid (GEM-TPC). The design requirements have driven innovations in the construction of a light-weight field-cage, a supporting media flange, the GEM amplification and the readout system, which are

  11. Towards rapid prototyped convective microfluidic DNA amplification platform

    Science.gov (United States)

    Ajit, Smrithi; Praveen, Hemanth Mithun; Puneeth, S. B.; Dave, Abhishek; Sesham, Bharat; Mohan, K. N.; Goel, Sanket

    2017-02-01

    Today, Polymerase Chain Reaction (PCR) based DNA amplification plays an indispensable role in the field of biomedical research. Its inherent ability to exponentially amplify sample DNA has proven useful for the identification of virulent pathogens like those causing Multiple Drug-Resistant Tuberculosis (MDR-TB). The intervention of Microfluidics technology has revolutionized the concept of PCR from being a laborious and time consuming process into one that is faster, easily portable and capable of being multifunctional. The Microfluidics based PCR outweighs its traditional counterpart in terms of flexibility of varying reaction rate, operation simplicity, need of a fraction of volume and capability of being integrated with other functional elements. The scope of the present work involves the development of a real-time continuous flow microfluidic device, fabricated by 3D printing-governed rapid prototyping method, eventually leading to an automated and robust platform to process multiple DNA samples for detection of MDRTB-associated mutations. The thermal gradient characteristic to the PCR process is produced using peltier units appropriate to the microfluidic environment fully monitored and controlled by a low cost controller driven by a Data Acquisition System. The process efficiency achieved in the microfluidic environment in terms of output per cycle is expected to be on par with the traditional PCR and capable of earning the additional advantages of being faster and minimizing the handling.

  12. Integrated Microfluidic Nucleic Acid Isolation, Isothermal Amplification, and Amplicon Quantification

    Directory of Open Access Journals (Sweden)

    Michael G. Mauk

    2015-10-01

    Full Text Available Microfluidic components and systems for rapid (<60 min, low-cost, convenient, field-deployable sequence-specific nucleic acid-based amplification tests (NAATs are described. A microfluidic point-of-care (POC diagnostics test to quantify HIV viral load from blood samples serves as a representative and instructive example to discuss the technical issues and capabilities of “lab on a chip” NAAT devices. A portable, miniaturized POC NAAT with performance comparable to conventional PCR (polymerase-chain reaction-based tests in clinical laboratories can be realized with a disposable, palm-sized, plastic microfluidic chip in which: (1 nucleic acids (NAs are extracted from relatively large (~mL volume sample lysates using an embedded porous silica glass fiber or cellulose binding phase (“membrane” to capture sample NAs in a flow-through, filtration mode; (2 NAs captured on the membrane are isothermally (~65 °C amplified; (3 amplicon production is monitored by real-time fluorescence detection, such as with a smartphone CCD camera serving as a low-cost detector; and (4 paraffin-encapsulated, lyophilized reagents for temperature-activated release are pre-stored in the chip. Limits of Detection (LOD better than 103 virons/sample can be achieved. A modified chip with conduits hosting a diffusion-mode amplification process provides a simple visual indicator to readily quantify sample NA template. In addition, a companion microfluidic device for extracting plasma from whole blood without a centrifuge, generating cell-free plasma for chip-based molecular diagnostics, is described. Extensions to a myriad of related applications including, for example, food testing, cancer screening, and insect genotyping are briefly surveyed.

  13. Loop mediated isothermal amplification: An innovative gene amplification technique for animal diseases

    Directory of Open Access Journals (Sweden)

    Pravas Ranjan Sahoo

    2016-05-01

    Full Text Available India being a developing country mainly depends on livestock sector for its economy. However, nowadays, there is emergence and reemergence of more transboundary animal diseases. The existing diagnostic techniques are not so quick and with less specificity. To reduce the economy loss, there should be a development of rapid, reliable, robust diagnostic technique, which can work with high degree of sensitivity and specificity. Loop mediated isothermal amplification assay is a rapid gene amplification technique that amplifies nucleic acid under an isothermal condition with a set of designed primers spanning eight distinct sequences of the target. This assay can be used as an emerging powerful, innovative gene amplification diagnostic tool against various pathogens of livestock diseases. This review is to highlight the basic concept and methodology of this assay in livestock disease.

  14. Loop mediated isothermal amplification: An innovative gene amplification technique for animal diseases.

    Science.gov (United States)

    Sahoo, Pravas Ranjan; Sethy, Kamadev; Mohapatra, Swagat; Panda, Debasis

    2016-05-01

    India being a developing country mainly depends on livestock sector for its economy. However, nowadays, there is emergence and reemergence of more transboundary animal diseases. The existing diagnostic techniques are not so quick and with less specificity. To reduce the economy loss, there should be a development of rapid, reliable, robust diagnostic technique, which can work with high degree of sensitivity and specificity. Loop mediated isothermal amplification assay is a rapid gene amplification technique that amplifies nucleic acid under an isothermal condition with a set of designed primers spanning eight distinct sequences of the target. This assay can be used as an emerging powerful, innovative gene amplification diagnostic tool against various pathogens of livestock diseases. This review is to highlight the basic concept and methodology of this assay in livestock disease.

  15. Comparison of polymerase chain reaction (PCR) and loop-mediated ...

    African Journals Online (AJOL)

    Comparison of polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) for diagnosis of Fusarium solani in human immunodeficiency virus (HIV) positive patients. ... The test was carried out in 1 h reaction at 65°C in a heater block. The specificity of the test was 100% and its sensitivity was a ...

  16. Silicon-based sleeve devices for chemical reactions

    Science.gov (United States)

    Northrup, M. Allen; Mariella, Jr., Raymond P.; Carrano, Anthony V.; Balch, Joseph W.

    1996-01-01

    A silicon-based sleeve type chemical reaction chamber that combines heaters, such as doped polysilicon for heating, and bulk silicon for convection cooling. The reaction chamber combines a critical ratio of silicon and silicon nitride to the volume of material to be heated (e.g., a liquid) in order to provide uniform heating, yet low power requirements. The reaction chamber will also allow the introduction of a secondary tube (e.g., plastic) into the reaction sleeve that contains the reaction mixture thereby alleviating any potential materials incompatibility issues. The reaction chamber may be utilized in any chemical reaction system for synthesis or processing of organic, inorganic, or biochemical reactions, such as the polymerase chain reaction (PCR) and/or other DNA reactions, such as the ligase chain reaction, which are examples of a synthetic, thermal-cycling-based reaction. The reaction chamber may also be used in synthesis instruments, particularly those for DNA amplification and synthesis.

  17. Genome-wide identification and comparative analysis of the cation proton antiporters family in pear and four other Rosaceae species.

    Science.gov (United States)

    Zhou, Hongsheng; Qi, Kaijie; Liu, Xing; Yin, Hao; Wang, Peng; Chen, Jianqing; Wu, Juyou; Zhang, Shaoling

    2016-08-01

    The monovalent cation proton antiporters (CPAs) play essential roles in plant nutrition, development, and signal transduction by regulating ion and pH homeostasis of the cell. The CPAs of plants include the Na(+)/H(+) exchanger, K(+) efflux antiporter, and cation/H(+) exchanger families. However, currently, little is known about the CPA genes in Rosaceae species. In this study, 220 CPA genes were identified from five Rosaceae species (Pyrus bretschneideri, Malus domestica, Prunus persica, Fragaria vesca, and Prunus mume), and 53 of which came from P. bretschneideri. Phylogenetic, structure, collinearity, and gene expression analyses were conducted on the entire CPA genes of pear. Gene expression data showed that 35 and 37 CPA genes were expressed in pear fruit and pollen tubes, respectively. The transcript analysis of some CPA genes under abiotic stress conditions revealed that CPAs may play an important role in pollen tubes growth. The results presented here will be useful in improving understanding of the complexity of the CPA gene family and will promote functional characterization in future studies.

  18. Asynchronous ripening behavior of cactus pear (Opuntia ficus-indica) cultivars with respect to physicochemical and physiological attributes.

    Science.gov (United States)

    Kyriacou, M C; Emmanouilidou, M G; Soteriou, G A

    2016-11-15

    Physicochemical and physiological ripening events in cactus pear (Opuntia ficus-indica) fruit of cultivars 'Ntopia' and 'Hercules' were profiled against skin coloration from mature-green (S1) to over-mature (S5). Fructose and glucose accumulation were linear in 'Ntopia' but peaked near S3 in 'Hercules' synchronously to the appearance of sucrose. Betalains increased steadily in 'Ntopia' (103.2mg/l) but peaked before full skin coloration in 'Hercules' (49.7mg/l); whereas phenolic content remained invariable and ascorbate content peaked near S5 in both 'Ntopia' (108.6μg/g) and 'Hercules' (163.1μg/g). Cell wall material diminished with maturity though textural changes with ripening appeared not related to pectin solubilization but to weakening of glycan bonding and loss of neutral sugars. Fruit firmness rather was correlated to seed weight (r=0.89) and seed-to-pulp ratio (r=0.73). Cultivar differences highlighted in the chronology of ripening events are critical for defining optimum harvest maturity and postharvest handling protocols for premium quality cactus pear fruit. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Effect of Salicylic Acid and Chelated Magnesium Sulfate on Fruit Quality Improvement (Physical Characteristics in Pear (cv. Louise Bonne

    Directory of Open Access Journals (Sweden)

    mahjabin adel

    2017-02-01

    Full Text Available Introduction: Fruit quality is described based on the crop functions (for industry or table and/or difference of the consumer tastes in different societies. The conformity of the quality with consumer demands has an effective role in improvement of the marketing process. For example, elongated pears are preferred for the processing industries and conserving productions. The lack of attention to retaining of quality and/or improvement of apparent situation of gardening product in proportion to consumer demands decreases especial consumer acceptance. The necessity of having desired quality characteristics in pear fruits from the characters viewpoint of fruit specific gravity (major rating criterion of pears and proportion of length to diameter (minor rating, because of their role in market acceptance and pricing, is evident. Fruit quality, while harvesting, as one of the components of plants fertility influenced under different parameters like nourishment and could be managed during growth season. In other words, the gain of qualities proportionated to consumers demands and/or processing industries is possible by the use of acquired method such as the kind of mother plant nutrition, and control of pests and diseases, etc. In the current research, the effect of salicylic acid and chelated magnesium sulfate was studied on physical indexes of fruits quality of pear fruit. Materials and Methods: In order to study the effect of treatment agents, an experiment was conducted in the ecological conditions of Qazvinon Pear trees belonging to Louise Bonne cultivar in the Randomized Complete Block Design. The treatments includecontrol group (with andwithout water,chelated magnesium sulfate with concentration of 0.5 gram in a liter, chelated magnesium sulfate with concentration of 0.7gram in a liter, salicylic acid with concentration of0.1 gram in a liter,the compound treatment of salicylic acid with concentration of 0.1 andchelated magnesium sulfate with

  20. Steam explosion treatment for ethanol production from branches pruned from pear trees by simultaneous saccharification and fermentation.

    Science.gov (United States)

    Sasaki, Chizuru; Okumura, Ryosuke; Asada, Chikako; Nakamura, Yoshitoshi

    2014-01-01

    This study investigated the production of ethanol from unutilized branches pruned from pear trees by steam explosion pretreatment. Steam pressures of 25, 35, and 45 atm were applied for 5 min, followed by enzymatic saccharification of the extracted residues with cellulase (Cellic CTec2). High glucose recoveries, of 93.3, 99.7, and 87.1%, of the total sugar derived from the cellulose were obtained from water- and methanol-extracted residues after steam explosion at 25, 35, and 45 tm, respectively. These values corresponded to 34.9, 34.3, and 27.1 g of glucose per 100 g of dry steam-exploded branches. Simultaneous saccharification and fermentation experiments were done on water-extracted residues and water- and methanol-extracted residues by Kluyveromyces marxianus NBRC 1777. An overall highest theoretical ethanol yield of 76% of the total sugar derived from cellulose was achieved when 100 g/L of water- and methanol-washed residues from 35 atm-exploded pear branches was used as substrate.

  1. The ability of a cold-adapted Rhodotorula mucilaginosa strain from Tibet to control blue mold in pear fruit.

    Science.gov (United States)

    Hu, Hao; Yan, Fujie; Wilson, Charles; Shen, Qing; Zheng, Xiaodong

    2015-12-01

    Cold-adapted yeasts were isolated from soil samples collected in Tibet and evaluated as potential biocontrol agents against blue mold (Penicillium expansum) of pear fruit in cold storage. YC1, an isolate identified as Rhodotorula mucilaginosa, was found to exhibit the greatest biocontrol activity among the different isolates that were screened. A washed cell suspension of YC1 exhibited the best biocontrol activity among three different preparations that were used in the current study. A concentration of 10(8) cells/ml reduced the incidence of decay to 35 %, compared to the control where decay incidence was 100 %. A higher intracellular level of trehalose and a higher proportion of polyunsaturated acids present in YC1, was associated with increased the tolerance of this strain to low temperatures, relative to the other strains that were evaluated. The increased tolerance to low temperature allowed the YC1 strain of yeast to more effectively compete for nutrients and space in wounded pear fruit that had been inoculated with spores of P. expansum and placed in cold storage. The present study demonstrated the ability to select cold-adapted yeasts from cold climates and use them as biocontrol agents of postharvest diseases of fruit placed in cold storage.

