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Sample records for amplification reaction pear

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

    Directory of Open Access Journals (Sweden)

    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. Nucleic acid amplification: Alternative methods of polymerase chain reaction.

    Science.gov (United States)

    Fakruddin, Md; Mannan, Khanjada Shahnewaj Bin; Chowdhury, Abhijit; Mazumdar, Reaz Mohammad; Hossain, Md Nur; Islam, Sumaiya; Chowdhury, Md Alimuddin

    2013-10-01

    Nucleic acid amplification is a valuable molecular tool not only in basic research but also in application oriented fields, such as clinical medicine development, infectious diseases diagnosis, gene cloning and industrial quality control. A comperehensive review of the literature on the principles, applications, challenges and prospects of different alternative methods of polymerase chain reaction (PCR) was performed. PCR was the first nucleic acid amplification method. With the advancement of research, a no of alternative nucleic acid amplification methods has been developed such as loop mediated isothermal amplification, nucleic acid sequence based amplification, strand displacement amplification, multiple displacement amplification. Most of the alternative methods are isothermal obviating the need for thermal cyclers. Though principles of most of the alternate methods are relatively complex than that of PCR, they offer better applicability and sensitivity in cases where PCR has limitations. Most of the alternate methods still have to prove themselves through extensive validation studies and are not available in commercial form; they pose the potentiality to be used as replacements of PCR. Continuous research is going on in different parts of the world to make these methods viable technically and economically.

  3. Duplex real-time polymerase chain reaction reveals competition between Erwinia amylovora and E. pyrifoliae on pear blossoms.

    Science.gov (United States)

    Lehman, Susan M; Kim, Won-Sik; Castle, Alan J; Svircev, Antonet M

    2008-06-01

    Erwinia amylovora and E. pyrifoliae are the causative agents of fire blight and Asian pear blight, respectively. The pathogens are closely related, with overlapping host ranges. Data are unavailable on the current distribution of E. pyrifoliae and on the interaction between the two species when they are present together on the same host. In this study, a duplex real-time polymerase chain reaction (PCR) protocol was developed to monitor the population dynamics of E. amylovora and E. pyrifoliae on the surface of Bartlett pear blossoms. Bacterial cells washed from blossoms were used directly as the PCR template without DNA extraction. Primers and a probe based on the E. amylovora levansucrase gene detected all E. amylovora strains. All E. pyrifoliae strains, including the Japanese Erwinia strains previously described as E. amylovora, were detected with a primer and probe combination based on the E. pyrifoliae hrpW gene. Disease development and severity were not significantly different in blossoms inoculated with individual Erwinia species or with a mixture of the two species. However, E. amylovora grew to greater population sizes than did E. pyrifoliae in both single species inoculations and in mixtures, suggesting that E. amylovora has a greater competitive fitness on Bartlett pear blossoms than E. pyrifoliae.

  4. Polymerase chain reaction amplification of genomic fragments of bovine herpesvirus-1

    OpenAIRE

    Cândido AL; ED Bontempo; Resende M.

    2000-01-01

    Especial conditions were developed for the amplification of five DNA segments from US region of BHV-1 by polymerase chain reaction. In order to eliminate most nonspecific products it was found that addition of three cosolvents DMSO, glycerol and NP 40 was a simple method for increasing the specificity of amplification.

  5. Thermal isolation of microchip reaction chambers for rapid non-contact DNA amplification

    Science.gov (United States)

    Easley, Christopher J.; Humphrey, Joseph A. C.; Landers, James P.

    2007-09-01

    This paper describes further optimization of a non-contact, infrared-mediated system for microchip DNA amplification via the polymerase chain reaction (PCR). The optimization is focused on heat transfer modeling and subsequent fabrication of thermally isolated reaction chambers in glass devices that are uniquely compatible with non-contact thermal control. With a thermal conductivity approximately an order of magnitude higher than many plastics, glass is not the obvious substrate of choice for rapid thermal cycling in microfluidic chambers, yet it is preferable in terms of optical clarity, solvent compatibility and chemical inertness. Based on predictions of a lumped capacity heat transfer analysis, it is shown here that post-bonding, patterned etching of surrounding glass from microfluidic reaction chambers provides enhancements as high as 3.6- and 7.5-fold in cooling and heating rates, respectively, over control devices with the same chamber designs. These devices are then proven functional for rapid DNA amplification via PCR, in which 25 thermal cycles are completed in only 5 min in thermally isolated PCR chambers of 270 nL volume, representing the fastest static PCR in glass devices reported to date. Amplification of the 500-base pair fragment of λ-DNA was confirmed by capillary gel electrophoresis. In addition to rapid temperature control, the fabrication scheme presented, which is compatible with standard photolithography and wet etching techniques, provides a simple alternative for general thermal management in glass microfluidic devices without metallization.

  6. 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.

  7. 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.

  8. Study of differential polymerase chain reaction of C-erbB-2 oncogene amplification in gastric cancer

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    AIM To study the significance of C-erbB-2 oncogene amplification in gastric cancer.METHODS C-erbB-2 oncogene amplification was examined by using differential polymerase chain reaction (dPCR) in surgical and endoscopic specimens of 83 cases of gastric cancer and 101 metastatic lymph nodes.RESULTS C-erbB-2 amplification was found in 28.9% (24/ 83) surgical specimens and 20.5% (17/ 83) endoscopic ones of gastric cancer patients. The amplification was significant in both types of specimens of advanced cancer cases (P<0.05) and surgical specimens with lymph node metastasis (P<0.01). The incidence of C-erbB-2 amplification in lymph nodes with metastasis was higher than in primary sites (surgical specimens, P<0.05). The patients with amplification tumors had poorer 5-year survival rates than those with unamplification ones in the early cancers and well to moderately differentiated adenocarcinomas (P<0.05). The same surgical samples were tested again by Southern blot hybridization to ascertain C-erbB-2 amplification, and the positive rate of C-erbB-2 amplification (15.7%) was lower than that of dPCR (28.9%, P<0.05).CONCLUSION Examining C-erbB-2 amplification by dPCR is a quick, simple, reliable and independent method, and is helpful in predicting prognosis and metastatic potential of gastric cancer.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. Nuclemeter: A Reaction-Diffusion Column for Quantifying Nucleic Acids Undergoing Enzymatic Amplification

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    Bau, Haim; Liu, Changchun; Killawala, Chitvan; Sadik, Mohamed; Mauk, Michael

    2014-11-01

    Real-time amplification and quantification of specific nucleic acid sequences plays a major role in many medical and biotechnological applications. In the case of infectious diseases, quantification of the pathogen-load in patient specimens is critical to assessing disease progression, effectiveness of drug therapy, and emergence of drug-resistance. Typically, nucleic acid quantification requires sophisticated and expensive instruments, such as real-time PCR machines, which are not appropriate for on-site use and for low resource settings. We describe a simple, low-cost, reactiondiffusion based method for end-point quantification of target nucleic acids undergoing enzymatic amplification. The number of target molecules is inferred from the position of the reaction-diffusion front, analogous to reading temperature in a mercury thermometer. We model the process with the Fisher Kolmogoroff Petrovskii Piscounoff (FKPP) Equation and compare theoretical predictions with experimental observations. The proposed method is suitable for nucleic acid quantification at the point of care, compatible with multiplexing and high-throughput processing, and can function instrument-free. C.L. was supported by NIH/NIAID K25AI099160; M.S. was supported by the Pennsylvania Ben Franklin Technology Development Authority; C.K. and H.B. were funded, in part, by NIH/NIAID 1R41AI104418-01A1.

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

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    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.

  12. Single primer amplification reaction methods reveal exotic and indigenous mulberry varieties are similarly diverse

    Indian Academy of Sciences (India)

    Esha Bhattacharya; S B Dandin; Shirish Anand Ranade

    2005-12-01

    Mulberry is the sole food source for mulberry silkworm and a number of indigenous and exotic varieties are used in sericulture. Studies on assessment of genetic diversity have been done amongst a few mulberry varieties using one or at the most two methods. However, no comprehensive study on a large number of varieties has been carried out. In present study, single primer amplification reaction (SPAR) methods have been used for determination of diversity in 27 mulberry varieties (exotic as well as indigenous), using four minisatellite core sequence primers for directed amplification of minisatellite DNA (DAMD), three simple sequence repeat (SSR) motifs as primers for inter simple sequence repeat (ISSR) and 20 arbitrary sequence decamer primers for random amplified polymorphic DNA (RAPD) reactions. The Jaccard coefficients were determined for the DAMD, ISSR and RAPD band data (total of 58, 39 and 235 bands respectively). All three methods revealed wide range of distances supporting a wide range of mulberry genetic diversity. A cumulative analysis of the data generated by three methods resulted in a neighbour-joining (NJ) tree that gave a better reflection of the relatedness and affinities of the varieties to each other. Comparison of the three methods by marker indices and the Mantel test of correlation indicated that though all methods were useful for the assessment of diversity in mulberry, the DAMD method was better. When considered as two groups (10 exotic and 17 indigenous varieties), the mulberry varieties in the exotic group were found to have slightly greater diversity than the indigenous ones. These results support the concept of naturalization of mulberry varieties at locales distant from their origins.

  13. Comparison of DNA extraction methods for polymerase chain reaction amplification of guanaco (Lama guanicoe) fecal DNA samples.

    Science.gov (United States)

    Espinosa, M I; Bertin, A; Squeo, F A; Cortés, A; Gouin, N

    2015-01-23

    Feces-based population genetic studies have become increasingly popular. However, polymerase chain reaction (PCR) amplification rates from fecal material vary depending on the species, populations, loci, and extraction protocols. Here, we assessed the PCR amplification success of three microsatellite markers and a segment of the mitochondrial control region of DNA extracted from field-collected feces of guanaco (Lama guanicoe) using two protocols - Qiagen DNA Stool Kit and 2 cetyltrimethylammonium bromide/phenol:chloroform:isoamyl alcohol (2CTAB/PCI) method. Chelex resin treatment to remove inhibitors was also tested. Our results show that the mitochondrial locus was the most difficult to amplify. PCR success rates improved for all markers after Chelex treatment of extracted DNA, and 2CTAB/PCI method (95.83%) appeared to perform slightly better than stool kit (91.67%) for the nuclear markers. Amplification success was significantly influenced by the extraction method, Chelex treatment, and locus (P 0.89), but they decreased slightly after treatment for amplification of nuclear markers and markedly after treatment for amplification of the mitochondrial control region. Thus, we showed that Chelex treatment gives high PCR success, especially for nuclear markers, and adequate DNA extraction rates can be achieved from L. guanicoe feces even from non-fresh fecal material. Although not significant, 2CTAB/PCI method tended to provide higher successful amplification rates on a whole set of samples, suggesting that the method could be particularly useful when using small sample sizes.

  14. Electrical detection of dsDNA and polymerase chain reaction amplification.

    Science.gov (United States)

    Salm, Eric; Liu, Yi-Shao; Marchwiany, Daniel; Morisette, Dallas; He, Yiping; Razouk, Laila; Bhunia, Arun K; Bashir, Rashid

    2011-12-01

    Food-borne pathogens and food safety-related outbreaks have come to the forefront over recent years. Estimates on the annual cost of sicknesses, hospitalizations, and deaths run into the billions of dollars. There is a large body of research on detection of food-borne pathogens; however, the widely accepted current systems are limited by costly reagents, lengthy time to completion, and expensive equipment. Our aim is to develop a label-free method for determining a change in DNA concentration after a PCR assay. We first used impedance spectroscopy to characterize the change in concentration of purified DNA in deionized water within a microfluidic biochip. To adequately measure the change in DNA concentration in PCR solution, it was necessary to go through a purification and precipitation step to minimize the effects of primers, PCR reagents, and excess salts. It was then shown that the purification and precipitation of the fully amplified PCR reaction showed results similar to the control tests performed with DNA in deionized water. We believe that this work has brought label free electrical biosensors for PCR amplification one step closer to reality.

  15. Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene.

    Science.gov (United States)

    Brakstad, O G; Aasbakk, K; Maeland, J A

    1992-07-01

    Synthetic oligonucleotide primers of 21 and 24 bases, respectively, were used in the polymerase chain reaction (PCR) to amplify a sequence of the nuc gene, which encodes the thermostable nuclease of Staphylococcus aureus. A DNA fragment of approximately 270 bp was amplified from lysed S. aureus cells or isolated DNA. The PCR product was detected by agarose gel electrophoresis or Southern blot analysis by using a 33-mer internal nuc gene hybridization probe. With S. aureus cells the lower detection limit was less than 10 CFU, and with the isolated target the lower detection limit was 0.69 pg of DNA. The primers recognized 90 of 90 reference or clinical S. aureus strains. Amplification was not recorded when 80 strains representing 16 other staphylococcal species were tested or when 20 strains representing 9 different nonstaphylococcal species were tested. Some of the non-S. aureus staphylococci produced thermostable nucleases but were PCR negative. The PCR product was generated when in vitro-cultured S. aureus was used to prepare simulated clinical specimens of blood, urine, cerebrospinal fluid, or synovial fluid. No PCR product was generated when the sterile body fluids were tested. However, the sensitivity of the PCR was reduced when S. aureus in blood or urine was tested in comparison with that when bacteria in saline were tested. With the bacteria in blood, the detection limit of the PCR was 10(3) CFU. A positive PCR result was recorded when a limited number of clinical samples from wounds verified to be infected with S. aureus were tested, while the PCR product was not detected in materials from infections caused by other bacteria. Generation of PCR products was not affected by exposure of S. aureus to bactericidal agents, including cloxacillin and gentamicin, prior to testing, but was affected by exposure to UV radiation. The PCR for amplification of the nuc gene has potential for the rapid diagnosis of S. aureus infections by direct testing of clinical

  16. Reverse transcription genome exponential amplification reaction assay for rapid and universal detection of human rhinoviruses.

    Science.gov (United States)

    Guan, Li; Zhao, Lin-Qing; Zhou, Hang-Yu; Nie, Kai; Li, Xin-Na; Zhang, Dan; Song, Juan; Qian, Yuan; Ma, Xue-Jun

    2016-07-01

    Human rhinoviruses (HRVs) have long been recognized as the cause of more than one-half of acute viral upper respiratory illnesses, and they are associated with more-serious diseases in children, such as asthma, acute otitis media and pneumonia. A rapid and universal test for of HRV infection is in high demand. In this study, a reverse transcription genome exponential amplification reaction (RT-GEAR) assay targeting the HRV 5' untranslated region (UTR) was developed for pan-HRV detection. The reaction was performed in a single tube in one step at 65 °C for 60 min using a real-time fluorometer (Genie(®)II; Optigene). The RT-GEAR assay showed no cross-reactivity with common human enteroviruses, including HEV71, CVA16, CVA6, CVA10, CVA24, CVB5, Echo30, and PV1-3 or with other common respiratory viruses including FluA H3, FluB, PIV1-4, ADV3, RSVA, RSVB and HMPV. With in vitro-transcribed RNA containing the amplified regions of HRV-A60, HRV-B06 and HRV-C07 as templates, the sensitivity of the RT-GEAR assay was 5, 50 and 5 copies/reaction, respectively. Experiments to evaluate the clinical performance of the RT-GEAR assay were also carried out with a panel of 143 previously verified samples, and the results were compared with those obtained using a published semi-nested PCR assay followed by sequencing. The tested panel comprised 91 HRV-negative samples and 52 HRV-positive samples (18 HRV-A-positive samples, 3 HRV-B-positive samples and 31 HRV-C-positive samples). The sensitivity and specificity of the pan-HRVs RT-GEAR assay was 98.08 % and 100 %, respectively. The kappa correlation between the two methods was 0.985. The RT-GEAR assay based on a portable Genie(®)II fluorometer is a sensitive, specific and rapid assay for the universal detection of HRV infection.

  17. Sensitive and specific colorimetric DNA detection by invasive reaction coupled with nicking endonuclease-assisted nanoparticles amplification.

    Science.gov (United States)

    Zou, Bingjie; Cao, Xiaomei; Wu, Haiping; Song, Qinxin; Wang, Jianping; Kajiyama, Tomoharu; Kambara, Hideki; Zhou, Guohua

    2015-04-15

    Colorimetric DNA detection is preferable to methods in clinical molecular diagnostics, because no expensive equipment is required. Although many gold nanoparticle-based colorimetric DNA detection strategies have been developed to analyze DNA sequences of interest, few of them can detect somatic mutations due to their insufficient specificity. In this study, we proposed a colorimetric DNA detection method by coupling invasive reaction with nicking endonuclease-assisted nanoparticles amplification (IR-NEANA). A target DNA firstly produces many flaps by invasive reaction. Then the flaps are converted to targets of nicking reaction-assisted nanoparticles amplification by ligation reaction to produce the color change of AuNPs, which can be observed by naked eyes. The detection limit of IR-NEANA was determined as 1pM. Most importantly, the specificity of the method is high enough to pick up as low as 1% mutant from a large amount of wild-type DNA backgrounds. The EGFR gene mutated at c.2573 T>G in 9 tissue samples from non-small cell lung cancer patients were successfully detected by using IR-NEANA, suggesting that our proposed method can be used to detect somatic mutations in biological samples.

  18. 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.; Cohen, Seth; Belini, Andrea; Holwerda, Benne W.; Straughn, Amber; Mechtley, Matthew

    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 = 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.

  19. Amplification of enantiomeric excess, mirror-image symmetry breaking and kinetic proofreading in Soai reaction models with different oligomeric orders.

    Science.gov (United States)

    Micheau, Jean-Claude; Coudret, Christophe; Cruz, José-Manuel; Buhse, Thomas

    2012-10-14

    A comprehensive kinetic analysis of three prototypical autocatalytic cycle models based on the absolute asymmetric Soai reaction is presented. The three models, which can give rise to amplification of enantiomeric excess and mirror-image symmetry breaking, vary by their monomeric, dimeric or trimeric order of the assumed catalytic species. Our numerical approach considered the entire chiral combinatorics of the diastereomeric interactions in the models as well as the multiplicity of coupled reversible reactions without applying fast equilibration or quasi-steady state approximations. For the simplest monomeric model, an extensive range of parameters was explored employing a random grid parameter scanning method that revealed the influence of the parameter values on the product distribution, the reaction-time, the attenuation or amplification of enantiomeric excess as well as on the presence or absence of mirror-image symmetry breaking. A symmetry breaking test was imposed on the three models showing that an increase in the catalytic oligomer size from one to three leads to a higher tolerance to poorer chiral recognition between the diastereoisomers and identifies the greater impact of the diastereoisomeric energy difference over an imperfect stereoselectivity in the catalytic step. This robustness is understood as a particular case of so-called kinetic proofreading in asymmetric autocatalysis.

  20. Amplification of plasmid DNA bound on soil colloidal particles and clay minerals by the polymerase chain reaction

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Polymerase chain reaction (PCR) was used to amplify a 600-base pair (bp) sequence of plasmid pGEX-2T DNA bound on soil colloidal particles from Brown soil (Alfisol) and Red soil (Ultisol), and three different minerals (goethite, kaolinite, montmorillonite). DNA bound on soil colloids, kaolinite, and montmorillonite was not amplified when the complexes were used directly but amplification occurred when the soil colloid or kaolinite-DNA complex was diluted, 10- and 20-fold. The montmorillonite-DNA complex required at least 100-fold dilution before amplification could be detected. DNA bound on goethite was amplified irrespective of whether the complex was used directly, or diluted 10- and 20-fold. The amplification of mineral-bound plasmid DNA by PCR is, therefore, markedly influenced by the type and concentration of minerals used. This information is of fundamental importance to soil molecular microbial ecology with particular reference to monitoring the fate of genetically engineered microorganisms and their recombinant DNA in soil environments.

  1. Sensitive electrochemical determination of miRNAs based on a sandwich assay onto magnetic microcarriers and hybridization chain reaction amplification.

    Science.gov (United States)

    Torrente-Rodríguez, R M; Campuzano, S; Montiel, V Ruiz-Valdepeñas; Montoya, J J; Pingarrón, J M

    2016-12-15

    A novel electrochemical approach for determination of miRNAs involving a sandwich hybridization assay onto streptavidin-magnetic beads (Strep-MBs), hybridization chain reaction (HCR) amplification and amperometric detection at disposable screen-printed carbon electrodes is reported. Using miRNA-21 as the target analyte, a dynamic linear range from 0.2 to 5.0nM with a 60pM (1.5fmol in 25μL) detection limit was obtained. The achieved sensitivity is 24-fold higher than a non-HCR amplification approach involving conventional sandwich type assay onto MBs. Moreover, the whole assay time lasted 1h 45min which is remarkably shorter than other reported methodologies. The methodology exhibited full selectivity against other non-complementary miRNAs as well as an acceptable discrimination between homologous miRNA family members. The applicability of this novel approach was demonstrated by determining mature miRNA-21 in total RNA (RNAt) extracted from tumor cells and human tissues.

  2. Signal Amplification by Enzymatic Reaction in an Immunosensor Based on Localized Surface Plasmon Resonance (LSPR

    Directory of Open Access Journals (Sweden)

    Yong-Beom Shin

    2010-03-01

    Full Text Available An enzymatic reaction was employed as a means to enhance the sensitivity of an immunosensor based on localized surface plasmon resonance (LSPR. The reaction occurs after intermolecular binding between an antigen and an antibody on gold nano-island (NI surfaces. For LSPR sensing, the gold NI surface was fabricated on glass substrates using vacuum evaporation and heat treatment. The interferon-g (IFN-g capture antibody was immobilized on the gold NIs, followed by binding of IFN-g to the antibody. Subsequently, a biotinylated antibody and a horseradish peroxidase (HRP conjugated with avidin were simultaneously introduced. A solution of 4-chloro-1-naphthol (4-CN was then used for precipitation; precipitation was the result of the enzymatic reaction catalyzed the HRP on gold NIs. The LSPR spectra were obtained after each binding process. Using this method, the enzyme-catalyzed precipitation reaction on the gold NI surface was found to effectively amplify the change in the signal of the LSPR immunosensor after intermolecular binding.

  3. 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.

  4. A power-efficient thermocycler based on induction heating for DNA amplification by polymerase chain reaction

    Science.gov (United States)

    Pal, Debjani; Venkataraman, V.; Mohan, K. Naga; Chandra, H. Sharat; Natarajan, Vasant

    2004-09-01

    We have built a thermocycler based on the principles of induction heating for polymerase chain reaction (PCR) of target sequences in DNA samples of interest. The cycler has an average heating rate of ˜0.8 °C/s and a cooling rate of ˜0.5 °C/s, and typically takes ˜4 h to complete a 40-cycle PCR protocol. It is power-efficient (˜6 W per reaction tube), micro-processor controlled, and can be adapted for battery operation. Using this instrument, we have successfully amplified a 350 bp segment from a plasmid and SRY, the human sex determining gene, which occurs as a single-copy sequence in genomic DNA of human males. The PCR products from this thermocycler are comparable to those obtained by the use of commercially available machines. Its easy front-end operation, low-power design, portability and low cost makes it suitable for diagnostic field applications of PCR.

  5. A Piezoelectric Immunosensor Based on Agglutination Reaction with Amplification of Silica Nanoparticles

    Institute of Scientific and Technical Information of China (English)

    JIN,Xiao-Yong; JIN,Xue-Fang; DING,Yan-Jun; JIANG,Jian-Hui; SHEN,Guo-Li; Ru-Qin

    2008-01-01

    A simple piezoelectric immunoagglutination assay technique with antibody-modified nanoparticles has been developed for direct quantitative detection of protein. The proposed technique is based on the specific agglutination of goat anti-hlgG-coated silica (or gold) nanoparticles in the presence of human immunoglobulin G (hlgG), which causes a frequency change and is monitored by a piezoelectric device. The antibody modified on the probe surface would combine with antibody-coated nanoparticles in the presence of antigen (hlgG) when the surface agglutination reaction took place, which couples both the mass effect and viscoelastic effect acting on the probe. The results indi-cate that the background interference can be substantially minimized and the probe signal can be observably multi-plied. In addition, the surface of the modified probe and that after combining the complex of immunoagglutination were imaged by scanning electronic microscopy (SEM). Moreover, an optimization of assay medium composition with the addition of poly(ethylene glycol) (PEG) serving as immunoagglutination enhancer and sodium chloride to control the ion-strength was investigated. The frequency responses of the immunoagglutination assay were found to correlate well with the hlgG concentration with a detection limit of 84 ng.mL-1.

  6. Haplotyping using a combination of polymerase chain reaction-single-strand conformational polymorphism analysis and haplotype-specific PCR amplification.

    Science.gov (United States)

    Zhou, Huitong; Li, Shaobin; Liu, Xiu; Wang, Jiqing; Luo, Yuzhu; Hickford, Jon G H

    2014-12-01

    A single nucleotide polymorphism (SNP) may have an impact on phenotype, but it may also be influenced by multiple SNPs within a gene; hence, the haplotype or phase of multiple SNPs needs to be known. Various methods for haplotyping SNPs have been proposed, but a simple and cost-effective method is currently unavailable. Here we describe a haplotyping approach using two simple techniques: polymerase chain reaction-single-strand conformational polymorphism (PCR-SSCP) and haplotype-specific PCR. In this approach, individual regions of a gene are analyzed by PCR-SSCP to identify variation that defines sub-haplotypes, and then extended haplotypes are assembled from the sub-haplotypes either directly or with the additional use of haplotype-specific PCR amplification. We demonstrate the utility of this approach by haplotyping ovine FABP4 across two variable regions that contain seven SNPs and one indel. The simplicity of this approach makes it suitable for large-scale studies and/or diagnostic screening.