  2. Unusual immuno-modulatory triterpene-caffeates in the skins of russeted varieties of apples and pears.

    Science.gov (United States)

    Andre, Christelle M; Larsen, Lesley; Burgess, Elaine J; Jensen, Dwayne J; Cooney, Janine M; Evers, Danièle; Zhang, Jingli; Perry, Nigel B; Laing, William A

    2013-03-20

    Three triterpene-caffeates have been isolated from skins of a russeted apple cultivar "Merton Russet" and identified by LC-MS and NMR as betulinic acid-3-cis-caffeate, betulinic acid-3-trans-caffeate, and oleanolic acid-3-trans-caffeate. Betulinic acid-3-trans-caffeate and oleanolic acid-3-trans-caffeate were also found in russeted pear skins. These compounds have not been previously reported in apples or pears, or in any other foods. Their presence was related to suberized tissue as they were only found in russet portions of the partially russeted apple cultivar "Cox's Orange Pippin" and were not detected in the waxy apple cultivar "Royal Gala". High concentrations of betulinic acid-3-trans-caffeate were found in the bark of both "Merton Russet" and "Royal Gala" trees. The three triterpene-caffeates showed anti-inflammatory activity in vitro, inhibiting NF-κB activation with IC50's of 6-9 μM. Betulinic acid-3-trans-caffeate, the predominant compound in the apples, was immuno-modulatory at around 10 μM in the in vitro and ex vivo bioassays, boosting production of the pro-inflammatory cytokine TNFα in cells stimulated with bacterial lipopolysaccharides.

  3. Prodigious polyphyly in imperilled freshwater pearly-mussels (Bivalvia: Unionidae): a phylogenetic test of species and generic designations

    Science.gov (United States)

    Lydeard, Charles; Minton, Russell L.; Williams, James D.

    2000-01-01

    Unionid bivalves or freshwater pearly-mussels (Unionoidea: Unionidae) serve as an exemplary system for examining many of the problems facing systematists and conservation biologists today. Most of the species and genera were described in the late 1800s and early 1900s, but few phylogenetic studies have been conducted to test conventional views of species and classification. Pearly-mussels of Gulf Coastal drainages of the southeastern United States from the Escambia (southern Alabama to Florida) to the Suwannee Rivers (Florida) are a unique fauna comprised of approximately 100 species, with about 30 endemic to the region. In this study, mitochondrial cytochrome c oxidase subunit I and 16S rRNA gene sequences were used to test the monophyly and to estimate evolutionary relationships of five unionid species representing three different genera. The molecular phylogenies depict all three genera as polyphyletic. The prodigious polyphyly exhibited within unionids is due to incorrect notions of homology and false assumptions about missing anatomical data. In contrast, the molecular phylogeny provides evidence to support the recognition of all five unionid species as distinct evolutionary entities. Furthermore, molecular genealogical evidence supports the elevation of Quincuncina infucata (Conrad) of the Suwannee River to species level, for which Q. kleiniana (Lea) is available.

  4. Homogeneous and label-free detection of microRNAs using bifunctional strand displacement amplification-mediated hyperbranched rolling circle amplification.

    Science.gov (United States)

    Zhang, Li-rong; Zhu, Guichi; Zhang, Chun-yang

    2014-07-01

    MicroRNAs (miRNAs) are an emerging class of biomarkers and therapeutic targets for various diseases including cancers. Here, we develop a homogeneous and label-free method for sensitive detection of let-7a miRNA based on bifunctional strand displacement amplification (SDA)-mediated hyperbranched rolling circle amplification (HRCA). The binding of target miRNA with the linear template initiates the bifunctional SDA reaction, generating two different kinds of triggers which can hybridize with the linear template to initiate new rounds of SDA reaction for the production of more and more triggers. In the meantime, the released two different kinds of triggers can function as the first and the second primers, respectively, to initiate the HRCA reaction whose products can be simply monitored by a standard fluorometer with SYBR Green I as the fluorescent indicator. The proposed method exhibits high sensitivity with a detection limit of as low as 1.8 × 10(-13) M and a large dynamic range of 5 orders of magnitude from 0.1 pM to 10 nM, and it can even discriminate the single-base difference among the miRNA family members. Moreover, this method can be used to analyze the total RNA samples from the human lung tissues and might be further applied for sensitive detection of various proteins, small molecules, and metal ions in combination with specific aptamers.

  5. Effectiveness of Azadirachtin (NeemAzal-T/S in Controlling Pear Psylla (Cacopsylla pyri and European Red Mite (Panonychus ulmi

    Directory of Open Access Journals (Sweden)

    Dejan Marčić

    2009-01-01

    Full Text Available Here we present the results of field trials conducted in Serbia to evaluate the effectiveness of a neem-based product, NeemAzal-T/S (containing azadirachtin-A as its active ingredient in the form of an emulsifiable concentrate against pear psylla (Cacopsylla pyri and European red mite (Panonychus ulmi. Efficacy evaluation against C. pyri was carried out in a commercial pear orchard of the Williams pear cultivar, located at Borkovac (Ruma. The insecticides were applied at BBCH 09 pear growth stage, several days before the beginning of hatching of the first generation larvae. The efficacy of azadirachtin was compared to that of mineral oil, abamectin and diflubenzuron. Efficacy evaluation 18 DAT showed total termination of egg laying by C. pyri after treatments with azadirachtin and abamectin, while some new (white eggs were found after treatment with mineral oil. Diflubenzuron treatment failed to fully stop egg laying, but the number of white eggs was significantly lower than it was in the control. Azadirachtin and abamectin achieved 100% efficacy, while the effectiveness of mineral oil was 97.4%, and that of diflubenzuron a mere 59%. All four insecticides significantly reduced the number of older (yellow eggs and larvae, the efficacy being 80.5-92.6% (yellow eggs, 69.8-79.3% (larvae I-III instar and 94.3-100% (larvae IV-V instar. In evaluation 38 DAT, azadirachtin,abamectin and mineral oil achieved 100% efficacy against white and yellow eggs, while diflubenzuron achieved 93% and 86.9% efficacy. All four insecticides were found to demonstrate high efficacy against I-III instar larvae (99.2-100%, but mineral oil treatmentalone achieved high efficacy against IV-V instar larvae (92.4% as well. Efficacy evaluation against P. ulmi was carried out in a commercial orchard of the Red Chief apple cultivar located at Morović (Šid. Azadirachtin efficacy in controlling a summerpopulation of European red mite was compared to a mineral oil, clofentezine and

  6. Three different signal amplification strategies for the impedimetric sandwich detection of thrombin

    Energy Technology Data Exchange (ETDEWEB)

    Ocaña, Cristina; Valle, Manel del, E-mail: manel.delvalle@uab.cat

    2016-03-17

    In this work, we report a comparative study on three highly specific amplification strategies for the ultrasensitive detection of thrombin with the use of aptamer sandwich protocol. The protocol consisted on the use of a first thrombin aptamer immobilized on the electrode surface, the recognition of thrombin protein, and the reaction with a second biotinylated thrombin aptamer forming the sandwich. Through the exposed biotin end, three variants have been tested to amplify the electrochemical impedance signal. The strategies included (a) silver enhancement treatment, (b) gold enhancement treatment and (c) insoluble product produced by the combination of the enzyme horseradish peroxidase (HRP) and 3-amino-9-ethylcarbazole (AEC). The properties of the sensing surface were probed by electrochemical impedance measurements in the presence of the ferrocyanide/ferricyanide redox marker. Insoluble product strategy and silver enhancement treatment resulted in the lowest detection limit (0.3 pM), while gold enhancement method resulted in the highest reproducibility, 8.8% RSD at the pM thrombin concentration levels. Results of silver and gold enhancement treatment also permitted direct inspection by scanning electron microscopy (SEM). - Highlights: • Aptasensor to detect thrombin reaching the femtomolar level. • Biosensing protocol employs two thrombin aptamers in a sandwich capture scheme. • Use of second biotinylated aptamer allows many amplification and detection variants. • Precipitation reaction provides the highest signal amplification of ca. 3 times. • Double recognition event improves remarkably selectivity for thrombin detection.

  7. Peroxy Radical Measurements during the IRRONIC Field Project by C2H6 - NO Chemical Amplification

    Science.gov (United States)

    Wood, E. C. D.; Kundu, S.; Deming, B.; Lew, M.; Stevens, P. S.; Sklaveniti, S.; Dusanter, S.

    2015-12-01

    We present measurements of total peroxy radicals (HO2 + RO2) during the Indiana Radical, Reactivity and Ozone Production Intercomparison (IRRONIC) field project in Bloomington, Indiana during July 2015. Peroxy radicals were measured by chemical amplification using ethane and nitric oxide in dual PFA reaction chambers, and the amplification product NO2 was quantified by cavity attenuated phase shift spectroscopy. On sunny days mid-day peroxy radical mixing ratios were typically between 20 and 70 ppt and were well correlated with "HO2*" measured by the Indiana University Laser-Induced Fluorescence with Fluorescence Assay by Gas Expansion (IU-FAGE) instrument. The ratio of total peroxy radicals (UMass) to the IU-FAGE HO2* measurements was greater than two. We also describe results from an informal intercomparison of the two instruments' calibration sources, which are based on acetone photolysis (UMass) and water photolysis (IU). In addition to sampling the IU calibration source in "amplification" mode, the UMass instrument also separately quantified the HO2 mixing ratio in the IU calibration gas by reaction with excess NO and subsequent quantification of the NO2 produced.

  8. Three different signal amplification strategies for the impedimetric sandwich detection of thrombin

    International Nuclear Information System (INIS)

    Ocaña, Cristina; Valle, Manel del

    2016-01-01

    In this work, we report a comparative study on three highly specific amplification strategies for the ultrasensitive detection of thrombin with the use of aptamer sandwich protocol. The protocol consisted on the use of a first thrombin aptamer immobilized on the electrode surface, the recognition of thrombin protein, and the reaction with a second biotinylated thrombin aptamer forming the sandwich. Through the exposed biotin end, three variants have been tested to amplify the electrochemical impedance signal. The strategies included (a) silver enhancement treatment, (b) gold enhancement treatment and (c) insoluble product produced by the combination of the enzyme horseradish peroxidase (HRP) and 3-amino-9-ethylcarbazole (AEC). The properties of the sensing surface were probed by electrochemical impedance measurements in the presence of the ferrocyanide/ferricyanide redox marker. Insoluble product strategy and silver enhancement treatment resulted in the lowest detection limit (0.3 pM), while gold enhancement method resulted in the highest reproducibility, 8.8% RSD at the pM thrombin concentration levels. Results of silver and gold enhancement treatment also permitted direct inspection by scanning electron microscopy (SEM). - Highlights: • Aptasensor to detect thrombin reaching the femtomolar level. • Biosensing protocol employs two thrombin aptamers in a sandwich capture scheme. • Use of second biotinylated aptamer allows many amplification and detection variants. • Precipitation reaction provides the highest signal amplification of ca. 3 times. • Double recognition event improves remarkably selectivity for thrombin detection.

  9. Social amplification of risk: An empirical study

    International Nuclear Information System (INIS)

    Burns, W.; Slovic, P.; Kasperson, R.; Kasperson, J.; Renn, O.; Emani, S.

    1990-09-01

    The social amplification of risk is a theoretical framework that addresses an important deficiency of formal risk assessment methods and procedures. Typically assessments of risk from technological mishaps have been based upon the expected number of people who could be killed or injured or the amount of property that might be damaged. The diverse and consequential impacts that followed in the aftermath of the Three Mile Island accident make it clear that risk assessments that exclude the role of public perceptions of risk will greatly underestimate the potential costs of certain types of hazards. The accident at Three Mile Island produced no direct fatalities and few, if any, expected deaths due to cancer, yet few other accidents in history have had such costly societal impacts. The experience of amplified impacts argues for the development of a broadened theoretical and methodological perspective capable of integrating technical assessment of risk with public perceptions. This report presents the results to date in an ongoing research effort to better understand the complex processes by which adverse events produce impacts. In particular this research attempts to construct a framework that can account for those events that have produced, or are capable of producing, greater societal impacts than would be forecast by traditional risk assessment methods. This study demonstrates that the social amplification of risk involves interactions between sophisticated technological hazards, public and private institutions, and subtle individual and public perceptions and behaviors. These factors, and the variables underlying the intricate processes of social amplification that occur in modern society, are not fully defined and clarified in this report. 19 refs., 9 figs., 10 tabs

  10. Controlled atmosphere pO2 alters ripening dynamics of 1-MCP treated ‘d’Anjou’ pear (Pyrus communis L.) fruit

    Science.gov (United States)

    Ripening and development of physiological disorders and decay were assessed in ‘d’Anjou’ pear fruit following 1-methylcyclopropene (1-MCP) treatment and cold storage in air or controlled atmosphere (CA). Fruit were exposed after harvest to 0 or 12.6 µmol•L-1 1-MCP and then stored at 0.5 oC in air o...