  7. A survey of polymerase chain reaction (PCR) amplification studies of unicellular protists using single-cell PCR.

    Science.gov (United States)

    Lynn, Denis H; Pinheiro, Marcel

    2009-01-01

    We surveyed a variety of studies that have used single-cell polymerase chain reaction (SC-PCR) to examine the gene sequences of a diversity of unicellular protists. Representatives of all the Super-Groups of eukaryotes have been subjected to SC-PCR with ciliates and dinoflagellates being most commonly examined. The SC-PCR was carried out either by directly amplifying a single lysed cell or by first extracting DNA and following this with amplification of the DNA extract. Cell lysis methods included heating, freezing, mechanical rupture, and enzyme digestion. Cells fixed or preserved with ethanol, methanol, and Lugol's have also been used successfully. Heminested or seminested PCR might follow the initial PCR, whose products were then directly sequenced or cloned and then sequenced. The methods are not complicated. This should encourage protistologists to use SC-PCR in the description of new or revised taxa, especially rare and unculturable forms, and it should also enable the probing of gene expression in relation to life history stages.

  8. Sensitive and selective amplification of methylated DNA sequences using helper-dependent chain reaction in combination with a methylation-dependent restriction enzymes.

    Science.gov (United States)

    Rand, Keith N; Young, Graeme P; Ho, Thu; Molloy, Peter L

    2013-01-07

    We have developed a novel technique for specific amplification of rare methylated DNA fragments in a high background of unmethylated sequences that avoids the need of bisulphite conversion. The methylation-dependent restriction enzyme GlaI is used to selectively cut methylated DNA. Then targeted fragments are tagged using specially designed 'helper' oligonucleotides that are also used to maintain selection in subsequent amplification cycles in a process called 'helper-dependent chain reaction'. The process uses disabled primers called 'drivers' that can only prime on each cycle if the helpers recognize specific sequences within the target amplicon. In this way, selection for the sequence of interest is maintained throughout the amplification, preventing amplification of unwanted sequences. Here we show how the method can be applied to methylated Septin 9, a promising biomarker for early diagnosis of colorectal cancer. The GlaI digestion and subsequent amplification can all be done in a single tube. A detection sensitivity of 0.1% methylated DNA in a background of unmethylated DNA was achieved, which was similar to the well-established Heavy Methyl method that requires bisulphite-treated DNA.

  9. Amplification refractory mutation system polymerase chain reaction versus optimized polymerase chain reaction restriction-fragment length polymorphism for apolipoprotein E genotyping of majorly depressed patients.

    Science.gov (United States)

    You, Hongmin; Chen, Jin; Zhou, Jingjing; Huang, Hua; Pan, Junxi; Wang, Ziye; Lv, Lin; Zhang, Lujun; Li, Juan; Qin, Bin; Yang, Yongtao; Xie, Peng

    2015-11-01

    Major depressive disorder (MDD) is a prevalent, debilitating mood disorder that has been associated with several genetic polymorphisms. One such polymorphism, namely that of apolipoprotein E (APOE), has three allelic forms (ε2, ε3 and ε4) that encode for six unique isoforms of the APOE protein. A growing number of techniques have been developed for APOE genotyping; however, not all polymerase chain reaction (PCR)‑based genotyping techniques are equally accurate or cost‑effective. In order to find a more accurate and cost‑effective APOE genotyping method for MDD screening in large populations, the present study comparatively evaluated two genotyping methods, amplification refractory mutation system PCR (ARMS‑PCR) and optimized PCR restriction‑fragment length polymorphism (PCR‑RFLP), in blood samples taken from a population of 708 MDD patients. Although either of the two methods were able to detect all six unique APOE genotypes, comparisons of the two methods with Sanger sequencing demonstrated that ARMS‑PCR (94%) was significantly more accurate than optimized PCR‑RFLP (82%). ARMS‑PCR should prove useful in quickly verifying ambiguous results obtained by other APOE genotyping methods and can be cost-effectively performed in the setting of a small laboratory or a population-based screening program.

  10. Sensitive electrochemical detection of telomerase activity using spherical nucleic acids gold nanoparticles triggered mimic-hybridization chain reaction enzyme-free dual signal amplification.

    Science.gov (United States)

    Wang, Wen-Jing; Li, Jing-Jing; Rui, Kai; Gai, Pan-Pan; Zhang, Jian-Rong; Zhu, Jun-Jie

    2015-03-03

    We report an electrochemical sensor for telomerase activity detection based on spherical nucleic acids gold nanoparticles (SNAs AuNPs) triggered mimic-hybridization chain reaction (mimic-HCR) enzyme-free dual signal amplification. In the detection strategy, SNAs AuNPs and two hairpin probes were employed. SNAs AuNPs as the primary amplification element, not only hybridized with the telomeric repeats on the electrode to amplify signal but also initiated the subsequent secondary amplification, mimic-hybridization chain reaction of two hairpin probes. If the cells' extracts were positive for telomerase activity, SNAs AuNPs could be captured on the electrode. The carried initiators could trigger an alternative hybridization reaction of two hairpin probes that yielded nicked double helices. The signal was further amplified enzyme-free by numerous hexaammineruthenium(III) chloride ([Ru(NH3)6](3+), RuHex) inserting into double-helix DNA long chain by electrostatic interaction, each of which could generate an electrochemical signal at appropriate potential. With this method, a detection limit of down to 2 HeLa cells and a dynamic range of 10-10,000 cells were achieved. Telomerase activities of different cell lines were also successfully evaluated.

  11. 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.

  12. Targeted rapid amplification of cDNA ends (T-RACE)--an improved RACE reaction through degradation of non-target sequences.

    Science.gov (United States)

    Bower, Neil I; Johnston, Ian A

    2010-11-01

    Amplification of the 5' ends of cDNA, although simple in theory, can often be difficult to achieve. We describe a novel method for the specific amplification of cDNA ends. An oligo-dT adapter incorporating a dUTP-containing PCR primer primes first-strand cDNA synthesis incorporating dUTP. Using the Cap finder approach, another distinct dUTP containing adapter is added to the 3' end of the newly synthesized cDNA. Second-strand synthesis incorporating dUTP is achieved by PCR, using dUTP-containing primers complimentary to the adapter sequences incorporated in the cDNA ends. The double-stranded cDNA-containing dUTP serves as a universal template for the specific amplification of the 3' or 5' end of any gene. To amplify the ends of cDNA, asymmetric PCR is performed using a single gene-specific primer and standard dNTPs. The asymmetric PCR product is purified and non-target transcripts containing dUTP degraded by Uracil DNA glycosylase, leaving only those transcripts produced during the asymmetric PCR. Subsequent PCR using a nested gene-specific primer and the 3' or 5' T-RACE primer results in specific amplification of cDNA ends. This method can be used to specifically amplify the 3' and 5' ends of numerous cDNAs from a single cDNA synthesis reaction.

  13. Amplification of an invA gene sequence of Salmonella typhimurium by polymerase chain reaction as a specific method of detection of Salmonella.

    Science.gov (United States)

    Rahn, K; De Grandis, S A; Clarke, R C; McEwen, S A; Galán, J E; Ginocchio, C; Curtiss, R; Gyles, C L

    1992-08-01

    Amplification of nucleotide sequences within the invA gene of Salmonella typhimurium was evaluated as a means of detecting Salmonella. A collection of 630 strains of Salmonella comprising over 100 serovars, including the 20 most prevalent serovars isolated from animals and humans in Canada, was examined. Controls consisted of 142 non-Salmonella strains comprising 21 genera of bacteria. Cultures were screened by inoculating a single colony of bacteria directly into a polymerase chain reaction (PCR) mixture which contained a pair of primers specific for the invA gene. The specific PCR product was a 284 bp DNA fragment which was visualized in 2% agarose gels. With the exception of two S. litchfield and two S. senftenberg strains, all Salmonella strains were detected. In contrast, none of the non-Salmonella strains yielded the specific amplification product. Non-specific amplification of a few non-Salmonella strains resulted in a product that was distinctly different in size from the specific 284 bp product. Specificity of amplification was further confirmed by demonstration of hybridization of a 32P-labelled invA gene fragment only to the specific 284 bp product. The detection of 99.4% of Salmonella strains tested and the failure to specifically amplify DNA from non-Salmonella strains confirm that the invA gene contains sequences unique to Salmonella and demonstrate that this gene is a suitable PCR target, with potential diagnostic applications.

  14. Identification of goose (Anser anser) and mule duck (Anasplatyrhynchos x Cairina moschata) foie gras by multiplex polymerase chain reaction amplification of the 5S RDNA gene.

    Science.gov (United States)

    Rodríguez, M A; García, T; González, I; Asensio, L; Fernández, A; Lobo, E; Hernández, P E; Martín, R

    2001-06-01

    Polymerase chain reaction (PCR) amplification of the nuclear 5S rDNA gene has been used for the identification of goose and mule duck foie gras. Two species-specific reverse primers were designed and used in a multiplex reaction, together with a forward universal primer, to amplify specific fragments of the 5S rDNA in each species. The different sizes of the species-specific amplicons, separated by agarose gel electrophoresis, allowed clear identification of goose and mule duck foie gras samples. This genetic marker can be useful for detecting fraudulent substitution of the duck liver for the more expensive goose liver.

  15. The PEAR proposition.

    Science.gov (United States)

    Jahn, R G; Dunne, B J

    2007-01-01

    For more than a quarter century, the Princeton Engineering Anomalies Research (PEAR) laboratory has engaged in a broad range of experiments on consciousness-related physical anomalies and has proposed a corresponding selection of theoretical models that have combined to illuminate the fundamental nature of the provocative phenomena that emerge. Productive pursuit of this topic has inescapably involved a spectrum of political, cultural, personal, and interpersonal factors that are normally not encountered in more conventional scientific scholarship, but have both enriched and complicated the enterprise in many ways. Some of the insights gleaned from the work are objectively specifiable, such as the scale and structural character of the anomalous effects; their relative insensitivity to objective physical correlates, including distance and time; the oscillating sequential patterns of performance they display; the major discrepancies between male and female achievements; and their irregular replicability at all levels of experience. But many others relate to subjective issues, such as the responsiveness of the effects to conscious and unconscious intention and to individual and collective resonance; the relevance of ambience and attitude in their generation; and the importance of intrinsic uncertainty as a source of the anomalies. This blend of empirical features predicates radical excursions of the dedicated models, and hence of the more general scientific paradigms, to allow consciousness and its subjective information processing capacities a proactive role in the establishment of objective reality, with all of the complications of specificity, causality, and reproducibility that entails. The attendant complexities of conceptualization, formulation, and implementation notwithstanding, pragmatic applications of these phenomena in many sectors of public endeavor now can be foreseen.

  16. Revolutions in rapid amplification of cDNA ends: new strategies for polymerase chain reaction cloning of full-length cDNA ends.

    Science.gov (United States)

    Schaefer, B C

    1995-05-20

    Rapid amplification of cDNA ends (RACE) is a polymerase chain reaction (PCR)-based technique which was developed to facilitate the cloning of full-length cDNA 5'- and 3'-ends after a partial cDNA sequence has been obtained by other methods. While RACE can yield complete sequences of cDNA ends in only a few days, the RACE procedure frequently results in the exclusive amplification of truncated cDNA ends, undermining efforts to generate full-length clones. Many investigators have suggested modifications to the RACE protocol to improve the effectiveness of the technique. Based on first-hand experience with RACE, a critical review of numerous published variations of the key steps in the RACE method is presented. Also included is a detailed, effective protocol based on RNA ligase-mediated RACE/reverse ligation-mediated PCR, as well as a demonstration of its utility.

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

    Science.gov (United States)

    Balne, P K; Basu, S; Rath, S; Barik, M R; Sharma, S

    2015-01-01

    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.

  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. 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.

  20. T Oligo-Primed Polymerase Chain Reaction (TOP-PCR): A Robust Method for the Amplification of Minute DNA Fragments in Body Fluids

    Science.gov (United States)

    Nai, Yu-Shin; Chen, Tzu-Han; Huang, Yu-Feng; Midha, Mohit K.; Shiau, Hsin-Chieh; Shen, Chen-Yang; Chen, Chien-Jen; Yu, Alice L.; Chiu, Kuo Ping

    2017-01-01

    Body fluid DNA sequencing is a powerful noninvasive approach for the diagnosis of genetic defects, infectious agents and diseases. The success relies on the quantity and quality of the DNA samples. However, numerous clinical samples are either at low quantity or of poor quality due to various reasons. To overcome these problems, we have developed T oligo-primed polymerase chain reaction (TOP-PCR) for full-length nonselective amplification of minute quantity of DNA fragments. TOP-PCR adopts homogeneous “half adaptor” (HA), generated by annealing P oligo (carrying a phosphate group at the 5′ end) and T oligo (carrying a T-tail at the 3′ end), for efficient ligation to target DNA and subsequent PCR amplification primed by the T oligo alone. Using DNA samples from body fluids, we demonstrate that TOP-PCR recovers minute DNA fragments and maintains the DNA size profile, while enhancing the major molecular populations. Our results also showed that TOP-PCR is a superior method for detecting apoptosis and outperforms the method adopted by Illumina for DNA amplification. PMID:28094343

  1. 76 FR 54075 - Pears Grown in Oregon and Washington; Assessment Rate Decrease for Fresh Pears

    Science.gov (United States)

    2011-08-31

    ... winter pear promotion budget for the 2011-2011 fiscal period was reduced. The major expenditures... by Pear Bureau Northwest, $610,700 for 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...