  11. Pithy brown core in ‘d’Anjou’ pear (Pyrus communis L.) fruit developing during controlled atmosphere storage at pO2 determined by monitoring chlorophyll fluorescense

    Science.gov (United States)

    Physiological responses and fruit quality of ‘d’Anjou’ pear fruit from five orchard lots were evaluated after cold storage in air or controlled atmospheres (CA) with the oxygen (O2) concentration based on assessment of fruit chlorophyll fluorescence (CF) or standard conditions (1.5 kPa O2). The pCO2...

  12. The Relationship between the Morphology and Structure and the Quality of Fruits of Two Pear Cultivars (Pyrus communis L. during Their Development and Maturation

    Directory of Open Access Journals (Sweden)

    Agata Konarska

    2013-01-01

    Full Text Available The flavour and nutritional values of pears are appreciated by consumers worldwide, who, however, demand specific fruit quality, that is, attractive appearance, firmness and flavour, and health safety as well as long-term shelf life and storability. Pear cultivars differ in terms of the above-mentioned traits; therefore, we undertook investigations to demonstrate the differences in structure of fruits of two pear cultivars that determine fruit quality in its broadest sense. The micromorphology, anatomy, and ultrastructure of “Clapp’s Favourite” and “Conference” fruits in the fruit set stage and in the harvest maturity stage were investigated under light microscope and scanning and transmission electron microscopes. The fruits of “Clapp’s Favourite” and “Conference” in the fruit set stage exhibited distinct differences in the values of anatomical parameters only. Substantial differences in fruit structure were observed in the harvest maturity stage. The analyses indicate that firmness and durability of pear fruits are largely influenced by the presence of russeting, the proportion of closed lenticels and number of stone cells, and the content of starch grains and tannin compounds. The thickness of the cuticle and presence of epicuticular waxes as well as the number of lenticels and the number and depth of microcracks play a minor role.

  13. A Quarter Century of Variation in Color and Allometric Characteristics of Eggs from a Rain Forest Population of the Pearly-eyed Thrasher (Margarops fuscatus).

    Science.gov (United States)

    WAYNE J. ARENDT

    2004-01-01

    Egg color, size, and shape vary considerably within and among female Pearly-eyed Thrashers (Margarops fuscatus). Results of a 25-yr study (1979-2004) are presented to provide comparative data. In a sample of 4,128 eggs, typical shape was prolate spheroid; but several variations were observed, depending on the age, stature, and physiological condition of the female, as...

  14. Shell Properties, Water Vapor Loss, and Hatching Success of Eggs from a Rain Forest Population of the Pearly-eyed Thrasher (Margarops fuscatus)

    Science.gov (United States)

    WAYNE J. ARENDT

    2005-01-01

    I calculated various shell properties, water vapor loss, and hatching success of eggs of the Pearly-eyed Thrasher (Margarops fuscatus) using measurements obtained during a long-term study in the Luquillo Mountains, Puerto Rico. Empirical results were comparable to standard reference formulae, demonstrating that published formulae can be used with confidence by field...

  15. Changes in home range of breeding and post-breeding male Pearly-eyed Thrashers in the Luquillo Mountains of Puerto Rico

    Science.gov (United States)

    Jose William Beltran; Joseph M. Wunderle, Jr.; Wayne J. Arendt

    2010-01-01

    Food abundance, time of year, and stage of the reproductive cycle are important factors affecting home range size in birds. Between 23 January and 28 November 2003, we determined the home range and core area sizes for 10 radio-tagged male Pearly-eyed Thrashers (Margarops fuscatus; Mimidae) within the Luquillo Experimental Forest, northeastern Puerto Rico. We found...

  16. Impact of nest predators, competitors, and ectoparasites on Pearly-eyed Thrashers, with comments on the potential implications for Puerto Rican Parrot recovery

    Science.gov (United States)

    Wayne J. Arendt

    2000-01-01

    Over the past 17 years, research on a rain forest population of the Pearly-eyed Thrasher (Margarops fuscatus), with additional observations on nesting Puerto Rican Parrots (Amazona vittata) within the Sierra de Luquillo, Puerto Rico, has shown that reproductive success of thrashers and parrots is often greatly reduced as a result of the additive effects of a diverse...

  17. Characterization of the nutritional components in fruit and cladode of selenium-enriched nutraceutical cactus pear fruit varieties grown on agricultural sediment

    Science.gov (United States)

    Different accessions of different colored cactus pear (Opuntia ficus Indica) were grown in soils high in salts, boron and selenium (Se) located in the Westside of central California. The changes in the nutritional status and biological transformation of the absorbed inorganic Se from the soils into ...

  18. Gas amplification properties of GEM foils

    International Nuclear Information System (INIS)

    Beck, Jeannine

    2009-01-01

    In the framework of the detector concept International Linear Detector for the future accelerator project International Linear Collider, in which electrons and positrons at c. m. energies of 500 GeV are brought to collision, a time projection chamber shall be applied as central track detector. By the application of such a chamber as track detector a three-dimensional reconstruction of the track points is possible. If a particle passes the gas volume within the chamber it ionizises single gas atoms and the arising electrons move after the amplification in the GEM arrangement to the anode, so that a two-dimensional projection of the particle track is possible. The third dimension is calculated from the drift time of the electrons. The advances of this readout system consist therein that a better position resolution than by a multiwire proportional chamber is reached and the back-drifting ions can be strongly suppressed. Aim of this thesis are studies for a GEM module, which shall be used in a large TPC prototype. Concerning different requirements it is valid to compare different GEMs in order to can meet an optimal choice. In a small prototype present at DESY measurements for the acquisition of GEM-describing parameters were performed. The taking into operation of the test TPC was part of this thesis. Tracks were generated by a radioactive source, by means of which the gas amplification was determined. With the measurement arrangement gas-amplifier foils of different kind were compared in view of their amplification properties and their energy resolution power and systematically studied. Five different GEM performances were studied in the test TPC. These foils differ in their geometrical classification parameters, the fabrication process, or the materials. The GEMs produced at CERN possess in comparison with GEMs of the Japanese firm SciEnergy and a GEM of the US-American firm Tech-Etch the best amplification and resolution properties. Furthermore a new GEM framing

  19. Amplification Effects and Unconventional Monetary Policies

    Directory of Open Access Journals (Sweden)

    Cécile BASTIDON GILLES

    2012-02-01

    Full Text Available Global financial crises trigger off amplification effects, which allow relatively small shocks to propagate through the whole financial system. For this reason, the range of Central banks policies is now widening beyond conventional monetary policies and lending of last resort. The aim of this paper is to establish a rule for this practice. The model is based on the formalization of funding conditions in various types of markets. We conduct a comprehensive analysis of the “unconventional monetary policies”, and especially quantify government bonds purchases by the Central bank.

  20. Amplification of curvature perturbations in cyclic cosmology

    International Nuclear Information System (INIS)

    Zhang Jun; Liu Zhiguo; Piao Yunsong

    2010-01-01

    We analytically and numerically show that through the cycles with nonsingular bounce, the amplitude of curvature perturbation on a large scale will be amplified and the power spectrum will redden. In some sense, this amplification will eventually destroy the homogeneity of the background, which will lead to the ultimate end of cycles of the global universe. We argue that for the model with increasing cycles, it might be possible that a fissiparous multiverse will emerge after one or several cycles, in which the cycles will continue only at corresponding local regions.

  1. Parametric Amplification of Gravitational Fluctuations during Reheating

    International Nuclear Information System (INIS)

    Finelli, F.; Brandenberger, R.; Finelli, F.

    1999-01-01

    Cosmological perturbations can undergo amplification by parametric resonance during preheating even on scales larger than the Hubble radius, without violating causality. A unified description of gravitational and matter fluctuations is crucial to determine the strength of the instability. To extract specific signatures of the oscillating inflaton field during reheating, it is essential to focus on a variable describing metric fluctuations which is constant in the standard analyses of inflation. For a massive inflaton without self-coupling, we find no additional growth of superhorizon modes during reheating beyond the usual predictions. For a massless self-coupled inflaton, there is a sub-Hubble scale resonance. copyright 1999 The American Physical Society

  2. Light-Triggered Soft Artificial Muscles: Molecular-Level Amplification of Actuation Control Signals.

    Science.gov (United States)

    Dicker, Michael P M; Baker, Anna B; Iredale, Robert J; Naficy, Sina; Bond, Ian P; Faul, Charl F J; Rossiter, Jonathan M; Spinks, Geoffrey M; Weaver, Paul M

    2017-08-23

    The principle of control signal amplification is found in all actuation systems, from engineered devices through to the operation of biological muscles. However, current engineering approaches require the use of hard and bulky external switches or valves, incompatible with both the properties of emerging soft artificial muscle technology and those of the bioinspired robotic systems they enable. To address this deficiency a biomimetic molecular-level approach is developed that employs light, with its excellent spatial and temporal control properties, to actuate soft, pH-responsive hydrogel artificial muscles. Although this actuation is triggered by light, it is largely powered by the resulting excitation and runaway chemical reaction of a light-sensitive acid autocatalytic solution in which the actuator is immersed. This process produces actuation strains of up to 45% and a three-fold chemical amplification of the controlling light-trigger, realising a new strategy for the creation of highly functional soft actuating systems.

  3. Laser induced nuclear reactions

    International Nuclear Information System (INIS)

    Ledingham, Ken; McCanny, Tom; Graham, Paul; Fang Xiao; Singhal, Ravi; Magill, Joe; Creswell, Alan; Sanderson, David; Allott, Ric; Neely, David; Norreys, Peter; Santala, Marko; Zepf, Matthew; Watts, Ian; Clark, Eugene; Krushelnick, Karl; Tatarakis, Michael; Dangor, Bucker; Machecek, Antonin; Wark, Justin

    1998-01-01

    Dramatic improvements in laser technology since 1984 have revolutionised high power laser technology. Application of chirped-pulse amplification techniques has resulted in laser intensities in excess of 10 19 W/cm 2 . In the mid to late eighties, C. K. Rhodes and K. Boyer discussed the possibility of shining laser light of this intensity onto solid surfaces and to cause nuclear transitions. In particular, irradiation of a uranium target could induce electro- and photofission in the focal region of the laser. In this paper it is shown that μCi of 62 Cu can be generated via the (γ,n) reaction by a laser with an intensity of about 10 19 Wcm -2

  4. Loop-mediated isothermal amplification for rapid and convenient detection of Mycoplasma hyopneumoniae.

    Science.gov (United States)

    Li, Jiahe; Minion, F Chris; Petersen, Andrew C; Jiang, Fei; Yang, Sheng; Guo, Panpan; Li, Jinxiang; Wu, Wenxue

    2013-04-01

    Loop-mediated isothermal amplification (LAMP), a novel method of gene amplification, was employed in this study for detecting Mycoplasma hyopneumoniae in the respiratory tract or lungs of swine. The pathogen can be detected in LAMP reactions containing as few as 10 fg purified target DNA (10 copies of M. hyopneumoniae genome) within 30 min, which was comparable to real-time PCR. After 30-min reaction at 63 °C, the addition of a certain amount of dye (SYBR Green I and hydroxyl naphthol blue at a proper ratio) into the LAMP reaction system makes the results easily determined as positive or negative by visual inspection. In addition, the LAMP was able to distinguish between M. hyopneumoniae and other closely-related mycoplasma strains, indicating a high degree of specificity. The LAMP assay was more simple and cheap, since the reaction could be completed under isothermal conditions and less laboratorial infrastructure are required. And, it was proven reliable for M. hyopneumoniae diagnosis of nasal swab and lung samples from the field.

  5. Nonspecific amplification of human DNA by Streptococcus pneumoniae LytA primer

    Directory of Open Access Journals (Sweden)

    Helen Hencida Thangamony

    2018-01-01

    Full Text Available Background: Determination of various analytical parameters is essential for the validation of primers used for in-house nucleic acid amplification tests. While standardising a high-resolution melt analysis (HRMA for detection of Streptococcus pneumoniae in acute pyogenic meningitis, we encountered non-specific amplification of certain base pair sequences of human DNA by Centers for Disease Control & Prevention, USA recommended S. pneumoniae LytA primer. Materials and Methods: HRMA was standardised using DNA extracted from an ATCC strain of S. pneumoniae using SP LytA F373 primer and Type-it HRMTM polymerase chain reaction kit in Rotor-Gene Q Thermal Cycler according to the manufacturer's instructions. Specificity of the primers was determined in dry and wet laboratory experiments against diverse related and unrelated microbial pathogens by HRMA and on DNA extracted from unspiked clinical samples negative for SP DNA. Sensitivity was determined by calculating lower limit of detection threshold in experiments with spiked samples. The amplicon from spiked experiments was sequenced and analysed through Gene Bank. Results: Our dry/wet laboratory experiments showed two separate curves and different Tm values indicating certain non-specific amplification by the primer. Basic Local Alignment Search Tool (BLAST analysis of the amplicon obtained in the spiked experiment showed sequences of human chromosome 20 associated with Homo sapiens protein tyrosine phosphatase, receptor type T gene. The problem was resolved by stopping the reaction at 30th Ct cycle and observing the Tm values. Conclusion: Since HRMA is done without a specific probe, one should be aware of non-specific amplifications while using primers for HRMA of human clinical samples.

  6. Analytically Sensitive Protein Detection in Microtiter Plates by Proximity Ligation with Rolling Circle Amplification.