  2. Extrusion Processing of Cactus Pear

    Directory of Open Access Journals (Sweden)

    Preetam Sarkar

    2011-04-01

    Full Text Available Whole fruit utilization using extrusion technology has received limited attention in the food processing industry. The objective of this study was to investigate the utilization of prickly pear fruit solids in extruded food products. Peeled prickly pear fruits were ground to form a paste. This paste was strained to remove the seeds and then mixed with rice flour in three different solid ratios. The three blends were dried to a moisture level of 13% (w/w basis and ground to form fine flour. These feed mixes were extruded in a twin screw extruder (Clextral EV-25 at a feed rate of 15 kg/h, feed moisture content of 13% (w/w, screw speed of 400 rpm and L/D ratio of 40:1. The temperature profile from feed to die end was maintained as: 25, 30, 40, 50, 60, 70, 80, 100, 120, 140ºC. The extruded products were analyzed for physical and textural properties. Apparent density and breaking strength of the cactus pear extrudates increased from 116.07 to 229.66 kg/m3 and 58.5 to 178.63 kPa, respectively with increase in fruit solid level. However, true density, porosity and radial expansion ratio decreased from 837.89 to 775.84 kg/m3, 86.12 to 70.34% and 12.37 to 6.6, respectively with increase in fruit solid level. This study demonstrated the potential of extrusion processing to utilize peeled cactus pear fruits for production of expanded food products.

  3. 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.

  4. Polymerase reaction without primers throughout for the reconstruction of full-length cDNA from products of rapid amplification of cDNA ends (RACE).

    Science.gov (United States)

    Sunohara, Mitsuhiro; Kawakami, Masanori; Kage, Hidenori; Watanabe, Kousuke; Emoto, Noriko; Nagase, Takahide; Ohishi, Nobuya; Takai, Daiya

    2011-07-01

    Rapid amplification of cDNA ends (RACE) has widely been used to determine both ends of the cDNA from its partial sequence. Conventionally, 5'- and 3'-RACE products were ligated at a restriction site in the overlap region to reconstruct the full-length cDNA; however, reconstruction is difficult if no appropriate restriction enzymes are available. Here, we report a novel method to reconstruct full-length cDNA with DNA polymerase. Instead of usual PCR, chain reactions were avoided and the elongation time was shortened, which enables non-specific products or undesired point mutations to be minimized. We successfully reconstructed and TA-cloned a full-length cDNA of echinoderm microtubule-associated protein-like 4 (EML4)-anaplastic lymphoma kinase (ALK) fusion gene variant 2 from RACE products obtained from a surgically resected lung adenocarcinoma sample. We also evaluated some parameters to provide recommendations for this new method.

  5. 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.

  6. 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.

  7. 山苍子AFLP反应体系的建立及其引物筛选%Primer Screening and AFLP Amplification Reaction System of Litsea cubeba

    Institute of Scientific and Technical Information of China (English)

    田胜平; 汪阳东; 陈益存; 占志勇; 斯林林

    2012-01-01

    The effect of DNA extraction was analyzed by comparing the young leaf, terminal bud and flower of Litsea cubeba. The time of DNA digestion and several key factors affecting the PCR selective amplification such as Mg2 + concentration, dNTPs concentration and the mount of the selective amplification primer were also trialed. An optimized AFLP reaction system of Litsea cubeba was established. The results showed that the high quality genomic DNA as a PCR template could be isolated from the bud tissue; genomic DNA could be digested in a hour by 5 U EcoR I and 5 U Mse I; The optical selection amplification system was 20 (jlL reaction mix containing 1.0 U rTaq polymer-ase, 2.0 μL 10 xPCR buffer, 1.8 μL25 mmol ·L-1MgCl2, 1.4 μL2.5 mmol · LT-1dNTP, 100 ng · μ-1 primer each 1.0 μL. Clear and stable amplification band patterns can be obtained and 10 pairs of AFLP primers with good genetic diversity were selected according to the optimized reaction system. The results will be an effective protocol for further studying the genetic structure and differentiation of Litsea cubeba population.%通过对山苍子幼嫩叶片、顶芽、花蕾3种组织的DNA提取效果分析和对影响酶切及选择性扩增效果的4个主要因素(酶切时间、Mg2+浓度、dNTPs浓度、引物浓度)的比较研究,建立了适合于山苍子AFLP分析的技术体系.结果表明,山苍子的顶芽是较好的DNA提取材料;山苍子基因组DNA经5 UEcoRI和5 UMseI酶切lh即可完全酶切;最佳的选择性扩增体系为20 μL反应体系中含有1.0U rTaq聚合酶、2.0 μL 10×PCR缓冲液、1.8 μL 25mmol·L-1MgCl2、1.4 μL 2.5 mmol·L-1dNTP、100 ng·μL-1引物各1.0 μL.使用该反应体系获得了清晰、稳定的DNA指纹图谱,并筛选出10对多态性较好的AFLP引物组合,为利用AFLP标记技术进一步开展山苍子种群遗传结构、遗传分化等研究奠定了基础.

  8. Amplification of Chloroplast DNA Using the Polymerase Chain Reaction (PCR): A Practical Activity for Secondary School Students

    Science.gov (United States)

    Hamilton, Kenny; Barfoot, Jan; Crawford, Kathleen E.; Simpson, Craig G.; Beaumont, Paul C.; Bownes, Mary

    2006-01-01

    We describe a polymerase chain reaction (PCR) protocol suitable for use in secondary schools and colleges. This PCR protocol can be used to investigate genetic variation between plants. The protocol makes use of primers which are complementary to sequences of nucleotides that are highly conserved across different plant genera. The regions of…

  9. 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.

  10. 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.

  11. DNA Extraction from Formalin-fixed and Paraffin-embedded Tissues by Triton X-100 for Effective Amplification of EGFR Gene by Polymerase Chain Reaction

    Institute of Scientific and Technical Information of China (English)

    WANG Xiao-feng; DU Zhen-wu; WU Mei; ZHANG Yu-cheng; JIANG Yang; ZHANG Gui-zhen

    2012-01-01

    For first-line non-small-cell lung cancer(NSCLC) therapy,detecting mutation status of the epidermal growth factor receptor(EGFR) gene constitutes a prudent test to identify patients who are most likely to benefit from EGFR-tyrosine kinase inhibitor(TKI) therapy.Now,the material for detecting EGFR gene mutation status mainly comes from formalin-fixed and paraffin-embedded(FFPE) tissues.DNA extraction from FFPE and the amplification of EGFR gene by polymerase chain reaction(PCR) are two key steps for detecting EGFR gene mutation.We showed a simple method of DNA extraction from FFPE tissues for the effective amplification of EGFR gene.Extracting DNA from the FFPE tissues of NSCLC patients with 1% Triton X-100(pH=10.0) was performed by heating at 95 C for 30min.Meanwhile,a commercial kit was used to extract DNA from the same FFPE tissues of NSCLC patients for comparison.DNA extracted products were used as template for amplifying the exons 18,.19,20 and 21 of EGFR by PCR for different amplified fragments.Results show that DNA fragment size extracted from FFPE tissues with 1%Triton X was about 250-500 base pairs(bp).However,DNA fragment size extracted from FFPE tissues via commercial kit was about from several hundreds to several thousands bp.The DNA yield extracted from FFPE tissues with 1% Triton X was larger than that via commercial kit.For about 500 bp fragment,four exons of EGFR could not be amplified more efficiently from extracted DNA with 1% Triton X than with commercial kit.However,for about 200 bp fragment.This simple and non-laborious protocol could successfully be used to extract DNA from FFPE tissue for the amplification of EGFR gene by PCR,further screening of EGFR gene mutation and facilitating the molecular analysis of a large number of FFPE tissues from NSCLC patients.

  12. A new morphologically distinct avian malaria parasite that fails detection by established polymerase chain reaction-based protocols for amplification of the cytochrome B gene.

    Science.gov (United States)

    Zehtindjiev, Pavel; Križanauskienė, Asta; Bensch, Staffan; Palinauskas, Vaidas; Asghar, Muhammad; Dimitrov, Dimitar; Scebba, Sergio; Valkiūnas, Gediminas

    2012-06-01

    Plasmodium polymorphum n. sp. (Haemosporida, Plasmodiidae) was found in the skylark, Alauda arvensis (Passeriformes: Alaudidae), during autumnal migration in southern Italy. This organism is illustrated and described based on the morphology of its blood stages. The most distinctive feature of this malaria parasite is the clear preference of its blood stages (trophozoites, meronts, and gametocytes) for immature red blood cells, including erythroblasts. Based on preference of erythrocytic meronts for immature red blood cells, P. polymorphum is most similar to species of the subgenus Huffia . This parasite can be readily distinguished from all other bird malaria parasites, including Plasmodium ( Huffia ) spp., due to preferential development and maturation of its gametocytes in immature red blood cells, a unique character for avian Plasmodium spp. In addition, the margins of nuclei in blood stages of P. polymorphum are markedly smooth and distinct; this is also a distinct diagnostic feature of this parasite. Plasmodium polymorphum has been recorded only in the skylark; it is probably a rare parasite, whose host range and geographical distribution remain unclear. Microscopic examination detected a light infection of Plasmodium relictum (lineage GRW11, parasitemia of mitochondrial cytochrome b gene (cyt b ) of P. polymorphum from the microscopically positive sample by using published and newly designed primers for DNA amplification of avian Plasmodium spp. The light parasitemia of P. relictum was easily detectable using several polymerase chain reaction (PCR)-based assays, but P. polymorphum was undetectable in all applied assays. Quantitative PCR also showed the presence of light parasitemia (0.06%) of the lineage GRW11 in this sample. This supports the conclusion that the morphologically distinct parasite observed along with P. relictum and predominant in the sample is genetically dissimilar from the lineage GRW11 based on cyt b sequence. In samples with co

  13. Sex determination of porcine embryos using a new developed duplex polymerase chain reaction procedure based on the amplification of repetitive sequences.

    Science.gov (United States)

    Torner, Eva; Bussalleu, Eva; Briz, M Dolors; Gutiérrez-Adán, Alfonso; Bonet, Sergi

    2013-01-01

    Polymerase chain reaction (PCR)-based assays have become increasingly prevalent for sexing embryos. The aim of the present study was to develop a suitable duplex PCR procedure based on the amplification of porcine repetitive sequences for sexing porcine tissues, embryos and single cells. Primers were designed targeting the X12696 Y chromosome-specific repeat sequence (SUSYa and SUSYb; sex-related primer sets), the multicopy porcine-specific mitochondrial 12S rRNA gene (SUS12S; control primer set) and the X51555 1 chromosome repeat sequence (SUS1; control primer set). The specificity of the primer sets was established and the technique was optimised by testing combinations of two specific primer sets (SUSYa/SUS12S; SUSYb/SUS12S), different primer concentrations, two sources of DNA polymerase, different melting temperatures and different numbers of amplification cycles using genomic DNA from porcine ovarian and testicular tissue. The optimised SUSYa/SUS12S- and SUSYb/SUS12S-based duplex PCR procedures were applied to porcine in vitro-produced (IVP) blastocysts, cell-stage embryos and oocytes. The SUSYb/SUS12S primer-based procedure successfully sexed porcine single cells and IVP cell-stage embryos (100% efficiency), as well as blastocysts (96.6% accuracy; 96.7% efficiency). This is the first report to demonstrate the applicability of these repetitive sequences for this purpose. In conclusion, the SUSYb/SUS12S primer-based duplex PCR procedure is highly reliable and sensitive for sexing porcine IVP embryos.

  14. The pear violin 梨提琴

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    1.There was a big pear on the ground.“What’sthis?”the little squirrel;asked.地上有个大梨。小松鼠说:“这是什么呀?”:“2.He cut the pear into two halves and ate one.小松鼠把梨对半切开,吃掉了半个。

  15. 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.

  16. 77 FR 72197 - Pears Grown in Oregon and Washington; Assessment Rate Decrease for Processed Pears

    Science.gov (United States)

    2012-12-05

    ... above. FOR FURTHER INFORMATION CONTACT: Teresa Hutchinson or Gary Olson, Northwest Marketing Field... pears may be classified as small entities. There are three pear processing plants in the production area... Federal rules that duplicate, overlap, or conflict with this rule. A small business guide on...

  17. 77 FR 72245 - Pears Grown in Oregon and Washington; Committee Membership Reapportionment for Processed Pears

    Science.gov (United States)

    2012-12-05

    ...; ] DEPARTMENT OF AGRICULTURE Agricultural Marketing Service 7 CFR Part 927 Pears Grown in Oregon and Washington; Committee Membership Reapportionment for Processed Pears AGENCY: Agricultural Marketing Service, USDA... under the Agricultural Marketing Agreement Act of 1937, as amended (7 U.S.C. 601-674),...