    Science.gov (United States)

    Ebai, Tonge; Souza de Oliveira, Felipe Marques; Löf, Liza; Wik, Lotta; Schweiger, Caroline; Larsson, Anders; Keilholtz, Ulrich; Haybaeck, Johannes; Landegren, Ulf; Kamali-Moghaddam, Masood

    2017-09-01

    Detecting proteins at low concentrations in plasma is crucial for early diagnosis. Current techniques in clinical routine, such as sandwich ELISA, provide sensitive protein detection because of a dependence on target recognition by pairs of antibodies, but detection of still lower protein concentrations is often called for. Proximity ligation assay with rolling circle amplification (PLARCA) is a modified proximity ligation assay (PLA) for analytically specific and sensitive protein detection via binding of target proteins by 3 antibodies, and signal amplification via rolling circle amplification (RCA) in microtiter wells, easily adapted to instrumentation in use in hospitals. Proteins captured by immobilized antibodies were detected using a pair of oligonucleotide-conjugated antibodies. Upon target recognition these PLA probes guided oligonucleotide ligation, followed by amplification via RCA of circular DNA strands that formed in the reaction. The RCA products were detected by horseradish peroxidase-labeled oligonucleotides to generate colorimetric reaction products with readout in an absorbance microplate reader. We compared detection of interleukin (IL)-4, IL-6, IL-8, p53, and growth differentiation factor 15 (GDF-15) by PLARCA and conventional sandwich ELISA or immuno-RCA. PLARCA detected lower concentrations of proteins and exhibited a broader dynamic range compared to ELISA and iRCA using the same antibodies. IL-4 and IL-6 were detected in clinical samples at femtomolar concentrations, considerably lower than for ELISA. PLARCA offers detection of lower protein levels and increased dynamic ranges compared to ELISA. The PLARCA procedure may be adapted to routine instrumentation available in hospitals and research laboratories. © 2017 American Association for Clinical Chemistry.

  7. Different biosynthesis patterns among flavonoid 3-glycosides with distinct effects on accumulation of other flavonoid metabolites in pears (Pyrus bretschneideri Rehd..

    Directory of Open Access Journals (Sweden)

    Rui Zhai

    Full Text Available Flavonoid biosynthesis profile was clarified by fruit bagging and re-exposure treatments in the green Chinese pear 'Zaosu' (Pyrus bretschneideri Rehd. and its red mutant 'Red Zaosu'. Two distinct biosynthesis patterns of flavonoid 3-glycosides were found in 'Zaosu' pear. By comparison with 'Red Zaosu', the biosynthesis of flavonoid 3-galactosides and flavonoid 3-arabinosides were inhibited by bagging and these compounds only re-accumulated to a small degree in the fruit peel of 'Zaosu' after the bags were removed. In contrast, the biosynthesis of flavonoid 3-gluctosides and flavonoid 3-rutinosides was reduced by bagging and then increased when the fruits were re-exposed to sunlight. A combination of correlation, multicollinearity test and partial-correlation analyses among major flavonoid metabolites indicated that biosynthesis of each phenolic compound was independent in 'Zaosu' pear, except for the positive correlation between flavonoid 3-rutincosides and flavanols. In contrast with the green pear cultivar, almost all phenolic compounds in the red mutant had similar biosynthesis patterns except for arbutin. However, only the biosynthesis of flavonoid 3-galactosides was relatively independent and strongly affected the synthesis of the other phenolic compounds. Therefore, we propose a hypothesis that the strong accumulation of flavonoid 3-galactosides stimulated the biosynthesis of other flavonoid compounds in the red mutant and, therefore, caused systemic variation of flavonoid biosynthesis profiles between 'Zaosu' and its red mutant. This hypothesis had been further demonstrated by the enzyme activity of UFGT, and transcript levels of flavonoid biosynthetic genes and been well tested by a stepwise linear regression forecasting model. The gene that encodes flavonoid 3-galacosyltransferase was also identified and isolated from the pear genome.

  8. C-MET overexpression and amplification in gliomas.

    Science.gov (United States)

    Kwak, Yoonjin; Kim, Seong-Ik; Park, Chul-Kee; Paek, Sun Ha; Lee, Soon-Tae; Park, Sung-Hye

    2015-01-01

    We investigated c-Met overexpression and MET gene amplification in gliomas to determine their incidence and prognostic significance. c-Met immunohistochemistry and MET gene fluorescence in situ hybridization were carried out on tissue microarrays from 250 patients with gliomas (137 grade IV GBMs and 113 grade II and III diffuse gliomas). Clinicopathological features of these cases were reviewed. c-Met overexpression and MET gene amplification were detected in 13.1% and 5.1% of the GBMs, respectively. All the MET-amplified cases showed c-Met overexpression, but MET amplification was not always concordant with c-Met overexpression. None of grade II and III gliomas demonstrated c-Met overexpression or MET gene amplification. Mean survival of the GBM patients with MET amplification was not significantly different from patients without MET amplification (P=0.155). However, GBM patients with c-Met overexpression survived longer than patients without c-Met overexpression (P=0.035). Although MET amplification was not related to poor GBM prognosis, it is partially associated with the aggressiveness of gliomas, as MET amplification was found only in grade IV, not in grade II and III gliomas. We suggest that MET inhibitor therapy may be beneficial in about 5% GBMs, which was the incidence of MET gene amplification found in the patients included in this study.

  9. Earthquake acceleration amplification based on single microtremor test

    Science.gov (United States)

    Jaya Syahbana, Arifan; Kurniawan, Rahmat; Soebowo, Eko

    2018-02-01

    Understanding soil dynamics is needed to understand soil behaviour, including the parameters of earthquake acceleration amplification. Many researchers now conduct single microtremor tests to obtain amplification of velocity and natural periods of soil at test sites. However, these amplification parameters are rarely used, so a method is needed to convert the velocity amplification to acceleration amplification. This paper will discuss the proposed process of changing the value of amplification. The proposed method is to integrate the time histories of the synthetic earthquake acceleration of the soil surface under the deaggregation at that location so the time histories of the velocity earthquake will be obtained. Next is to conduct a “fitting curve” between amplification by a single microtremor test with amplification of the synthetic earthquake velocity time histories. After obtaining the fitting curve time histories of velocity, differentiation will be conducted to obtain fitting curve acceleration time histories. The final step after obtaining the fitting curve is to compare the acceleration of the “fitting curve” against the histories time of the acceleration of synthetic earthquake at bedrocks to obtain single microtremor acceleration amplification factor.

  10. Quantum tomography enhanced through parametric amplification

    Science.gov (United States)

    Knyazev, E.; Spasibko, K. Yu; Chekhova, M. V.; Khalili, F. Ya

    2018-01-01

    Quantum tomography is the standard method of reconstructing the Wigner function of quantum states of light by means of balanced homodyne detection. The reconstruction quality strongly depends on the photodetectors quantum efficiency and other losses in the measurement setup. In this article we analyze in detail a protocol of enhanced quantum tomography, proposed by Leonhardt and Paul [1] which allows one to reduce the degrading effect of detection losses. It is based on phase-sensitive parametric amplification, with the phase of the amplified quadrature being scanned synchronously with the local oscillator phase. Although with sufficiently strong amplification the protocol enables overcoming any detection inefficiency, it was so far not implemented in the experiment, probably due to the losses in the amplifier. Here we discuss a possible proof-of-principle experiment with a traveling-wave parametric amplifier. We show that with the state-of-the-art optical elements, the protocol enables high fidelity tomographic reconstruction of bright non-classical states of light. We consider two examples: bright squeezed vacuum and squeezed single-photon state, with the latter being a non-Gaussian state and both strongly affected by the losses.

  11. Electronic cyclotron radiation amplification in thermonuclear plasmas

    International Nuclear Information System (INIS)

    Ziebell, L.F.

    1983-01-01

    The amplified emission of electron cyclotron radiation near the fundamental frequency from an inhomogeneous, anisotropic plasma slab is investigated in a linear theory. Plasma polarization effects are consistently included. Expressions are developed in the WKB approximation for emission in the ordinary and the extraordinary modes, for propagation perpendicular to the magnetic field. Numerical results are given for the extraordinary mode, for which effects are strongest. For the case of a loss-cone-type electron momentum distribution, it is shown that the amplification is sensitively dependent on the ratio of parallel-to-perpendicular temperature and on inhomogeneities in the magnetic field. The dependence of the amplification on the distribution is further investigated by considering superpositions of loss-cone and Maxwellian components. It is show that the presence of a Maxwellian component in general reduces the emission relative to the pure loss-cone case, and situations occur in which a layer in the slab very effectively absorbs all the radiation amplified elsewhere. A peculiar behaviour of the refractive index, which occurs in the transition from the pure loss-cone to the pure Maxwellian case, is discussed. (author)

  12. Protein enrichment of cactus pear (Opuntia ficus - indica Mill using Saccharomyces cerevisiae in solid-state fermentation

    Directory of Open Access Journals (Sweden)

    Lúcia de Fátima Araújo

    2005-06-01

    Full Text Available The microbial protein bioconversion of cactus pear by yeast in solid medium was studied. Three cultivation variables used were: inoculum's concentrations (5, 10 and 15 %, substrate layer thickness (2, 4 and 6 cm and temperature (30, 34 and 38 ºC. The rate of dry matter production and total protein were determined. Results obtained were variance analysis, gross energy and in vitro dry matter digestibility. The maximum protein amount achieved for the conditions studied in the present work was higher than 26 %, which was compatible or greater than those of conventional concentrates of protein supplements used for animal feed. The protein concentrate of cactus pear had a higher in vitro digestibility index (95.8 % and did not show any changes in the gross energy value when compared to that of the cactus pear in natura.A bioconversão da proteína microbiana através da levedura em meio sólido, foi estudada em palma forrageira cultivada em condições laboratoriais, sob três níveis de concentração do inóculo (5, 10 e 15%, espessuras distintas das camadas dos substratos (2, 4 e 6cm e temperaturas (30, 34 e 38ºC. Foram analisadas as taxas de produção de matéria seca (MS, proteína bruta (PB, cujos resultados foram submetidos à análise de variância, energia bruta (EB e digestibilidade in vitro da matéria seca (DIVMS. O valor máximo de teor protéico, alcançado nas condições estudadas nesse trabalho, foi superior a 26%, sendo esse teor compatível ou maior do que os concentrados convencionais utilizados como suplemento protéico para a ração animal. O concentrado protéico da palma obteve um alto índice de digestibilidade in vitro (95,8% e não apresentou grande alteração no valor da energia bruta se comparada com a palma in natura.

  13. Combined effect of temperature and controlled atmosphere on storage and shelf-life of 'Rocha' pear treated with 1-methylcyclopropene.

    Science.gov (United States)

    Gago, Custódia M L; Miguel, Maria G; Cavaco, Ana M; Almeida, Domingos P F; Antunes, Maria D C

    2015-03-01

    The combination of temperature and atmosphere composition for storage of Pyrus communis L. 'Rocha' treated with 1-methylcyclopropene was investigated. Fruits treated with 312 nl l(-1) 1-methylcyclopropene were stored at 0 ℃ and 2.5 ℃ in air and controlled atmosphere (CA) (3.04 kPa O2+ 0.91 kPa CO2). Fruits were removed from storage after 14, 26 and 35 weeks, transferred to shelf-life at approximately 22 ℃ and assessed for ripening and quality, symptoms of superficial scald and internal browning and the accumulation of biochemical compounds related to scald after 0, 1 and 2 weeks. Superficial scald occurred only in fruits stored for 35 weeks in air at 2.5 ℃. Levels of conjugated trienols and α-farnesene increased during the first 26 weeks in storage, remaining constant thereafter. During shelf-life, conjugated trienols were higher in fruits stored in air at 2.5 ℃. Internal browning developed in shelf-life after 26 weeks at 2.5 ℃. Pears in air at 2.5 ℃ were not able to stand a 2-week shelf-life after 35 weeks of storage, while fruits stored at 0 ℃ under CA ripened slowly after the same storage period. The retention of firmness during shelf-life of 1-methylcyclopropene-treated 'Rocha' pear can be overcome by elevating the storage temperature from 0 ℃ to 2.5 ℃, but CA is a required complement to avoid excessive softening after long-term storage. The ratio carotenoid/chlorophyll increased during storage and shelf-life, as plastids senesced. CA reduced the rate of chlorophyll loss during the first 14 weeks in storage, but its effect was reduced afterwards. 'Rocha' pear treated with 1-methylcyclopropene had a similar post-harvest behaviour during long-term storage at 0 ℃ in air or at 2.5 ℃ under CA. © The Author(s) 2013 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  14. Targeted deep resequencing identifies coding variants in the PEAR1 gene that play a role in platelet aggregation.

    Directory of Open Access Journals (Sweden)

    Yoonhee Kim

    Full Text Available Platelet aggregation is heritable, and genome-wide association studies have detected strong associations with a common intronic variant of the platelet endothelial aggregation receptor1 (PEAR1 gene both in African American and European American individuals. In this study, we used a sequencing approach to identify additional exonic variants in PEAR1 that may also determine variability in platelet aggregation in the GeneSTAR Study. A 0.3 Mb targeted region on chromosome 1q23.1 including the entire PEAR1 gene was Sanger sequenced in 104 subjects (45% male, 49% African American, age = 52±13 selected on the basis of hyper- and hypo- aggregation across three different agonists (collagen, epinephrine, and adenosine diphosphate. Single-variant and multi-variant burden tests for association were performed. Of the 235 variants identified through sequencing, 61 were novel, and three of these were missense variants. More rare variants (MAF<5% were noted in African Americans compared to European Americans (108 vs. 45. The common intronic GWAS-identified variant (rs12041331 demonstrated the most significant association signal in African Americans (p = 4.020×10(-4; no association was seen for additional exonic variants in this group. In contrast, multi-variant burden tests indicated that exonic variants play a more significant role in European Americans (p = 0.0099 for the collective coding variants compared to p = 0.0565 for intronic variant rs12041331. Imputation of the individual exonic variants in the rest of the GeneSTAR European American cohort (N = 1,965 supports the results noted in the sequenced discovery sample: p = 3.56×10(-4, 2.27×10(-7, 5.20×10(-5 for coding synonymous variant rs56260937 and collagen, epinephrine and adenosine diphosphate induced platelet aggregation, respectively. Sequencing approaches confirm that a common intronic variant has the strongest association with platelet aggregation in African Americans

  15. Review:Whole genome amplification in preimplantation genetic diagnosis

    Institute of Scientific and Technical Information of China (English)

    Ying-ming ZHENG; Ning WANG; Lei LI; Fan JIN

    2011-01-01

    Preimplantation genetic diagnosis(PGD)refers to a procedure for genetically analyzing embryos prior to implantation,improving the chance of conception for patients at high risk of transmitting specific inherited disorders.This method has been widely used for a large number of genetic disorders since the first successful application in the early 1990s.Polymerase chain reaction(PCR)and fluorescent in situ hybridization(FISH)are the two main methods in PGD,but there are some inevitable shortcomings limiting the scope of genetic diagnosis.Fortunately,different whole genome amplification(WGA)techniques have been developed to overcome these problems.Sufficient DNA can be amplified and multiple tasks which need abundant DNA can be performed.Moreover,WGA products can be analyzed as a template for multi-loci and multi-gene during the subsequent DNA analysis.In this review,we will focus on the currently available WGA techniques and their applications,as well as the new technical trends from WGA products.

  16. A new ultrasonic signal amplification method for detection of bacteria

    Science.gov (United States)

    Kant Shukla, Shiva; Resa López, Pablo; Sierra Sánchez, Carlos; Urréjola, José; Segura, Luis Elvira

    2012-10-01

    A new method is presented that increases the sensitivity of ultrasound-based techniques for detection of bacteria. The technique was developed for the detection of catalase-positive microorganisms. It uses a bubble trapping medium containing hydrogen peroxide that is mixed with the sample for microbiological evaluation. The enzyme catalase is present in catalase-positive bacteria, which induces a rapid hydrolysis of hydrogen peroxide, forming bubbles which remain in the medium. This reaction results in the amplification of the mechanical changes that the microorganisms produce in the medium. The effect can be detected by means of ultrasonic wave amplitude continuous measurement since the bubbles increase the ultrasonic attenuation significantly. It is shown that microorganism concentrations of the order of 105 cells ml-1 can be detected using this method. This allows an improvement of three orders of magnitude in the ultrasonic detection threshold of microorganisms in conventional culture media, and is competitive with modern rapid microbiological methods. It can also be used for the characterization of the enzymatic activity.

  17. A new ultrasonic signal amplification method for detection of bacteria

    International Nuclear Information System (INIS)

    Shukla, Shiva Kant; López, Pablo Resa; Sánchez, Carlos Sierra; Segura, Luis Elvira; Urréjola, José

    2012-01-01

    A new method is presented that increases the sensitivity of ultrasound-based techniques for detection of bacteria. The technique was developed for the detection of catalase-positive microorganisms. It uses a bubble trapping medium containing hydrogen peroxide that is mixed with the sample for microbiological evaluation. The enzyme catalase is present in catalase-positive bacteria, which induces a rapid hydrolysis of hydrogen peroxide, forming bubbles which remain in the medium. This reaction results in the amplification of the mechanical changes that the microorganisms produce in the medium. The effect can be detected by means of ultrasonic wave amplitude continuous measurement since the bubbles increase the ultrasonic attenuation significantly. It is shown that microorganism concentrations of the order of 10 5 cells ml −1 can be detected using this method. This allows an improvement of three orders of magnitude in the ultrasonic detection threshold of microorganisms in conventional culture media, and is competitive with modern rapid microbiological methods. It can also be used for the characterization of the enzymatic activity. (paper)

  18. Risks, media and the social amplification of soil contamination

    Energy Technology Data Exchange (ETDEWEB)

    Ouboter, S. [NOK, Networkorganisation for Environmental Quality, Gouda (Netherlands)

    2003-07-01

    Soil experts think of the risks of contaminated sites in terms of adverse effects of toxic substances on human health or environmental quality. In other words, the risk is attributed to the contamination. Social scientists define risk as a situation or event in which something of human value (including humans themselves) has been put at stake and where the outcome is uncertain. Since situations or events are constructions of the human mind, risks are also constructed. A relevant question for a psychologist is to learn how these constructions evolve in the mind of an individual and how this perceived risk influences the individuals' behaviour and well-being. A relevant question for a sociologist is how individuals with their own perceptions, feelings and behaviour interact. Many soil contamination experts experienced that one a site is seen as contaminated by a loathsome source, a chain of adverse reactions can easily put a stigma on that specific location and groups of people associated with that contaminated site. The case of Love Canal is worldwide known as an example of this phenomenon, but many countries have their own national symbol, like Lekkerkerk in the Netherlands. Modern media play an important role in this process. This process is often believed to be irrational and therefore uncontrollable. The question of this workshop is to what level technical soil experts can influence the psychological and social effects of soil contamination, using the social amplification metaphor. (orig.)

  19. Ensacamento de frutos de pereira cv. Housui Bagging of nashi pear cv. Housui

    Directory of Open Access Journals (Sweden)

    Ivan Dagoberto Faoro

    2004-04-01

    Full Text Available Foi avaliada a qualidade de frutos de pêra japonesa cv. Housui (Pyrus pyrifolia var. culta ensacados com diferentes tipos de sacos de papel e em duas épocas: 34 e 83 dias após a florada. O ensacamento não influenciou na firmeza, no teor de sólidos solúveis totais e no peso médio dos frutos. Sacos vermelhos não induziram aumento do pH da polpa, mas o uso de sacos duplos, sacos marrons, sacos de papel kraft marrons e sacos de pipoca brancos aumentaram significativamente o pH. Ao buscar-se maior precocidade de colheita, os melhores resultados foram obtidos com o uso de sacos pequenos de papel manteiga aos 34 dias após a floração e o uso de sacos grandes duplos ou sacos grandes marrons, 83 dias após a plena floração. O ensacamento 34 dias após a plena floração, com sacos grandes de papel duplo de cor marrom ou sacos de papel kraft marrons, ou ainda o uso de sacos pequenos parafinados transparentes de papel manteiga, aos 34 dias, seguidos pela colocação, aos 83 dias, dos dois tipos de sacos grandes citados anteriormente, resultaram em frutos de melhor qualidade externa (película de coloração homogênea e mais clara, lisa e com lenticelas pouco salientes. O uso de sacos vermelhos de papel manteiga e de sacos de pipoca brancos, com ou sem ensacamento prévio com sacos pequenos de papel manteiga parafinado, não resultaram em melhoria substancial da qualidade externa do fruto.The fruit quality of Nashi pears cv. Housui (Pyrus pyrifolia var. culta have been evaluated after bagging with different paper bags at two bagging dates: 34 and 83 days after full bloom. The bagging did not affect the fruit firmness, the total soluble solids and the average fruit weight. Red bags did not affect the pH of the fruit flesh. However the fruit bagged with double bags of brown color, brown kraft paper and white popcorn paper increased the pH. In order to harvest precociously, the use of small transparent paraffin paper bags 34 days after full bloom

  20. Phytochemicals, nutritionals and antioxidant properties of two prickly pear cactus cultivars (Opuntia ficus indica Mill.) growing in Taif, KSA.

    Science.gov (United States)

    Abdel-Hameed, El-Sayed S; Nagaty, Mohamed A; Salman, Mahmood S; Bazaid, Salih A

    2014-10-01

    The antioxidant properties, some phytochemicals and nutritionals were characterized in two prickly pear cactus (Opuntia ficus indica Mill.) cultivars; red and yellow; growing in Taif, Kingdom of Saudi Arabia (KSA). The antioxidant properties of red cactus cultivar were higher than the yellow cactus cultivar. Linear correlation appeared between the antioxidant properties and total phenolics. All samples nearly have the same quantity of iron, copper, sodium and potassium. Some phenolic compounds were detected by HPLC-UV analysis. HPLC-RI analysis of all samples revealed the absence of sucrose and the presence of glucose and fructose. According to the above results, this study gave a good indication about the nutritional and pharmaceutical potential of the two cactus cultivars that must be widespread cultivated in arid and semiarid regions as KSA accompanying with establishment of industries beside the cactus farms that used all parts of plants. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Variability of some morphological traits of one-year old red oak, black walnut, birch and wild pear seedlings in the nurseries of Jastrebac region

    Directory of Open Access Journals (Sweden)

    Krstić Milun

    2002-01-01

    Full Text Available Five morphological and quantitative characteristics of one-year old seedlings of Red oak (Quercus rubra L, Black walnut (Juglans nigra L, Wild pear (Pyrus pygrowser Borkh and Birch (Betula verrucosa Ehrh were studied. The seedlings were produced and cultivated in the controlled conditions of the nursery in the region of Jastrebac, by the classical method. Aboveground seedling height, root collar diameter, root length, number of secondary roots and the leaf assimilation area were analysed. Intraspecific and interspecific variability of morphological features of the above species were assessed by the comparative analysis and statistical methods The comparative analysis shows the great individual variability of seedlings, which can indicate their genetic potential, adaptation to environment conditions, further spontaneous selection and the development in natural conditions. This justifies the need of the quality assessment and the first selection already in the nursery, in order to ensure the quality planting material and to reduce the risk of afforestation failure One-year old birch seedlings have the lowest average height (18.8 cm. Black walnut and Wild pear seedlings are approximately twice as high, and Red oak about 2.5 times higher. At the same time Red oak seedlings have for about one-fourth greater height than Black walnut, and for one-third greater height than Wild pear. Wild pear seedlings attain the averagely twice larger root collar diameter than Birch (2.8 cm, Red oak seedlings about 2.5 times larger diameter, and Black walnut 3.5 times larger diameter. Black walnut has a larger root collar diameter than Red oak for about one third, and almost twice larger than Wild pear. Birch, Red oak and Wild pear have almost twice longer root (1.8-1.9 times, Black walnut about 2.25 times longer. The total assimilation area of a Birch seedling is averagely 89.0 cm2. Compared to birch, wild pear has approximately double assimilation area per tree, Red

  2. Genome-Wide Identification, Evolution and Functional Divergence of MYB Transcription Factors in Chinese White Pear (Pyrus bretschneideri).

    Science.gov (United States)

    Li, Xiaolong; Xue, Cheng; Li, Jiaming; Qiao, Xin; Li, Leiting; Yu, Li'ang; Huang, Yuhua; Wu, Jun

    2016-04-01

    The MYB superfamily is large and functionally diverse in plants. To date, MYB family genes have not yet been identified in Chinese white pear (Pyrus bretschneideri), and their functions remain unclear. In this study, we identified 231 genes as candidate MYB genes and divided them into four subfamilies. The R2R3-MYB (PbrMYB) family shared an R2R3 domain with 104 amino acid residues, including five conserved tryptophan residues. The Pbr MYB family was divided into 37 functional subgroups including 33 subgroups which contained both MYB genes of Rosaceae plants and AtMYB genes, and four subgroups which included only Rosaceae MYB genes or AtMYB genes. PbrMYB genes with similar functions clustered into the same subgroup, indicating functional conservation. We also found that whole-genome duplication (WGD) and dispersed duplications played critical roles in the expansion of the MYB family. The 87 Pbr MYB duplicated gene pairs dated back to the two WGD events. Purifying selection was the primary force driving Pbr MYB gene evolution. The 15 gene pairs presented 1-7 codon sites under positive selection. A total of 147 expressed genes were identified from RNA-sequencing data of fruit, and six Pbr MYB members in subgroup C1 were identified as important candidate genes in the regulation of lignin synthesis by quantitative real-time PCR analysis. Further correlation analysis revealed that six PbrMYBs were significantly correlated with five structural gene families (F5H, HCT, CCR, POD and C3'H) in the lignin pathway. The phylogenetic, evolution and expression analyses of the MYB gene family in Chinese white pear establish a solid foundation for future comprehensive functional analysis of Pbr MYB genes. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  3. Emission-Line Galaxies from the PEARS Hubble Ultra Deep Field: A 2-D Detection Method and First Results

    Science.gov (United States)

    Gardner, J. P.; Straughn, Amber N.; Meurer, Gerhardt R.; Pirzkal, Norbert; Cohen, Seth H.; Malhotra, Sangeeta; Rhoads, james; Windhorst, Rogier A.; Gardner, Jonathan P.; Hathi, Nimish P.; hide

    2007-01-01

    The Hubble Space Telescope (HST) Advanced Camera for Surveys (ACS) grism PEARS (Probing Evolution And Reionization Spectroscopically) survey provides a large dataset of low-resolution spectra from thousands of galaxies in the GOODS North and South fields. One important subset of objects in these data are emission-line galaxies (ELGs), and we have investigated several different methods aimed at systematically selecting these galaxies. Here we present a new methodology and results of a search for these ELGs in the PEARS observations of the Hubble Ultra Deep Field (HUDF) using a 2D detection method that utilizes the observation that many emission lines originate from clumpy knots within galaxies. This 2D line-finding method proves to be useful in detecting emission lines from compact knots within galaxies that might not otherwise be detected using more traditional 1D line-finding techniques. We find in total 96 emission lines in the HUDF, originating from 81 distinct "knots" within 63 individual galaxies. We find in general that [0 1111 emitters are the most common, comprising 44% of the sample, and on average have high equivalent widths (70% of [0 1111 emitters having rest-frame EW> 100A). There are 12 galaxies with multiple emitting knots; several show evidence of variations in H-alpha flux in the knots, suggesting that the differing star formation properties across a single galaxy can in general be probed at redshifts approximately greater than 0.2 - 0.4. The most prevalent morphologies are large face-on spirals and clumpy interacting systems, many being unique detections owing to the 2D method described here, thus highlighting the strength of this technique.