  18. Isothermal Amplification of Nucleic Acids.

    Science.gov (United States)

    Zhao, Yongxi; Chen, Feng; Li, Qian; Wang, Lihua; Fan, Chunhai

    2015-11-25

    Isothermal amplification of nucleic acids is a simple process that rapidly and efficiently accumulates nucleic acid sequences at constant temperature. Since the early 1990s, various isothermal amplification techniques have been developed as alternatives to polymerase chain reaction (PCR). These isothermal amplification methods have been used for biosensing targets such as DNA, RNA, cells, proteins, small molecules, and ions. The applications of these techniques for in situ or intracellular bioimaging and sequencing have been amply demonstrated. Amplicons produced by isothermal amplification methods have also been utilized to construct versatile nucleic acid nanomaterials for promising applications in biomedicine, bioimaging, and biosensing. The integration of isothermal amplification into microsystems or portable devices improves nucleic acid-based on-site assays and confers high sensitivity. Single-cell and single-molecule analyses have also been implemented based on integrated microfluidic systems. In this review, we provide a comprehensive overview of the isothermal amplification of nucleic acids encompassing work published in the past two decades. First, different isothermal amplification techniques are classified into three types based on reaction kinetics. Then, we summarize the applications of isothermal amplification in bioanalysis, diagnostics, nanotechnology, materials science, and device integration. Finally, several challenges and perspectives in the field are discussed.

  19. G-quadruplex structures and CpG methylation cause drop-out of the maternal allele in polymerase chain reaction amplification of the imprinted MEST gene promoter.

    Directory of Open Access Journals (Sweden)

    Aaron J Stevens

    Full Text Available We observed apparent non-Mendelian behaviour of alleles when genotyping a region in a CpG island at the 5' end of the maternally imprinted human MEST isoform. This region contains three single nucleotide polymorphisms (SNPs in total linkage disequilibrium, such that only two haplotypes occur in the human population. Only one haplotype was detectable in each subject, never both, despite the use of multiple primers and several genotyping methods. We observed that this region contains motifs capable of forming several G-quadruplex structures. Circular dichroism spectroscopy and native polyacrylamide gel electrophoresis confirmed that at least three G-quadruplexes form in vitro in the presence of potassium ions, and one of these structures has a Tm of greater than 99°C in polymerase chain reaction (PCR buffer. We demonstrate that it is the methylated maternal allele that is always lost during PCR amplification, and that formation of G-quadruplexes and presence of methylated cytosines both contributed to this phenomenon. This observed parent-of-origin specific allelic drop-out has important implications for analysis of imprinted genes in research and diagnostic settings.

  20. Identification of region-specific yeast artificial chromosomes using pools of Alu element-mediated polymerase chain reaction probes labeled via linear amplification

    Energy Technology Data Exchange (ETDEWEB)

    Cole, C.G.; Bobrow, M.; Bentley, D.R.; Dunham, I. (United Medical and Dental Schools of Guy' s and St. Thomas Hospitals, London Bridge, London, England (United Kingdom)); Patel, K.; Shipley, J.; Sheer, D. (Imperial Cancer Research Fund, London (United Kingdom))

    1992-12-01

    The ability to identify large numbers of yeast artificial chromosomes (YACS) specific to any given genomic region rapidly and efficiently enhances both the construction of clone maps and the isolation of region-specific landmarks (e.g., polymorphic markers). The authors describe a method of preparing region-specific single-stranded hybridization probes from Alu element-mediated polymerase chain reaction (Alu-PCR) products of somatic cell hybrids for YAC library screening. Pools of up to 50 cloned Alu-PCR products from an irradiation-reduced hybrid containing 22q11.2-q13.1 were labeled to high specific activity by linear amplification using a single vector primer. The resulting single-stranded probes were extensively competed to remove repetitive sequences, while retaining the full complexity of the probe. Extensive coverage of the region by YACs using multiple probe pools was demonstrated as many YACs were detected more than once. In situ analysis using chosen YACs confirmed that the clones were specific for the region. Thus, this pooled probe approach constitutes a rapid method to identify large numbers of YACs relevant to a large chromosomal region. 29 refs., 4 figs.

  1. Identification of region-specific yeast artificial chromosomes using pools of Alu element-mediated polymerase chain reaction probes labeled via linear amplification.

    Science.gov (United States)

    Cole, C G; Patel, K; Shipley, J; Sheer, D; Bobrow, M; Bentley, D R; Dunham, I

    1992-12-01

    The ability to identify large numbers of yeast artificial chromosomes (YACs) specific to any given genomic region rapidly and efficiently enhances both the construction of clone maps and the isolation of region-specific landmarks (e.g., polymorphic markers). We describe a method of preparing region-specific single-stranded hybridization probes from Alu element-mediated polymerase chain reaction (Alu-PCR) products of somatic cell hybrids for YAC library screening. Pools of up to 50 cloned Alu-PCR products from an irradiation-reduced hybrid containing 22q11.2-q13.1 were labeled to high specific activity by linear amplification using a single vector primer. The resulting single-stranded probes were extensively competed to remove repetitive sequences, while retaining the full complexity of the probe. Extensive coverage of the region by YACs using multiple probe pools was demonstrated as many YACs were detected more than once. In situ analysis using chosen YACs confirmed that the clones were specific for the region. Thus, this pooled probe approach constitutes a rapid method to identify large numbers of YACs relevant to a large chromosomal region.

  2. Molecular cloning of a cysteine proteinase cDNA from adult Ancylostoma ceylanicum by the method of rapid amplification of cDNA ends using polymerase chain reaction.

    Science.gov (United States)

    Mieszczanek, J; Kofta, W; Wedrychowicz, H

    2000-12-01

    The hookworm Ancylostoma ceylanicum is a parasite of great importance in human and veterinary medicine. The most promising vaccination trials against hookworm infections are based on antigens belonging to the proteinase family. The aim of the present research was to isolate a cysteine proteinase gene from A. ceylanicum. This was achieved by rapid amplification of cDNA ends using polymerase chain reaction (RACE-PCR). A set of consensus oligonucleotide primers was designed to anneal to the conserved coding regions of cysteine proteinase. The PCR products were cloned and sequenced. The novel sequence displayed a high degree of homology with genes of cysteine proteinases known from other hookworm species. In the coding region the nucleotide identity with accp-1, the cysteine proteinase gene of A. caninum, reaches 84.3%. Analysis of the expression of acey-1. the cysteine proteinase gene of A. ceylanicum, suggests that it is produced exclusively in the gland cells of either adult worms or blood-feeding stages of A. ceylanicum.

  3. Quantitative real-time polymerase chain reaction is an alternative method for the detection of HER-2 amplification in formalin-fixed paraffin-embedded breast cancer samples.

    Science.gov (United States)

    Pu, Tianjie; Guo, Peng; Qiu, Yan; Chen, Shinan; Yang, Libo; Sun, Linyong; Ye, Feng; Bu, Hong

    2015-01-01

    Fluorescent in situ hybridization (FISH) and immunohistochemistry (IHC) are the most common methods that are used to quantify HER-2 gene and protein levels, respectively, in human breast cancer. However, due to bad sample quality, some samples are unable to be subjected to a FISH assay. We evaluated 71 formalin-fixed paraffin-embedded (FFPE) breast carcinoma specimens by quantitative real-time polymerase chain reaction (qPCR), IHC, and FISH. We also performed qPCR and FISH assays on delayed formalin-fixed (DDF) samples. The qPCR results were in complete concordance with the results of IHC and FISH. In regards to the DDF samples, the HER-2 fluorescent signal seemed decayed compared with that of the DDF samples after 1 h. However, the qPCR method still works well up to 12 hours. Our results indicated that qPCR was obviously superior to FISH in cases that were not fixed in a reasonable amount of time. However, qPCR can be an alternative method by which to perform HER2 amplification assays in breast cancer.

  4. Amplification of papillomaviral DNA sequences from a high proportion of feline cutaneous in situ and invasive squamous cell carcinomas using a nested polymerase chain reaction.

    Science.gov (United States)

    Munday, John S; Kiupel, Matti; French, Adrienne F; Howe, Laryssa

    2008-10-01

    Squamous cell carcinomas (SCCs) are common skin tumours of cats. Previous studies have suggested that papillomaviral (PV) DNA is detectible within some feline SCCs. A PV DNA sequence has been previously amplified from five feline bowenoid in situ carcinomas (BISCs). Primers specific for this sequence were used in a nested polymerase chain reaction to compare PV detection rates in SCCs to rates within non-SCC skin lesions. Papillomaviral DNA was amplified from 20 of 20 BISC, 17 of 20 invasive SCC and 3 of 17 non-SCC controls. The rate of PV amplification from feline cutaneous SCCs was significantly higher than from non-SCC lesions. These results confirm that feline cutaneous SCCs are associated with PV infection. In humans, there is evidence that PVs promote SCC development within sun-exposed skin. The demonstrated association between PVs and feline cutaneous SCCs suggests, but does not prove, that PVs may also promote feline SCC development. If PVs are oncogenic in cats, prevention of PV infection may reduce feline cutaneous SCC development. To the authors' knowledge, this is the first time that PV DNA has been amplified from a non-SCC sample of feline skin.

  5. 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.

  6. Enzyme-free fluorescent biosensor for the detection of DNA based on core-shell Fe3O4 polydopamine nanoparticles and hybridization chain reaction amplification.

    Science.gov (United States)

    Li, Na; Hao, Xia; Kang, Bei Hua; Xu, Zhen; Shi, Yan; Li, Nian Bing; Luo, Hong Qun

    2016-03-15

    A novel, highly sensitive assay for quantitative determination of DNA is developed based on hybridization chain reaction (HCR) amplification and the separation via core-shell Fe3O4 polydopamine nanoparticles (Fe3O4@PDA NPs). In this assay, two hairpin probes are designed, one of which is labeled with a 6-carboxyfluorescein (FAM). Without target DNA, auxiliary hairpin probes are stable in solution. However, when target DNA is present, the HCR between the two hairpins is triggered. The HCR products have sticky ends of 24 nt, which are much longer than the length of sticky ends of auxiliary hairpins (6 nt) and make the adsorption much easier by Fe3O4@PDA NPs. With the addition of Fe3O4@PDA NPs, HCR products could be adsorbed because of the strong interaction between their sticky ends and Fe3O4@PDA NPs. As a result, supernatant of the solution with target DNA emits weak fluorescence after separation by magnet, which is much lower than that of the blank solution. The detection limit of the proposed method is as low as 0.05 nM. And the sensing method exhibits high selectivity for the determination between perfectly complementary sequence and target with single base-pair mismatch. Importantly, the application of the sensor for DNA detection in human serum shows that the proposed method works well for biological samples.

  7. RNA Amplification Protocol Leads to Biased Polymerase Chain Reaction Results Especially for Low-Copy Transcripts of Human Bone Marrow-Derived Stromal Cells.

    Directory of Open Access Journals (Sweden)

    Carolin Coenen

    Full Text Available The amplification of RNA is becoming increasingly important, as often only limited amounts of cells are available for gene expression analysis. In this study, the gene expression profile of the 39 human homeobox (HOX genes was analyzed in bone marrow-derived multipotent stromal cells (BM-MSCs by reverse transcription (RT- and quantitative polymerase chain reaction (qPCR. For further unlimited gene expression analysis, Whole Transcriptome Amplification (WTA was used to amplify RNA from human BM-MSCs. However, WTA led to biased RT- and qPCR results, and even non-detectability of HOX transcripts compared to non-amplified BM-MSC samples which instead revealed transcription. It is important to note that the same RNA of the respective human BM-MSC line was used for normal cDNA synthesis by standard reverse transcription (non-amplified RT samples and for cDNA synthesis by WTA (amplified WTA samples. On this account, the different RT- and qPCR results were unexpected applying WTA. The significantly reduced detection of HOX transcripts after WTA has been demonstrated for numerous BM-MSC lines (n = 26 by RT-PCR analysis. Furthermore, undetectable HOX transcripts meaning HOX transcripts of human BM-MSCs that were detected after normal cDNA synthesis, but were no longer detectable after WTA, were consistently observed by qPCR analysis. Finally, qPCR experiments revealed a possible explanation for the differences between amplified and non-amplified BM-MSC samples: an inverse correlation between the biased qPCR results and the low expression level of the respective HOX gene. The PCR analysis of high-copy transcripts like GAPDH or RPL13A revealed unchanged qPCR results after WTA compared to corresponding non-amplified BM-MSC samples. In contrast, WTA led to biased qPCR results for medium-copy HOX transcripts, and even non-detectability of low-copy HOX transcripts of human BM-MSCs resulting in false negative RT- and qPCR data applying WTA.