  4. The rolling circle amplification and next generation sequencing ...

    African Journals Online (AJOL)

    Rolling circle amplification is a simple approach of enriching populations of single-stranded DNA plant begomovirus genomes (genus, Begomovirus; family, Geminiviridae). This is an innovative approach that utilizes the robustness of the bacteriophage phi29 DNA polymerase used in circle amplification, together with deep ...

  5. Amplification of Chirality in Hydrogen-Bonded Tetrarosette Helices

    NARCIS (Netherlands)

    Mateos timoneda, Miguel; Crego Calama, Mercedes; Reinhoudt, David

    2006-01-01

    The amplification of chirality in hydrogen-bonded tetrarosette assemblies under thermodynamic equilibrium is described. The extent of the chiral amplification obtained by means of “sergeants-and-soldiers” experiments depends only on the structure of the assembly and it is independent of the

  6. Centrosome Amplification Is Sufficient to Promote Spontaneous Tumorigenesis in Mammals

    NARCIS (Netherlands)

    Levine, Michelle S.; Bakker, Bjorn; Boeckx, Bram; Moyett, Julia; Lu, James; Vitre, Benjamin; Spierings, Diana C.; Lansdorp, Peter M.; Cleveland, Don W.; Lambrechts, Diether; Foijer, Floris; Holland, Andrew J.

    2017-01-01

    Centrosome amplification is a common feature of human tumors, but whether this is a cause or a consequence of cancer remains unclear. Here, we test the consequence of centrosome amplification by creating mice in which centrosome number can be chronically increased in the absence of additional

  7. Explanatory Model for Sound Amplification in a Stethoscope

    Science.gov (United States)

    Eshach, H.; Volfson, A.

    2015-01-01

    In the present paper we suggest an original physical explanatory model that explains the mechanism of the sound amplification process in a stethoscope. We discuss the amplification of a single pulse, a continuous wave of certain frequency, and finally we address the resonant frequencies. It is our belief that this model may provide students with…

  8. Chirped pulse amplification: Present and future

    International Nuclear Information System (INIS)

    Maine, P.; Strickland, D.; Pessot, M.; Squier, J.; Bado, P.; Mourou, G.; Harter, D.

    1988-01-01

    Short pulses with ultrahigh peak powers have been generated in Nd: glass and Alexandrite using the Chirped Pulse Amplification (CPA) technique. This technique has been successful in producing picosecond terawatt pulses with a table-top laser system. In the near future, CPA will be applied to large laser systems such as NOVA to produce petawatt pulses (1 kJ in a 1 ps pulse) with focused intensities exceeding 10/sup /plus/21/ W/cm 2 . These pulses will be associated with electric fields in excess of 100 e/a/sub o/ 2 and blackbody energy densities equivalent to 3 /times/ 10 10 J/cm 3 . This petawatt source will have important applications in x-ray laser research and will lead to fundamentally new experiments in atomic, nuclear, solid-state, plasma, and high-energy density physics. A review of present and future designs are discussed. 17 refs., 5 figs

  9. Raman amplification in optical communication systems

    DEFF Research Database (Denmark)

    Kjær, Rasmus

    2008-01-01

    Fiber Raman amplifiers are investigated with the purpose of identifying new applications and limitations for their use in optical communication systems. Three main topics are investigated, namely: New applications of dispersion compensating Raman amplifiers, the use Raman amplification to increase...... fiberbaserede Raman-forstærkere med henblik på at identificere både deres begrænsninger og nye anvendelsesmuligheder i optiske kommunikationssystemer. En numerisk forstærkermodel er blevet udviklet for bedre at forstå forstærkerens dynamik, dens gain- og støjbegrænsninger. Modellen bruges til at forudsige...... forstærkerens statiske og dynamiske egenskaber, og det eftervises at dens resultater er i god overensstemmelse med eksperimentelle forstærkermålinger. Dispersions-kompenserende fiber er på grund af sin store udbredelse og fiberens høje Raman gain effektivitet et meget velegnet Raman gain-medium. Tre nye...

  10. Pain Amplification Syndrome: A Biopsychosocial Approach.

    Science.gov (United States)

    Namerow, Lisa B; Kutner, Emily C; Wakefield, Emily C; Rzepski, Barbara R; Sahl, Robert A

    2016-08-01

    Pediatric neurologists frequently encounter patients who present with significant musculoskeletal pain that cannot be attributed to a specific injury or illness, which can often be defined as pain amplification syndrome (PAS). PAS in children and adolescents is the result of a heightened pain sensitivity pathway, which is intensified by significant biological, psychological, and social contributors. Appropriate assessment and multimodal intervention of PAS are crucial to treatment success, including neurology and behavioral health collaborative treatment plans to restore patient function and reduce pain perception. Pediatric neurologists are imperative in the identification of patients with PAS, providing the family assurance in diagnosis and validation of pain, and directing patients to the appropriate multidisciplinary treatment pathway. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Social and political amplification of technological hazards

    International Nuclear Information System (INIS)

    Ibitayo, Olurominiyi O.; Mushkatel, Alvin; Pijawka, K. David

    2004-01-01

    Using an industrial explosion in Henderson, Nevada, as a case study, this paper examines three main issues: the efficacy of a technological hazard event in amplifying otherwise latent issues, the extent to which the hazard event can serve as a focusing event for substantive local and state policy initiatives, and the effect of fragmentation of political authority in managing technological hazards. The findings indicate that the explosion amplified several public safety issues and galvanized the public into pressing for major policy initiatives. However, notwithstanding the amplification of several otherwise latent issues, and the flurry of activities by the state and local governments, the hazard event did not seem to be an effective focusing event or trigger mechanism for substantive state and local policy initiatives. In addition, the study provides evidence of the need for a stronger nexus between political authority, land-use planning and technological hazard management

  12. Resistive wall modes and error field amplification

    International Nuclear Information System (INIS)

    Boozer, Allen H.

    2003-01-01

    Resistive wall modes and the rapid damping of plasma rotation by the amplification of magnetic field errors are related physical phenomena that affect the performance of the advanced tokamak and spherical torus plasma confinement devices. Elements of our understanding of these phenomena and the code that is used to design the major experimental facilities are based on the electrical circuit representation of the response of the plasma to perturbations. Although the circuit representation of the plasma may seem heuristic, this representation can be rigorously obtained using Maxwell's equations and linearity for plasmas that evolve on a disparate time scale from that of external currents. These and related results are derived. In addition methods are given for finding the plasma information that the circuit representation requires using post-processors for codes that calculate perturbed plasma equilibria

  13. Scintillation light detectors with Neganov Luke amplification

    Science.gov (United States)

    Isaila, C.; Boslau, O.; Coppi, C.; Feilitzsch, F. v.; Goldstraß, P.; Jagemann, T.; Jochum, J.; Kemmer, J.; Lachenmaier, T.; Lanfranchi, J.-C.; Pahlke, A.; Potzel, W.; Rau, W.; Stark, M.; Wernicke, D.; Westphal, W.

    2006-04-01

    For an active suppression of the gamma and electron background in the Cryogenic Rare Event Search with Superconducting Thermometers (CRESST) dark matter experiment both phonons and scintillation light generated in a CaWO 4 crystal are detected simultaneously. The phonon signal is read out by a transition edge sensor (TES) on the CaWO 4 crystal. For light detection a silicon absorber equipped with a TES is employed. An efficient background discrimination requires very sensitive light detectors. The threshold can be improved by applying an electric field to the silicon crystal leading to an amplification of the thermal signal due to the Neganov-Luke effect. Measurements showing the improved sensitivity of the light detectors as well as future steps for reducing the observed extra noise will be presented.

  14. Raman Amplification with a Flying Focus

    Science.gov (United States)

    Turnbull, D.; Bucht, S.; Davies, A.; Haberberger, D.; Kessler, T.; Shaw, J. L.; Froula, D. H.

    2018-01-01

    We propose a new laser amplifier scheme utilizing stimulated Raman scattering in plasma in conjunction with a "flying focus"—a chromatic focusing system combined with a chirped pump beam that provides spatiotemporal control over the pump's focal spot. Pump intensity isosurfaces are made to propagate at v =-c so as to be in sync with the injected counterpropagating seed pulse. By setting the pump intensity in the interaction region to be just above the ionization threshold of the background gas, an ionization wave is produced that travels at a fixed distance ahead of the seed. Simulations show that this will make it possible to optimize the plasma temperature and mitigate many of the issues that are known to have impacted previous Raman amplification experiments, in particular, the growth of precursors.

  15. Protein Misfolding Cyclic Amplification of Infectious Prions.

    Science.gov (United States)

    Moda, Fabio

    2017-01-01

    Transmissible spongiform encephalopathies, or prion diseases, are a group of incurable disorders caused by the accumulation of an abnormally folded prion protein (PrP Sc ) in the brain. According to the "protein-only" hypothesis, PrP Sc is the infectious agent able to propagate the disease by acting as a template for the conversion of the correctly folded prion protein (PrP C ) into the pathological isoform. Recently, the mechanism of PrP C conversion has been mimicked in vitro using an innovative technique named protein misfolding cyclic amplification (PMCA). This technology represents a great tool for studying diverse aspects of prion biology in the field of basic research and diagnosis. Moreover, PMCA can be expanded for the study of the misfolding process associated to other neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, and frontotemporal lobar degeneration. © 2017 Elsevier Inc. All rights reserved.

  16. Markovian Dynamics of Josephson Parametric Amplification

    Directory of Open Access Journals (Sweden)

    W. Kaiser

    2017-09-01

    Full Text Available In this work, we derive the dynamics of the lossy DC pumped non-degenerate Josephson parametric amplifier (DCPJPA. The main element in a DCPJPA is the superconducting Josephson junction. The DC bias generates the AC Josephson current varying the nonlinear inductance of the junction. By this way the Josephson junction acts as the pump oscillator as well as the time varying reactance of the parametric amplifier. In quantum-limited amplification, losses and noise have an increased impact on the characteristics of an amplifier. We outline the classical model of the lossy DCPJPA and derive the available noise power spectral densities. A classical treatment is not capable of including properties like spontaneous emission which is mandatory in case of amplification at the quantum limit. Thus, we derive a quantum mechanical model of the lossy DCPJPA. Thermal losses are modeled by the quantum Langevin approach, by coupling the quantized system to a photon heat bath in thermodynamic equilibrium. The mode occupation in the bath follows the Bose-Einstein statistics. Based on the second quantization formalism, we derive the Heisenberg equations of motion of both resonator modes. We assume the dynamics of the system to follow the Markovian approximation, i.e. the system only depends on its actual state and is memory-free. We explicitly compute the time evolution of the contributions to the signal mode energy and give numeric examples based on different damping and coupling constants. Our analytic results show, that this model is capable of including thermal noise into the description of the DC pumped non-degenerate Josephson parametric amplifier.

  17. Markovian Dynamics of Josephson Parametric Amplification

    Science.gov (United States)

    Kaiser, Waldemar; Haider, Michael; Russer, Johannes A.; Russer, Peter; Jirauschek, Christian

    2017-09-01

    In this work, we derive the dynamics of the lossy DC pumped non-degenerate Josephson parametric amplifier (DCPJPA). The main element in a DCPJPA is the superconducting Josephson junction. The DC bias generates the AC Josephson current varying the nonlinear inductance of the junction. By this way the Josephson junction acts as the pump oscillator as well as the time varying reactance of the parametric amplifier. In quantum-limited amplification, losses and noise have an increased impact on the characteristics of an amplifier. We outline the classical model of the lossy DCPJPA and derive the available noise power spectral densities. A classical treatment is not capable of including properties like spontaneous emission which is mandatory in case of amplification at the quantum limit. Thus, we derive a quantum mechanical model of the lossy DCPJPA. Thermal losses are modeled by the quantum Langevin approach, by coupling the quantized system to a photon heat bath in thermodynamic equilibrium. The mode occupation in the bath follows the Bose-Einstein statistics. Based on the second quantization formalism, we derive the Heisenberg equations of motion of both resonator modes. We assume the dynamics of the system to follow the Markovian approximation, i.e. the system only depends on its actual state and is memory-free. We explicitly compute the time evolution of the contributions to the signal mode energy and give numeric examples based on different damping and coupling constants. Our analytic results show, that this model is capable of including thermal noise into the description of the DC pumped non-degenerate Josephson parametric amplifier.

  18. Parametric amplifications in the nonlinear transmission line

    Energy Technology Data Exchange (ETDEWEB)

    Kawata, T; Sakai, J; Inoue, H [Toyama Univ., Takaoka (Japan). Faculty of Engineering

    1980-03-01

    The parametric amplification in a transmission line with nonlinear capacitors is analysed theoretically using the equations of three wave interactions. Since this line has two modes, high frequency and low frequency modes, there may occur some mode coupling phenomena through the resonant interactions. We consider three waves with wave number k sub(j) and frequency ..omega..sub(j) in resonance with each other, that is, ..omega../sub 1/ + ..omega../sub 2/ = ..omega../sub 3/ and k/sub 1/ + k/sub 2/ = k/sub 3/, where 0 <= ..omega../sub 1/ <= ..omega../sub 2/ <= ..omega../sub 3/ and k/sub 3/ >= 0. Such conditions are realized in our network and there exist two states: ''forward state'' (each group velocity is positive) and ''backward state'' (one of the group velocities is negative). The coupled equations of three waves has two constant pumps: high frequency (HF) pump and low frequency (LF) pump. Using linear approximations, we examine the possible types of parametric amplification and obtain the power gains depending on the frequency deviation. For only the case of HF pump we get the gain between signals with seme frequency and also get the gain from the low frequency signal to the high frequency signal (''up-conversion'') for the LF pump. The nonlinear analysis gives the exact relation between input and output signals. For the forward state the gain is absolutely suppressed by the ratio of pumping power to input power, while the gain of backward state has no finite maximum and there may appear an ''oscillating state'' if the pumping power is comparatively small.

  19. Ground amplification determined from borehole accelerograms

    International Nuclear Information System (INIS)

    Archuleta, R.J.; Seale, S.H.

    1991-01-01

    The Garner Valley downhole array (GVDA) consists of one surface accelerometer and four downhole accelerometers at depths of 6 m, 15 m, 22m, and 220 m. The five, three-component vertical array of dual-gain accelerometers are capable of measuring accelerations from 3 x 10 -6 g to 2.0 g over a frequency range from 0.0 Hz (0.025, high-gain) Hz to 100 Hz. The site (33 degree 41.60' N, 116 degree 40.20 degree W) is only seven kilometers off the trace of the San Jacinto fault, the most active strand of the San Andreas fault system in southern California and only about 35 km from the San Andreas fault itself. Analysis of individual spectra and spectral ratios for the various depths shows that the zone of weathered granite has a pronounced effect on the spectral amplitudes for frequencies greater than 40 Hz. The soil layer impedance may amplify the high frequencies more than it attenuates. This result must be checked more thoroughly with special consideration of the spectra of the P-wave coda on the horizontal components. Analysis of the P-wave spectra and the spectral ratios shows an increased amplification in the same frequency range (60-90 Hz) where the S-wave spectral ratios imply a change in the attenuation. Comparison of acceleration spectra from two earthquakes, M L 4.2 and M L 2.5 that have nearly the same hypocenter, shows that the near surface amplification and attenuation is nearly the same for both earthquakes. However, the earthquakes themselves are different if we can assume that the recording at 220 m reflects the source spectra with a slight attenuation. The M L 2.5 earthquake has significantly greater high frequency content if the spectra are normalized at the low frequency, i.e., normalization by seismic moment

  20. Microfabricated sleeve devices for chemical reactions

    Science.gov (United States)

    Northrup, M. Allen

    2003-01-01

    A silicon-based sleeve type chemical reaction chamber that combines heaters, such as doped polysilicon for heating, and bulk silicon for convection cooling. The reaction chamber combines a critical ratio of silicon and non-silicon based materials to provide the thermal properties desired. For example, the chamber may combine a critical ratio of silicon and silicon nitride to the volume of material to be heated (e.g., a liquid) in order to provide uniform heating, yet low power requirements. The reaction chamber will also allow the introduction of a secondary tube (e.g., plastic) into the reaction sleeve that contains the reaction mixture thereby alleviating any potential materials incompatibility issues. The reaction chamber may be utilized in any chemical reaction system for synthesis or processing of organic, inorganic, or biochemical reactions, such as the polymerase chain reaction (PCR) and/or other DNA reactions, such as the ligase chain reaction, which are examples of a synthetic, thermal-cycling-based reaction. The reaction chamber may also be used in synthesis instruments, particularly those for DNA amplification and synthesis.

  1. Study on high gain broadband optical parametric chirped pulse amplification

    International Nuclear Information System (INIS)

    Zhang, S.K.; Fujita, M.; Yamanaka, C.; Yoshida, H.; Kodama, R.; Fujita, H.; Nakatsuka, M.; Izawa, Y.

    2000-01-01

    Optical parametric chirped pulse amplification has apparent advantages over the current schemes for high energy ultrashort pulse amplification. High gain in a single pass amplification, small B-integral, low heat deposition, high contrast ratio and, especially the extremely broad gain bandwidth with large-size crystals available bring people new hope for over multi-PW level at which the existing Nd:glass systems suffered difficulties. In this paper we present simulation and experimental studies for a high gain optical parametric chirped pulse amplification system which may be used as a preamplifier to replace the current complicated regenerative system or multi-pass Ti:sapphire amplifiers. Investigations on the amplification bandwidth and gain with BBO are performed. Analysis and discussions are also given. (author)

  2. Establishment of recombinase polymerase amplification assay for five hemorrhagic fever-related viruses

    Directory of Open Access Journals (Sweden)

    Xue-feng CAO

    2017-08-01

    Full Text Available Objective To establish a one-step recombinase polymerase amplification (RPA method for pathogen screening and rapid detection in the field targeting for five hemorrhagic fever related viruses (Zaire ebola virus, Sudan ebola virus, Marburg virus, Lassa virus and Yellow fever virus. Methods The specific nucleic acid (NA fragments of each virus were selected as target genes by genome sequence analysis, and the primers and probes for RPA assays were designed according to the sequence. A series of diluted template genes were used for RPA detection to determine the sensitivity. The hemorrhagic fever-related viral nucleic acids were used for RPA detection to determine the specificity. The amplification experiments were carried out at different temperature ranging from 37℃ to 42℃ to validate the reaction temperature range. Results The RPA reaction systems of the five hemorrhagic fever viruses could effectively amplify the target genes, the sensitivities were between 1.5×102 and 1.5×103 copies. No cross reactions existed with the other hemorrhagic fever-related viral genes. Meanwhile, RPA assay could effectively amplify the target genes at 37-42℃. Conclusion The isothermal RPA assays of five hemorrhagic fever viruses are established, which may amply target genes fast and react at a wide temperature range, and be potentially useful for in field pathogens detection. DOI: 10.11855/j.issn.0577-7402.2017.06.09

  3. An Engineered Kinetic Amplification Mechanism for Single Nucleotide Variant Discrimination by DNA Hybridization Probes.

    Science.gov (United States)

    Chen, Sherry Xi; Seelig, Georg

    2016-04-20

    Even a single-nucleotide difference between the sequences of two otherwise identical biological nucleic acids can have dramatic functional consequences. Here, we use model-guided reaction pathway engineering to quantitatively improve the performance of selective hybridization probes in recognizing single nucleotide variants (SNVs). Specifically, we build a detection system that combines discrimination by competition with DNA strand displacement-based catalytic amplification. We show, both mathematically and experimentally, that the single nucleotide selectivity of such a system in binding to single-stranded DNA and RNA is quadratically better than discrimination due to competitive hybridization alone. As an additional benefit the integrated circuit inherits the property of amplification and provides at least 10-fold better sensitivity than standard hybridization probes. Moreover, we demonstrate how the detection mechanism can be tuned such that the detection reaction is agnostic to the position of the SNV within the target sequence. in contrast, prior strand displacement-based probes designed for kinetic discrimination are highly sensitive to position effects. We apply our system to reliably discriminate between different members of the let-7 microRNA family that differ in only a single base position. Our results demonstrate the power of systematic reaction network design to quantitatively improve biotechnology.

  4. Development of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of BK Virus▿

    Science.gov (United States)

    Bista, Bipin Raj; Ishwad, Chandra; Wadowsky, Robert M.; Manna, Pradip; Randhawa, Parmjeet Singh; Gupta, Gaurav; Adhikari, Meena; Tyagi, Rakhi; Gasper, Gina; Vats, Abhay

    2007-01-01

    Loop-mediated isothermal amplification (LAMP) is a novel method for rapid amplification of DNA. Its advantages include rapidity and minimal equipment requirement. The LAMP assay was developed for BK virus (BKV), which is a leading cause of morbidity in renal transplant recipients. The characteristics of the assay, including its specificity and sensitivity, were evaluated. BKV LAMP was performed using various incubation times with a variety of specimens, including unprocessed urine and plasma samples. A ladder pattern on gel electrophoresis, typical of successful LAMP reactions, was observed specifically only for BKV and not for other viruses. The sensitivity of the assay with 1 h of incubation was 100 copies/tube of a cloned BKV fragment. Additionally, a positive reaction was visually ascertained by a simple color reaction using SYBR green dye. BKV LAMP was also successful for urine and plasma specimens without the need for DNA extraction. Due to its simplicity and specificity, the LAMP assay can potentially be developed for “point of care” screening of BKV. PMID:17314224

  5. Development of a loop-mediated isothermal amplification assay for rapid detection of BK virus.

    Science.gov (United States)

    Bista, Bipin Raj; Ishwad, Chandra; Wadowsky, Robert M; Manna, Pradip; Randhawa, Parmjeet Singh; Gupta, Gaurav; Adhikari, Meena; Tyagi, Rakhi; Gasper, Gina; Vats, Abhay

    2007-05-01

    Loop-mediated isothermal amplification (LAMP) is a novel method for rapid amplification of DNA. Its advantages include rapidity and minimal equipment requirement. The LAMP assay was developed for BK virus (BKV), which is a leading cause of morbidity in renal transplant recipients. The characteristics of the assay, including its specificity and sensitivity, were evaluated. BKV LAMP was performed using various incubation times with a variety of specimens, including unprocessed urine and plasma samples. A ladder pattern on gel electrophoresis, typical of successful LAMP reactions, was observed specifically only for BKV and not for other viruses. The sensitivity of the assay with 1 h of incubation was 100 copies/tube of a cloned BKV fragment. Additionally, a positive reaction was visually ascertained by a simple color reaction using SYBR green dye. BKV LAMP was also successful for urine and plasma specimens without the need for DNA extraction. Due to its simplicity and specificity, the LAMP assay can potentially be developed for "point of care" screening of BKV.

  6. Influence of Calcium Chloride (CaCl/sub 2/) on Fruit Quality of Pear (Pyrus communis) cv. le conte During Storage

    International Nuclear Information System (INIS)

    Sajid, M.; Mukhtiar, M.; Rab, M.; Shah, S.R.; Jan, I.

    2014-01-01

    An experiment was conducted at Postharvest laboratory, Department of Horticulture, The University of Agriculture Peshawar, during 2010 to evaluate the 'Influence of calcium chloride (CaCl/sub 2/) on fruit quality of pear (Pyrus communis) cv. Le-conte'. The experiment was laid out in Randomized Complete Block Design (RCBD) with three factors i.e. (CaCl/sub 2/) concentration (0,3,6 and 9%), dipping time (5,10 and 15 minutes) and storage durations (0,10,20,30 and 40 days). The maximum ascorbic acid (7.93 mg/100 g), reducing sugar (5.86%), while the least percent weight loss (4.52%), pH of fruit juice (4.42), total soluble solids (TSS) (19.83%) and percent disease incidence (2.56%) were observed in fruits treated with 9% CaCl/sub 2/ solution. The dipping time also significantly influenced the quality attributes of pear fruits during storage. The more ascorbic acid (6.88 mg/100 g) and reducing sugar (5.44%) were recorded in the pear fruits dipped for 15 minutes. Storage duration significantly affected the fruit quality 40 days storage. The highest reducing sugar recorded in fresh pear fruits while the highest ascorbic acid (7.83 mg/100 g) were observed in pear fruit stored for 10 days storage duration, while more non-reducing sugar (7.03%) recorded in the fruits stored for 30 days. In the interaction of CaCl/sub 2/ concentration x dipping time, the highest total soluble solid (31.88%) noted in the fruits stored for 40 days and dipped for 5 minutes in CaCl/sub 2/ solution. It is concluded that pear fruit perform best in the postharvest life when treated with 9% CaCl/sub 2/ solution and dipped for 15 minutes. It retained most of the quality attributes up to 10 days storage at ambient temperature while a significant decline was recorded in fruit quality when extended the storage duration from 20 to 40 days. (author)

  7. A Novel Low Temperature PCR Assured High-Fidelity DNA Amplification

    Directory of Open Access Journals (Sweden)

    Shaoxia Zhou

    2013-06-01

    Full Text Available As previously reported, a novel low temperature (LoTemp polymerase chain reaction (PCR catalyzed by a moderately heat-resistant (MHR DNA polymerase with a chemical-assisted denaturation temperature set at 85 °C instead of the conventional 94–96 °C can achieve high-fidelity DNA amplification of a target DNA, even after up to 120 PCR thermal cycles. Furthermore, such accurate amplification is not achievable with conventional PCR. Now, using a well-recognized L1 gene segment of the human papillomavirus (HPV type 52 (HPV-52 as the template for experiments, we demonstrate that the LoTemp high-fidelity DNA amplification is attributed to an unusually high processivity and stability of the MHR DNA polymerase whose high fidelity in template-directed DNA synthesis is independent of non-existent 3'–5' exonuclease activity. Further studies and understanding of the characteristics of the LoTemp PCR technology may facilitate implementation of DNA sequencing-based diagnostics at the point of care in community hospital laboratories.