  8. Molecular identification of mealybugs (Hemiptera: Pseudococcidae) found on Korean pears.

    Science.gov (United States)

    Park, Doo-Sang; Leem, Yu Jin; Hahn, Kyu-Woong; Suh, Soo-Jung; Hong, Ki-Jeong; Oh, Hyun-Woo

    2010-02-01

    Mealybugs are under a strict regulation at foreign trades of agricultural products because they are one of the most economically damaging groups of insects on food crops and ornamental plants. However, the absence of morphological characteristics enabling the discrimination of early life stages often cause a significant delay or rejection of a shipment when infested fruit is discovered, causing significant economic loss. A polymerase chain reaction-based method for species identification was developed for six mealybug species known to infest Korean pears including two regulated insects, Planococcus kraunhiae (Kuwana) and Crisicoccus matsumotoi (Siraiwa). Six sets of species-specific primers were designed based on the sequence comparison of the internal transcribed spacer 1 and 2 regions. Efficiency tests against 29 mealybug samples showed that this method could effectively discriminate different mealybug species regardless of their developmental stages. Blind tests against 11 field collected mealybug nymph samples indicated that a single polymerase chain reaction is enough to discriminate unidentified mealybugs collected on Korean pears. This new method will facilitate trade and export requirements, as well as identify the species at any stage of mealybug intercepted.

  9. Pear quality characteristics by Vis / NIR spectroscopy.

    Science.gov (United States)

    Machado, Nicácia P; Fachinello, José C; Galarça, Simone P; Betemps, Débora L; Pasa, Mateus S; Schmitz, Juliano D

    2012-09-01

    Recently, non-destructive techniques such as the Vis / NIR spectroscopy have been used to evaluate the characteristics of maturation and quality of pears. The study aims to validate the readings by the Vis / NIR spectroscopy as a non-destructive way to assess the qualitative characteristics of pear cultivars 'Williams', 'Packams' and 'Carrick', produced according to Brazilian conditions. The experiment was conducted at the Pelotas Federal University, UFPel, in Pelotas / RS, and the instrument used to measure the fruit quality in a non-destructive way was the NIR- Case spectrophotometer (SACMI, Imola, Italy). To determine pears' soluble solids (SS) and pulp firmness (PF), it was established calibration equations for each variety studied, done from the evaluations obtained by a non-destructive method (NIR-Case) and a destructive method. Further on, it was tested the performance of these readings by linear regressions. The results were significant for the soluble solids parameter obtained by the Vis / NIR spectroscopy; however, it did not achieve satisfactory results for the pear pulp firmness of these cultivars. It is concluded that the Vis / NIR spectroscopy, using linear regression, allows providing reliable estimates of pears' quality levels, especially for soluble solids.

  10. 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...

  11. 77 FR 75007 - Importation of Sand Pears From China

    Science.gov (United States)

    2012-12-19

    ... Health Inspection Service 7 CFR Part 319 RIN 0579-AD42 Importation of Sand Pears From China AGENCY... and vegetables regulations to allow the importation of sand pears (Pyrus pyrifolia) from China into the United States. As a condition of entry, sand pears from areas in China in which the Oriental...

  12. Plains Prickly Pear Cactus Response to Fire and Fuel Loads

    Science.gov (United States)

    Management of prickly pear on rangelands has lead to numerous studies aimed at understanding prickly pear response to various natural and human induced treatments. Information is lacking on Plains prickly pear response to varied fuel loads. Pads of clones from three soil types (claypan, gravel, si...

  13. Rapid quantification of Escherichia coli in food and media using bacteriophage T7 amplification and liquid chromatography-multiple reaction monitoring tandem mass spectrometry.

    Science.gov (United States)

    Banu, Mazlina; Ng, Daniel; Zheng, Lu; Goh, Lin-Tang; Bi, Xuezhi; Ow, Dave Siak-Wei

    2014-12-20

    Conventional microbiological assays have been a valuable tool for specific enumeration of indicative bacteria of relevance to food and public health, but these culture-based methods are time-consuming and require tedious biochemical and morphological identification. In this work, we exploit the ability of bacteriophage T7 to specifically infect Escherichia coli and amplify nearly a 100-fold in 1–2 h. Bacteriophage amplification is integrated with liquid chromatography-multiple reaction monitoring tandem mass spectrometry (LC-MRM–MS/MS) for quantitation of phage-specific peptides. Heavy isotopic 15N labeled T7 is introduced as the inoculum phage and internal standard. Quantification is performed by determining the ratio of phage-specific peptides over the internal standard which value is proportional to E. coli numbers. A broad dynamic range of 6-log orders ranging from 3.0 × 10(3) to 3.0 × 10(9) CFU/ml is attained in LB, while between 4.1 × 10(4)–2.7 × 10(9) CFU/ml and 1.9 × 10(3)–3.0 × 10(7) CFU/ml was enumerated respectively in coconut water and apple juice. With this method, viable E. coli are quantified in 4 h with a detection limit of 3.0 × 10(3) CFU/ml, 4.1 × 10(4) CFU/ml and 1.9 × 10(3) CFU/ml in LB, coconut water and apple juice, respectively. This method has potential as a rapid tool for detection of fecal contamination during food bioprocessing and distribution to safeguard public health.

  14. DNA-based hybridization chain reaction and biotin-streptavidin signal amplification for sensitive detection of Escherichia coli O157:H7 through ELISA.

    Science.gov (United States)

    Guo, Qi; Han, Jiao-Jiao; Shan, Shan; Liu, Dao-Feng; Wu, Song-Song; Xiong, Yong-Hua; Lai, Wei-Hua

    2016-12-15

    This study reported on a novel sandwich enzyme linked immunosorbent assay (ELISA) for the sensitive determination of Escherichia coli O157:H7 (E. coli O157:H7) by using DNA-based hybridization chain reaction (HCR) and biotin-streptavidin signal amplification. The anti-E. coli O157:H7 polyclonal antibody (pAb) was immobilized in the ELISA wells. The anti-E. coli O157:H7 monoclonal antibody (mAb) and initiator strand (DNA1) were labeled on gold nanoparticle (AuNP) to form a mAb-AuNP-DNA1 complex. In the presence of the target E. coli O157:H7, the sandwiched immunocomplex, which is pAb-E. coli O157:H7-mAb-AuNP-DNA1, could be formed. Two types of biotinylated hairpin were subsequently added in the ELISA well. A nicked double-stranded DNA (dsDNA) that contained abundant biotins was formed after HCR. Detection was performed after adding horseradish peroxidase-streptavidin and substrate/chromogen solution. Under optimal conditions, E. coli O157:H7 could be detected in the range of 5×10(2) CFU/mL to 1×10(7) CFU/mL; the limit of detection was 1.08×10(2) CFU/mL in pure culture. The LOD of the novel ELISA was 185 times lower than that of traditional ELISA. The proposed method is considerably specific and can be applied in the detection of whole milk samples inoculated with E. coli O157:H7. The coefficient of variation of in pure culture and in whole milk was 0.99-5.88% and 0.76-5.38%, respectively. This method offers a promising application in the detection of low concentrations of food-borne pathogens.

  15. Paper analytical devices for dynamic evaluation of cell surface N-glycan expression via a bimodal biosensor based on multibranched hybridization chain reaction amplification.

    Science.gov (United States)

    Liang, Linlin; Lan, Feifei; Li, Li; Ge, Shenguang; Yu, Jinghua; Ren, Na; Liu, Haiyun; Yan, Mei

    2016-12-15

    A novel colorimetric/fluorescence bimodal lab-on-paper cyto-device was fabricated based on concanavalin A (Con A)-integrating multibranched hybridization chain reaction (mHCR). The product of mHCR was modified PtCu nanochains (colorimetric signal label) and graphene quantum dot (fluorescence signal label) for in situ and dynamically evaluating cell surface N-glycan expression. In this strategy, preliminary detection was carried out through colorimetric method, if needed, then the fluorescence method was applied for a precise determination. Au-Ag-paper devices increased the surface areas and active sites for immobilizing larger amount of aptamers, and then specifically and efficiently captured more cancer cells. Moreover, it could effectively reduce the paper background fluorescence. Due to the specific recognition of Con A with mannose and the effective signal amplification of mHCR, the proposed strategy exhibited excellent high sensitivity for the cytosensing of MCF-7 cells ranging from 100 to 1.0×10(7) and 80-5.0×10(7) cellsmL(-1) with the detection limit of 33 and 26 cellsmL(-1) for colorimetric and fluorescence, respectively. More importantly, this strategy was successfully applied to dynamically monitor cell-surface multi-glycans expression on living cells under external stimuli of inhibitors as well as for N-glycan expression inhibitor screening. These results implied that this biosensor has potential in studying complex native glycan-related biological processes and elucidating the N-glycan-related diseases in biological and physiological processes.

  16. Advances in Japanese pear breeding in Japan

    OpenAIRE

    Saito, Toshihiro

    2016-01-01

    The Japanese pear (Pyrus pyrifolia Nakai) is one of the most widely grown fruit trees in Japan, and it has been used throughout Japan’s history. The commercial production of pears increased rapidly with the successive discoveries of the chance seedling cultivars ‘Chojuro’ and ‘Nijisseiki’ around 1890, and the development of new cultivars has continued since 1915. The late-maturing, leading cultivars ‘Niitaka’ and ‘Shinko’ were released during the initial breeding stage. Furthermore, systemati...

  17. Dynamics and Control of DNA Sequence Amplification

    CERN Document Server

    Marimuthu, Karthikeyan

    2014-01-01

    DNA amplification is the process of replication of a specified DNA sequence \\emph{in vitro} through time-dependent manipulation of its external environment. A theoretical framework for determination of the optimal dynamic operating conditions of DNA amplification reactions, for any specified amplification objective, is presented based on first-principles biophysical modeling and control theory. Amplification of DNA is formulated as a problem in control theory with optimal solutions that can differ considerably from strategies typically used in practice. Using the Polymerase Chain Reaction (PCR) as an example, sequence-dependent biophysical models for DNA amplification are cast as control systems, wherein the dynamics of the reaction are controlled by a manipulated input variable. Using these control systems, we demonstrate that there exists an optimal temperature cycling strategy for geometric amplification of any DNA sequence and formulate optimal control problems that can be used to derive the optimal tempe...

  18. Estimation of resistance pear cultivars to Erwinia amylovora using artificial immature pear fruits method

    OpenAIRE

    Gavrilović, Veljko; Stanisavljević, Rade; Stošić, Stefan; Stevanović, Miloš; Aleksić, Goran; Stajić, Milica; Dolovac, Nenad

    2014-01-01

    Susceptibility of different pear cultivars to Erwinia amaylovora by artificial inoculated immature pear fruits are shown in this article. According obtained results significant differences among cultivars are confirmed and they could be divided in four groups. Most susceptibly cultivars were Santa Marija. Second group includes Williams, Morettini, Carmen, Hardenpont. As most resistant shown to be Magness, Turandot and two local varietyies Karamanka, as well as another unknown local cultivar. ...

  19. Apple and pear rootstock research in Lithuania

    OpenAIRE

    Kviklys, Darius

    2006-01-01

    The paper presents ongoing apple and pear rootstock trials at the Lithuanian Institute of Horticulture. Rootstock research projects are established in following directions: rootstock and location interaction (Baltic fruit rootstock studies where Byelorussian, Estonian, Latvian, Lithuanian and Polish research institutions are involved); budding high effect on rootstock performance; interstock trials; rootstock effect on fruit quality, ripening time and fruit storage; rootstock and tree trainin...

  20. Dynamics and control of DNA sequence amplification

    Energy Technology Data Exchange (ETDEWEB)

    Marimuthu, Karthikeyan [Department of Chemical Engineering and Center for Advanced Process Decision-Making, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213 (United States); Chakrabarti, Raj, E-mail: raj@pmc-group.com, E-mail: rajc@andrew.cmu.edu [Department of Chemical Engineering and Center for Advanced Process Decision-Making, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213 (United States); Division of Fundamental Research, PMC Advanced Technology, Mount Laurel, New Jersey 08054 (United States)

    2014-10-28

    DNA amplification is the process of replication of a specified DNA sequence in vitro through time-dependent manipulation of its external environment. A theoretical framework for determination of the optimal dynamic operating conditions of DNA amplification reactions, for any specified amplification objective, is presented based on first-principles biophysical modeling and control theory. Amplification of DNA is formulated as a problem in control theory with optimal solutions that can differ considerably from strategies typically used in practice. Using the Polymerase Chain Reaction as an example, sequence-dependent biophysical models for DNA amplification are cast as control systems, wherein the dynamics of the reaction are controlled by a manipulated input variable. Using these control systems, we demonstrate that there exists an optimal temperature cycling strategy for geometric amplification of any DNA sequence and formulate optimal control problems that can be used to derive the optimal temperature profile. Strategies for the optimal synthesis of the DNA amplification control trajectory are proposed. Analogous methods can be used to formulate control problems for more advanced amplification objectives corresponding to the design of new types of DNA amplification reactions.

  1. Oxidative stress and senescence-like status of pear calli co-cultured on suspensions of incompatible quince microcalli.

    Science.gov (United States)

    Nocito, Fabio F; Espen, Luca; Fedeli, Chiara; Lancilli, Clarissa; Musacchi, Stefano; Serra, Sara; Sansavini, Silviero; Cocucci, Maurizio; Sacchi, Gian Attilio

    2010-04-01

    This work presents a simple in vitro system to study physiological, biochemical and molecular changes occurring in a pear callus (Pyrus communis L., cv. Beurré Bosc) grown in close proximity to spatially separated undifferentiated homologous (pear) or heterologous (quince; Cydonia oblonga Mill., East Malling clone C) cells in its neighboring environment. After a 7-day co-culture period, the presence of heterologous cells produced negative effects on the pear callus, whose relative weight increase and adenylate energy charge decreased by 30 and 24%, respectively. Such behavior was associated with a higher O(2) consumption rate (+125%) which did not seem to be coupled to adenosine triphosphate synthesis. Analyses of alternative oxidase and enzymatic activities involved in reactive oxygen species (ROS) detoxification strongly suggested that the higher O(2) consumption rate, measured in the pear callus grown in the heterologous combination, may probably be ascribed to extra-respiratory activities. These, in turn, might contribute to generate metabolic scenarios where ROS-induced oxidative stresses may have the upper hand. The increase in the levels of 2-thiobarbituric acid reactive metabolites, considered as diagnostic indicators of ROS-induced lipid peroxidation, seemed to confirm this hypothesis. Moreover, reverse transcription polymerase chain reaction analysis revealed that the expression levels of a few senescence-associated genes were higher in the pear callus grown in the heterologous combination than in the homologous one. Taken as a whole, physiological and molecular data strongly suggest that undifferentiated cells belonging to a pear graft-incompatible quince clone may induce an early senescence-like status in a closely co-cultured pear callus.

  2. Targeted rapid amplification of cDNA ends (T-RACE)—an improved RACE reaction through degradation of non-target sequences

    OpenAIRE

    Neil I Bower; Johnston, Ian A

    2010-01-01

    Amplification of the 5' ends of cDNA, although simple in theory, can often be difficult to achieve. We describe a novel method for the specific amplification of cDNA ends. An oligo-dT adapter incorporating a dUTP-containing PCR primer primes first-strand cDNA synthesis incorporating dUTP. Using the Cap finder approach, another distinct dUTP containing adapter is added to the 3' end of the newly synthesized cDNA. Second-strand synthesis incorporating dUTP is achieved by PCR, using dUTP-contain...

  3. Variations on the Pear Tree Experiment : different variables, new results?

    OpenAIRE

    Igareda, Paula; Matamala, Anna

    2012-01-01

    Inspired by the Pear Stories Project, the Pear Tree Project has investigated how different cultures and languages describe the same film in order to apply its findings to audio description (AD). Participants from different countries were asked to “write down what they saw” in a controlled setting. This article proposes an alternative experiment, also based on the original Pear Stories Project, which aims to shed light on two issues: how different describer profiles (translation students with ...

  4. Revealed Comparative Advantage and Competitiveness in Pear

    Directory of Open Access Journals (Sweden)

    Jaime de Pablo Valenciano

    2012-06-01

    Full Text Available This article focuses on the study of the comparative advantages and competitiveness in the global pear market. First, it will outline a clear distinction between these two concepts, followed by analysis. This paper provides a new index of competitiveness developed by our research based on the insights offered by a wide range of studies on this subject. The aim is to achieve a new line of analysis to improve and expand the possibilities of present day studies.

  5. Quantitative shearing linear amplification polymerase chain reaction: an improved method for quantifying lentiviral vector insertion sites in transplanted hematopoietic cell systems.

    Science.gov (United States)

    Zhou, Sheng; Bonner, Melissa A; Wang, Yong-Dong; Rapp, Samuel; De Ravin, Suk See; Malech, Harry L; Sorrentino, Brian P

    2015-02-01

    In gene therapy trials targeting blood disorders, it is important to detect dominance of transduced hematopoietic stem cell (HSC) clones arising from vector insertion site (VIS) effects. Current methods for VIS analysis often do not have defined levels of quantitative accuracy and therefore can fail to detect early clonal dominance. We have developed a rapid and inexpensive method for measuring clone size based on random shearing of genomic DNA, minimal exponential PCR amplification, and shear site counts as a quantitative endpoint. This quantitative shearing linear amplification PCR (qsLAM PCR) assay utilizes an internal control sample containing 19 lentiviral insertion sites per cell that is mixed with polyclonal samples derived from transduced human CD34+ cells. Samples were analyzed from transplanted pigtail macaques and from a participant in our X-linked severe combined immunodeficiency (XSCID) lentiviral vector trial and yielded controlled and quantitative results in all cases. One case of early clonal dominance was detected in a monkey transplanted with limiting numbers of transduced HSCs, while the clinical samples from the XSCID trial participant showed highly diverse clonal representation. These studies demonstrate that qsLAM PCR is a facile and quantitative assay for measuring clonal repertoires in subjects enrolled in human gene therapy trials using lentiviral-transduced HSCs.

  6. 7 CFR 917.121 - Changes in nomination of Pear Commodity Committee members.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false Changes in nomination of Pear Commodity Committee... Changes in nomination of Pear Commodity Committee members. Nominations for membership on the Pear Commodity Committee shall be made by the growers of pears in the respective representation areas as...

  7. First Report of Neonectria candida Causing Postharvest Decay on ‘Conference’ Pears in the Netherlands

    NARCIS (Netherlands)

    Wenneker, M.; Pham, K.T.K.; Lemmers, M.E.C.; Boer, De F.A.; Lans, Van Der A.M.; Leeuwen, Van P.J.; Hollinger, T.C.; Thomma, B.P.H.J.

    2016-01-01

    Pear (Pyrus communis) is an important fruit crop in the Netherlands, with a total production of 349,000 tons in 2014, and ‘Conference’ is the main pear cultivar that occupies 75% of the total pear production area. In the Netherlands, pears are kept in controlled atmosphere cold storage up to 11 mont

  8. Aroma-Active Compounds in Bartlett Pears and Their Changes during the Manufacturing Process of Bartlett Pear Brandy.

    Science.gov (United States)

    Zierer, Bianca; Schieberle, Peter; Granvogl, Michael

    2016-12-21

    Application of aroma extract dilution analysis to Bartlett pears and the fermented mash produced thereof revealed 24 and 34 aroma-active compounds in the flavor dilution (FD) factor range between 8 and 8192. Twenty-eight compounds, which have not been described before in Bartlett pears or in fermented pear mash, were identified. While ethyl (E,Z)-2,4-decadienoate (pear-like, metallic odor impression), hexyl acetate (green, fruity), and acetic acid (vinegar-like) showed the highest concentrations in Bartlett pears, ethanol (ethanolic), acetic acid, 3-methyl-1-butanol (malty), 1-hexanol (grassy, marzipan-like), (S)-2- and 3-methylbutanoic acid (sweaty), and 2-phenylethanol (flowery, honey-like) were present at the highest amounts in the fermented mash. The key aroma compounds were quantitated in each pear brandy production step (pears, fermented mash, distillate, and aged distillate) by stable isotope dilution analysis showing a clear influence of each step on the overall aroma of the spirit and, consequently, revealing clearly changing concentrations (e.g., of ethyl (S)-2-methylbutanoate, (E)-β-damascenone, ethyl (E,Z)-2,4-decadienoate, and ethyl (E,E)-2,4-decadienoate) and different aroma perceptions during the manufacturing process. In addition, the concentrations of the so-called "pear esters" ethyl (E,Z)-2,4-decadienoate and ethyl (E,E)-2,4-decadienoate were determined in 6 different pear varieties (Abate Fetel, Anjou, Bartlett, Forelle, Kaiser Alexander, and Packham's Triumph) clearly demonstrating the aroma potential of the variety Bartlett, which is mostly used for brandy production due to the high amounts of both esters eliciting a typical pear-like odor impression.

  9. Advances in Japanese pear breeding in Japan.

    Science.gov (United States)

    Saito, Toshihiro

    2016-01-01

    The Japanese pear (Pyrus pyrifolia Nakai) is one of the most widely grown fruit trees in Japan, and it has been used throughout Japan's history. The commercial production of pears increased rapidly with the successive discoveries of the chance seedling cultivars 'Chojuro' and 'Nijisseiki' around 1890, and the development of new cultivars has continued since 1915. The late-maturing, leading cultivars 'Niitaka' and 'Shinko' were released during the initial breeding stage. Furthermore, systematic breeding by the Horticultural Research Station (currently, NARO Institute of Fruit Tree Science, National Agriculture and Food Research Organization (NIFTS)) began in 1935, which mainly aimed to improve fruit quality by focusing on flesh texture and black spot disease resistance. To date, 22 cultivars have been released, including 'Kosui', 'Hosui', and 'Akizuki', which are current leading cultivars from the breeding program. Four induced mutant cultivars induced by gamma irradiation, which exhibit some resistance to black spot disease, were released from the Institute of Radiation Breeding. Among these cultivars, 'Gold Nijisseiki' has become a leading cultivar. Moreover, 'Nansui' from the Nagano prefectural institute breeding program was released, and it has also become a leading cultivar. Current breeding objectives at NIFTS mainly combine superior fruit quality with traits related to labor and cost reduction, multiple disease resistance, or self-compatibility. Regarding future breeding, marker-assisted selection for each trait, QTL analyses, genome-wide association studies, and genomic selection analyses are currently in progress.

  10. Pear quality characteristics by Vis / NIR spectroscopy

    Directory of Open Access Journals (Sweden)

    Nicácia P. Machado

    2012-09-01

    Full Text Available Recently, non-destructive techniques such as the Vis / NIR spectroscopy have been used to evaluate the characteristics of maturation and quality of pears. The study aims to validate the readings by the Vis / NIR spectroscopy as a non-destructive way to assess the qualitative characteristics of pear cultivars 'Williams', 'Packams' and 'Carrick', produced according to Brazilian conditions. The experiment was conducted at the Pelotas Federal University, UFPel, in Pelotas / RS, and the instrument used to measure the fruit quality in a non-destructive way was the NIR- Case spectrophotometer (SACMI, Imola, Italy. To determine pears' soluble solids (SS and pulp firmness (PF, it was established calibration equations for each variety studied, done from the evaluations obtained by a non-destructive method (NIR-Case and a destructive method. Further on, it was tested the performance of these readings by linear regressions. The results were significant for the soluble solids parameter obtained by the Vis / NIR spectroscopy; however, it did not achieve satisfactory results for the pear pulp firmness of these cultivars. It is concluded that the Vis / NIR spectroscopy, using linear regression, allows providing reliable estimates of pears' quality levels, especially for soluble solids.Recentemente, técnicas não destrutivas como a espectroscopia Vis/NIR têm sido utilizadas para avaliar as características de maturação e qualidade das peras. O trabalho tem como objetivo validar as leituras por espectroscopia Vis/NIR, como forma não destrutiva de avaliar as características qualitativas em peras das cultivares Williams, Packams e Carrick produzidas em condições brasileiras. O experimento foi realizado na Universidade Federal de Pelotas, UFPel, Pelotas/RS e o instrumento utilizado para determinar a qualidade dos frutos de forma não destrutiva foi o espectrofotômetro NIR-Case (SACMI, Imola, Itália. Para a determinação de sólidos solúveis (SS e firmeza

  11. Utility of Loop-mediated Isothermal Amplification Assay, Polymerase Chain Reaction, and ELISA for Diagnosis of Leptospirosis in South Indian Patients

    Science.gov (United States)

    Sengupta, Mallika; Prabhakar, Abhilash Kundavaram Paul; Satyendra, Sowmya; Thambu, David; Abraham, Ooriapadickal Cherian; Balaji, Veeraraghavan; Chen, Hua-Wei; Chao, Chien-Chung; Ching, Wei-Mei; Prakash, John Antony Jude

    2017-01-01

    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.

  12. 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.

  13. Direct polymerase chain reaction amplification of formalin-fixed, paraffin-wax-embedded tissue after automated sequential laser microdissection and pressure catapulting.

    Science.gov (United States)

    O'Kane, S L; Garimella, V; Sivarajasingham, N; Drew, P J; Cawkwell, L

    2007-02-01

    A robust method to facilitate rapid laser microdissection and pressure catapulting (LMPC) coupled with direct polymerase chain reaction (dPCR) to eliminate the need for extraction of DNA before a PCR-based assay is described. This sequential LMPC-dPCR method is rapid and decreases the number of processing steps, reducing the chance of tissue loss and contamination.

  14. 76 FR 78168 - Importation of Chinese Sand Pears From China

    Science.gov (United States)

    2011-12-16

    ...; ] DEPARTMENT OF AGRICULTURE Animal and Plant Health Inspection Service 7 CFR Part 319 RIN 0579-AD42 Importation... fungicides. Paragraph (b)(3) would state that, when any sand pears destined for export to the United...