  8. A quantitative comparison of single-cell whole genome amplification methods.

    Directory of Open Access Journals (Sweden)

    Charles F A de Bourcy

    Full Text Available Single-cell sequencing is emerging as an important tool for studies of genomic heterogeneity. Whole genome amplification (WGA is a key step in single-cell sequencing workflows and a multitude of methods have been introduced. Here, we compare three state-of-the-art methods on both bulk and single-cell samples of E. coli DNA: Multiple Displacement Amplification (MDA, Multiple Annealing and Looping Based Amplification Cycles (MALBAC, and the PicoPLEX single-cell WGA kit (NEB-WGA. We considered the effects of reaction gain on coverage uniformity, error rates and the level of background contamination. We compared the suitability of the different WGA methods for the detection of copy-number variations, for the detection of single-nucleotide polymorphisms and for de-novo genome assembly. No single method performed best across all criteria and significant differences in characteristics were observed; the choice of which amplifier to use will depend strongly on the details of the type of question being asked in any given experiment.

  9. Massively parallel whole genome amplification for single-cell sequencing using droplet microfluidics.

    Science.gov (United States)

    Hosokawa, Masahito; Nishikawa, Yohei; Kogawa, Masato; Takeyama, Haruko

    2017-07-12

    Massively parallel single-cell genome sequencing is required to further understand genetic diversities in complex biological systems. Whole genome amplification (WGA) is the first step for single-cell sequencing, but its throughput and accuracy are insufficient in conventional reaction platforms. Here, we introduce single droplet multiple displacement amplification (sd-MDA), a method that enables massively parallel amplification of single cell genomes while maintaining sequence accuracy and specificity. Tens of thousands of single cells are compartmentalized in millions of picoliter droplets and then subjected to lysis and WGA by passive droplet fusion in microfluidic channels. Because single cells are isolated in compartments, their genomes are amplified to saturation without contamination. This enables the high-throughput acquisition of contamination-free and cell specific sequence reads from single cells (21,000 single-cells/h), resulting in enhancement of the sequence data quality compared to conventional methods. This method allowed WGA of both single bacterial cells and human cancer cells. The obtained sequencing coverage rivals those of conventional techniques with superior sequence quality. In addition, we also demonstrate de novo assembly of uncultured soil bacteria and obtain draft genomes from single cell sequencing. This sd-MDA is promising for flexible and scalable use in single-cell sequencing.

  10. Detection of genetically modified organisms (GMOs using isothermal amplification of target DNA sequences

    Directory of Open Access Journals (Sweden)

    La Mura Maurizio

    2009-02-01

    Full Text Available Abstract Background The most common method of GMO detection is based upon the amplification of GMO-specific DNA amplicons using the polymerase chain reaction (PCR. Here we have applied the loop-mediated isothermal amplification (LAMP method to amplify GMO-related DNA sequences, 'internal' commonly-used motifs for controlling transgene expression and event-specific (plant-transgene junctions. Results We have tested the specificity and sensitivity of the technique for use in GMO studies. Results show that detection of 0.01% GMO in equivalent background DNA was possible and dilutions of template suggest that detection from single copies of the template may be possible using LAMP. Conclusion This work shows that GMO detection can be carried out using LAMP for routine screening as well as for specific events detection. Moreover, the sensitivity and ability to amplify targets, even with a high background of DNA, here demonstrated, highlights the advantages of this isothermal amplification when applied for GMO detection.

  11. Duplex recombinase polymerase amplification assays incorporating competitive internal controls for bacterial meningitis detection.

    Science.gov (United States)

    Higgins, Owen; Clancy, Eoin; Forrest, Matthew S; Piepenburg, Olaf; Cormican, Martin; Boo, Teck Wee; O'Sullivan, Nicola; McGuinness, Claire; Cafferty, Deirdre; Cunney, Robert; Smith, Terry J

    2018-04-01

    Recombinase polymerase amplification (RPA) is an isothermal nucleic acid amplification technology that provides rapid and robust infectious disease pathogen detection, ideal for point-of-care (POC) diagnostics in disease-prevalent low-resource countries. We have developed and evaluated three duplex RPA assays incorporating competitive internal controls for the detection of leading bacterial meningitis pathogens. Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae singleplex RPA assays were initially developed and evaluated, demonstrating 100% specificity with limits of detection of 4.1, 8.5 and 3.9 genome copies per reaction, respectively. Each assay was further developed into internally controlled duplex RPA assays via the incorporation of internal amplification control templates. Clinical performance of each internally controlled duplex RPA assay was evaluated by testing 64 archived PCR-positive clinical samples. Compared to real-time PCR, all duplex RPA assays demonstrated 100% diagnostic specificity, with diagnostic sensitivities of 100%, 86.3% and 100% for the S. pneumoniae, N. meningitidis and H. influenzae assays, respectively. This study details the first report of internally controlled duplex RPA assays for the detection of bacterial meningitis pathogens: S. pneumoniae, N. meningitidis and H. influenzae. We have successfully demonstrated the clinical diagnostic utility of each duplex RPA assay, introducing effective diagnostic technology for POC bacterial meningitis identification in disease-prevalent developing countries. Copyright © 2018 Elsevier Inc. All rights reserved.

  12. Amplification volume reduction on DNA database samples using FTA™ Classic Cards.

    Science.gov (United States)

    Wong, Hang Yee; Lim, Eng Seng Simon; Tan-Siew, Wai Fun

    2012-03-01

    The DNA forensic community always strives towards improvements in aspects such as sensitivity, robustness, and efficacy balanced with cost efficiency. Therefore our laboratory decided to study the feasibility of PCR amplification volume reduction using DNA entrapped in FTA™ Classic Card and to bring cost savings to the laboratory. There were a few concerns the laboratory needed to address. First, the kinetics of the amplification reaction could be significantly altered. Second, an increase in sensitivity might affect interpretation due to increased stochastic effects even though they were pristine samples. Third, statics might cause FTA punches to jump out of its allocated well into another thus causing sample-to-sample contamination. Fourth, the size of the punches might be too small for visual inspection. Last, there would be a limit to the extent of volume reduction due to evaporation and the possible need of re-injection of samples for capillary electrophoresis. The laboratory had successfully optimized a reduced amplification volume of 10 μL for FTA samples. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  13. Clinical application of somatosensory amplification in psychosomatic medicine

    Directory of Open Access Journals (Sweden)

    Nakao Mutsuhiro

    2007-10-01

    Full Text Available Abstract Many patients with somatoform disorders are frequently encountered in psychosomatic clinics as well as in primary care clinics. To assess such patients objectively, the concept of somatosensory amplification may be useful. Somatosensory amplification refers to the tendency to experience a somatic sensation as intense, noxious, and disturbing. It may have a role in a variety of medical conditions characterized by somatic symptoms that are disproportionate to demonstrable organ pathology. It may also explain some of the variability in somatic symptomatology found among different patients with the same serious medical disorder. It has been assessed with a self-report questionnaire, the Somatosensory Amplification Scale. This instrument was developed in a clinical setting in the U.S., and the reliability and validity of the Japanese and Turkish versions have been confirmed as well. Many studies have attempted to clarify the specific role of somatosensory amplification as a pathogenic mechanism in somatization. It has been reported that somatosensory amplification does not correlate with heightened sensitivity to bodily sensations and that emotional reactivity exerts its influence on somatization via a negatively biased reporting style. According to our recent electroencephalographic study, somatosensory amplification appears to reflect some aspects of long-latency cognitive processing rather than short-latency interoceptive sensitivity. The concept of somatosensory amplification can be useful as an indicator of somatization in the therapy of a broad range of disorders, from impaired self-awareness to various psychiatric disorders. It also provides useful information for choosing appropriate pharmacological or psychological therapy. While somatosensory amplification has a role in the presentation of somatic symptoms, it is closely associated with other factors, namely, anxiety, depression, and alexithymia that may also influence the same

  14. Nuclear reactions

    International Nuclear Information System (INIS)

    Lane, A.M.

    1980-01-01

    In reviewing work at Harwell over the past 25 years on nuclear reactions it is stated that a balance has to be struck in both experiment and theory between work on cross-sections of direct practical relevance to reactors and on those relevant to an overall understanding of reaction processes. The compound nucleus and direct process reactions are described. Having listed the contributions from AERE, Harwell to developments in nuclear reaction research in the period, work on the optical model, neutron capture theory, reactions at doorway states with fine structure, and sum-rules for spectroscopic factors are considered in more detail. (UK)

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

    Directory of Open Access Journals (Sweden)

    Siwon Lee

    2015-12-01

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

  16. Development of Loop-Mediated Isothermal Amplification Assay for Detection of Entamoeba histolytica▿

    Science.gov (United States)

    Liang, Shih-Yu; Chan, Yun-Hsien; Hsia, Kan-Tai; Lee, Jing-Lun; Kuo, Ming-Chu; Hwa, Kuo-Yuan; Chan, Chi-Wen; Chiang, Ting-Yi; Chen, Jung-Sheng; Wu, Fang-Tzy; Ji, Dar-Der

    2009-01-01

    A novel one-step, closed-tube, loop-mediated isothermal amplification (LAMP) assay for detecting Entamoeba histolytica, one of the leading causes of morbidity in developing countries, was developed. The sensitivity of the LAMP assay is 1 parasite per reaction. A total of 130 clinical samples were analyzed, and the results compared with those of conventional nested PCR to validate the practicability of this assay. No DNA was amplified from other diarrheal pathogens, such as other Entamoeba species, bacteria, and viruses. These results indicate that LAMP is a rapid, simple, and valuable diagnostic tool for epidemiological studies of amebiasis. PMID:19321720

  17. Rapid PCR amplification using a microfluidic device with integrated microwave heating and air impingement cooling.

    Science.gov (United States)

    Shaw, Kirsty J; Docker, Peter T; Yelland, John V; Dyer, Charlotte E; Greenman, John; Greenway, Gillian M; Haswell, Stephen J

    2010-07-07

    A microwave heating system is described for performing polymerase chain reaction (PCR) in a microfluidic device. The heating system, in combination with air impingement cooling, provided rapid thermal cycling with heating and cooling rates of up to 65 degrees C s(-1) and minimal over- or under-shoot (+/-0.1 degrees C) when reaching target temperatures. In addition, once the required temperature was reached it could be maintained with an accuracy of +/-0.1 degrees C. To demonstrate the functionality of the system, PCR was successfully performed for the amplification of the Amelogenin locus using heating rates and quantities an order of magnitude faster and smaller than current commercial instruments.

  18. Rapid and sensitive detection of canine parvovirus type 2 by recombinase polymerase amplification.

    Science.gov (United States)

    Wang, Jianchang; Liu, Libing; Li, Ruiwen; Wang, Jinfeng; Fu, Qi; Yuan, Wanzhe

    2016-04-01

    A novel recombinase polymerase amplification (RPA)-based method for detection of canine parvovirus type 2 (CPV-2) was developed. Sensitivity analysis showed that the detection limit of RPA was 10 copies of CPV-2 genomic DNA. RPA amplified both CPV-2a and -2b DNA but did not amplify the template of other important dog viruses (CCoV, PRV or CDV), demonstrating high specificity. The method was further validated with 57 canine fecal samples. An outstanding advantage of RPA is that it is an isothermal reaction and can be performed in a water bath, making RPA a potential alternative method for CPV-2 detection in resource-limited settings.

  19. Loop-mediated isothermal amplification (LAMP) shield for Arduino DNA detection.

    Science.gov (United States)

    Velders, Aldrik H; Schoen, Cor; Saggiomo, Vittorio

    2018-02-01

    Loop-mediated isothermal amplification (LAMP) of DNA is gaining relevance as a method to detect nucleic acids, as it is easier, faster, and more powerful than conventional Polymerase Chain Reaction. However, LAMP is still mostly used in laboratory settings, because of the lack of a cheap and easy, one-button device that can perform LAMP experiments. Here we show how to build and program an Arduino shield for a LAMP and detection of DNA. The here described Arduino Shield is cheap, easy to assemble, to program and use, it is battery operated and the detection of DNA is done by naked-eye so that it can be used in field.

  20. Development, validation, and standardization of polymerase chain reaction-based detection of E-coli O157

    DEFF Research Database (Denmark)

    Abdulmawjood, A.; Bulte, M.; Roth, S.

    2004-01-01

    A diagnostic polymerase chain reaction assay was developed for the detection of E. coli O157 as the first part of a multicenter validation and standardization project. The assay is based on amplification of sequences of the rfbE O157 gene and includes an internal amplification control. The select...