  15. 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.

  16. Simultaneous detection of four viruses affecting apple and pear by molecular hybridization using a polyprobe

    Directory of Open Access Journals (Sweden)

    Thor Vinícius Martins Fajardo

    2014-10-01

    Full Text Available The viruses Apple stem grooving virus (ASGV, Apple chlorotic leaf spot virus (ACLSV, Apple stem pitting virus (ASPV and Apple mosaic virus (ApMV are common in apples and pears and main targets of detection in propagation materials. This study aimed at demonstrating the usefulness of the hybridization method with a non-radioactive probe for simultaneous detection of these four viruses. The sensitivity of this method was sufficiently high enabling the detection of ASGV, ACLSV, ASPV and ApMV in total RNA extracted from infected samples. The probe specificity was confirmed by reaction with homologous viral cDNA, individually cloned for each virus.

  17. Studies on vibration characteristics of a pear using finite element method

    Institute of Scientific and Technical Information of China (English)

    SONG Hui-zhi; WANG Jun; LI Yong-hui

    2006-01-01

    The variation of the vibration characteristics of a Huanghua pear was investigated using finite element simulations. A new image processing technique was used to obtain the unsymmetrical and un-spherical geometrical model of a pear. The vibration characteristics of this type of pear with the correlation of its behavior with geometrical configurations and material characteristics were investigated using numerical modal analysis. The results showed that the eigenfrequency increased with the increasing pear Young's modulus, while decreased with increasing pear density, and decreased with increasing pear volume. The results of this study provided foundation for further investigations of the physical characteristics of fruits and vegetables by using finite element simulations.

  18. Preparing a re-sequencing DNA library of 2 cancer candidate genes using the ligation-by-amplification protocol by two PCR reactions

    Institute of Scientific and Technical Information of China (English)

    SU YeYang; LIN Lin; TIAN Geng; CHEN Chen; LIU Tao; XU Xingya; QI XinPeng; ZHANG XiuQing; YANG HuanMing

    2009-01-01

    To meet the needs of large-scale genomic/genetic studies, the next-generation massively parallelized sequencing technologies provide high throughput, low cost and low labor-intensive sequencing ser-vice, with subsequent bioinformatic software and laboratory methods developed to expand their ap-plications in various types of research. PCR-based genomic/genetic studies, which have significant usage in association studies like cancer research, haven't benefited much from those next-generation sequencing technologies, because the shortgun re-sequencing strategy used by such sequencing machines as the Illumina/Solexa Genome Analyzer may not be applied to direct re-sequencing of short-length target regions like those in PCR-based genomic/genetic studies. Although several meth-ods have been proposed to solve this problem, including microarray-based genomic selections and selector-based technologies, they require advanced equipment and procedures which limit their ap-plications in many laboratories. By contrast, we overcame such potential drawbacks by utilizing a liga-tion by amplification (LBA) protocol, a method using a pair of Universal Adapters to randomly ligate target regions in a two-step-PCR procedure, whose Long LBA products were easily fragmented and sequenced on the next-generation sequencing machine. In this concept-proven study, we chose the consensus coding sequences of two human cancer genes: BRCA1 and BRCA2 as target regions, spe-cifically designed LBA primer pairs to amplify and randomly ligate them. 70 target sequences were successfully amplified and ligated into Long LBA products, which were then fragmented to construct DNA libraries for sequencing on both a conventional Sanger sequencer ABI 3730xl DNA Analyzer and the next-generation 'synthesis by sequencing technology' IlluminalSolexa Genome Analyzer. Bioin-formatic analysis demonstrated the utility and efficiency (including the coverage and depth of each target sequence and the SNPs detection

  19. Preparing a re-sequencing DNA library of 2 cancer candidate genes using the ligation-by-amplification protocol by two PCR reactions

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    To meet the needs of large-scale genomic/genetic studies, the next-generation massively parallelized sequencing technologies provide high throughput, low cost and low labor-intensive sequencing service, with subsequent bioinformatic software and laboratory methods developed to expand their applications in various types of research. PCR-based genomic/genetic studies, which have significant usage in association studies like cancer research, haven’t benefited much from those next-generation sequencing technologies, because the shortgun re-sequencing strategy used by such sequencing machines as the Illumina/Solexa Genome Analyzer may not be applied to direct re-sequencing of short-length target regions like those in PCR-based genomic/genetic studies. Although several methods have been proposed to solve this problem, including microarray-based genomic selections and selector-based technologies, they require advanced equipment and procedures which limit their applications in many laboratories. By contrast, we overcame such potential drawbacks by utilizing a ligation by amplification (LBA) protocol, a method using a pair of Universal Adapters to randomly ligate target regions in a two-step-PCR procedure, whose Long LBA products were easily fragmented and sequenced on the next-generation sequencing machine. In this concept-proven study, we chose the consensus coding sequences of two human cancer genes: BRCA1 and BRCA2 as target regions, specifically designed LBA primer pairs to amplify and randomly ligate them. 70 target sequences were successfully amplified and ligated into Long LBA products, which were then fragmented to construct DNA libraries for sequencing on both a conventional Sanger sequencer ABI 3730xl DNA Analyzer and the next-generation ’synthesis by sequencing technology’ Illumina/Solexa Genome Analyzer. Bioinformatic analysis demonstrated the utility and efficiency (including the coverage and depth of each target sequence and the SNPs detection

  20. Assessing HER2 amplification by IHC, FISH, and real-time polymerase chain reaction analysis (real-time PCR) following LCM in formalin-fixed paraffin embedded tissue from 40 women with ovarian cancer.

    Science.gov (United States)

    Hillig, Thore; Thode, Jørgen; Breinholt, Marie F; Franzmann, Maria-Benedicte; Pedersen, Carsten; Lund, Flemming; Mygind, Henrik; Sölétormos, György; Rudnicki, Martin

    2012-12-01

    We compare HER2 receptor amplification analysis by immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), and real-time polymerase chain reaction (real-time PCR) DNA copy-number assay following laser capture microdissection (LCM) in formalin-fixed paraffin embedded tissue from 40 women with verified ovarian cancer. We speculate that LCM should result in a more accurate assessment of HER2 amplification in our real-time PCR assay compared with IHC and FISH. HER2 overexpression measured by IHC, FISH, or real-time PCR was found in 5.0%, 5.0%, and 22.5%, respectively. HER2 negative results measured by IHC, FISH, or real-time PCR were found in 95%, 92.5%, and 60.0%, respectively. Analysis failed for IHC, FISH, or real-time PCR in 0%, 2.5%, or 17.5% of cases. Concordance between IHC and FISH, IHC and real-time PCR, or FISH and real-time PCR were 89.7%, 72.7%, or 78.1%, respectively. Only few ovarian cancer patients were HER2 overexpressed measured by IHC or FISH and thus could be eligible for antibody-based therapy with trastuzumab (Herceptin). Interestingly, we find an increased number of HER2 positive patients by real-time PCR analysis on microdissected cancer cells, suggesting a number of HER2 positive patients not detected by current methods. Thus, the concept of quantitative measurement of HER2 on microdissected cancer cells should be explored further.

  1. Selective amplification of cDNA sequence from total RNA by cassette-ligation mediated polymerase chain reaction (PCR): application to sequencing 6.5 kb genome segment of hantavirus strain B-1.

    Science.gov (United States)

    Isegawa, Y; Sheng, J; Sokawa, Y; Yamanishi, K; Nakagomi, O; Ueda, S

    1992-12-01

    A method, referred to as cassette-ligation mediated polymerase chain reaction (PCR), has been developed to permit selective and specific amplification of cDNA sequence from total cellular RNA. This technique comprises (i) digestion of cDNA with multiple restriction enzymes, (ii) ligation of cleavage products to double-stranded DNA cassettes possessing a corresponding restriction site and (iii) amplification of cassette-ligated restriction fragments containing a short, known sequence (but not all the other ligation products) by PCR using the specific and cassette primers; the specific primer is designed to prime synthesis from the known sequence of the cDNA whereas the cassette primer anneals to one strand of the cassette. Sequencing from the cassette primer provides information to design a new primer for the next walking step. The amplified cDNA fragments are often larger than the maximum DNA fragments (500-600 bp) that can be sequenced without the need of synthesizing internal sequencing primer. Each of such large cDNA fragments is dissected into smaller DNA fragments by repeating cassette-ligation mediated PCR exploiting different restriction sites and different sets of cassette primers. This dissection process reduces the number of specific primers to a minimum, thereby increasing the speed of sequencing and minimizing the overall cost. We have successfully applied this cDNA walking and sequencing by the cassette-ligation mediated PCR to the sequencing of an entire 6.5 kb genome segment of hantavirus strain B-1.(ABSTRACT TRUNCATED AT 250 WORDS)

  2. Loop-mediated isothermal amplification for detection of nucleic acids.

    Science.gov (United States)

    Tanner, Nathan A; Evans, Thomas C

    2014-01-06

    Sequence-specific isothermal nucleic acid amplification techniques are ideally suited for use in molecular diagnostic applications because they do not require thermal cycling equipment and the reactions are typically fast. One of the most widely cited isothermal techniques is termed loop-mediated isothermal amplification (LAMP). This protocol allows amplification times as fast as 5 to 10 min. Furthermore, various methodologies to detect amplification have been applied to LAMP to increase its utility for the point-of-care market. Basic LAMP protocols are provided herein for detection of specific DNA and RNA targets, along with a method to perform multiplex LAMP reactions, permitting even greater flexibility from this powerful technique.

  3. 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......, while Hum-TH01, HumCD4, and D12S391 were virtually unaffected by low-stringency conditions. Replacement of the Taq DNA polymerase with DNA polymerases with lower processivity resulted in higher levels of extra peaks. Our results support the hypothesis that extra peaks are produced due to slipped...

  4. Bioanalytical applications of isothermal nucleic acid amplification techniques.

    Science.gov (United States)

    Deng, Huimin; Gao, Zhiqiang

    2015-01-01

    The most popular in vitro nucleic acid amplification techniques like polymerase chain reaction (PCR) including real-time PCR are costly and require thermocycling, rendering them unsuitable for uses at point-of-care. Highly efficient in vitro nucleic acid amplification techniques using simple, portable and low-cost instruments are crucial in disease diagnosis, mutation detection and biodefense. Toward this goal, isothermal amplification techniques that represent a group of attractive in vitro nucleic acid amplification techniques for bioanalysis have been developed. Unlike PCR where polymerases are easily deactivated by thermally labile constituents in a sample, some of the isothermal nucleic acid amplification techniques, such as helicase-dependent amplification and nucleic acid sequence-based amplification, enable the detection of bioanalytes with much simplified protocols and with minimal sample preparations since the entire amplification processes are performed isothermally. This review focuses on the isothermal nucleic acid amplification techniques and their applications in bioanalytical chemistry. Starting off from their amplification mechanisms and significant properties, the adoption of isothermal amplification techniques in bioanalytical chemistry and their future perspectives are discussed. Representative examples illustrating the performance and advantages of each isothermal amplification technique are discussed along with some discussion on the advantages and disadvantages of each technique.

  5. Risk assessment and ranking of pesticide residues in Chinese pears

    Institute of Scientific and Technical Information of China (English)

    LI Zhi-xia; LIU Chuan-de; ZHAO Xu-bo; GUO Yong-ze; NIE Ji-yun; YAN Zhen; XU Guo-feng; LI Hai-fei; KUANG Li-xue; PAN Li-gang; XIE Han-zhong; WANG Cheng

    2015-01-01

    The presence of pesticide residues in pears is a serious health concern. This study presents the results from a 2-year investigation (2013–2014) that used gas chromatography, GS/MS and UPLC/MS-MS to measure the levels of 104 pesti-cides in 310 pear samples. In 93.2% of the samples, 43 pesticides were detected, of which the maximum residue levels (MRLs) were exceeded in 2.6% of the samples. Multiple residues (two to eight compounds) were present in 69.7% of the samples; one sample contained nine pesticides and one sample contained 10. Only 6.8% of the samples did not contain residues. To assess the health risks, the pesticide residue data have been combined with daily pear consumption data for children and adult populations. A deterministic model was used to assess the chronic and acute exposures based on the Joint Meeting on Pesticide Residues (JMPR) method. A potential acute risk was demonstrated for children in the case of bifenthrin, which was found to be present at 105.36% of the acute reference dose (ARfD) value. The long-term exposure of the Chinese consumer to pesticide residues through the consumption of raw pears was far below the acceptable daily intake (ADI) criterion. Additionally, the matrix ranking scheme was used to classify risk subgroups of pesticides and pear samples. In general, 95.5% of samples were deemed to be safe and nine pesticides were classiifed as being of a relatively high risk. The ifndings indicated that the occurrence of pesticide residues in pears should not be considered a serious public health problem. Nevertheless, a more detailed study is required for vulnerable consumer groups, especially children. Continuous monitoring of pesticides in pears and tighter regulation of pesticide residue standards are recommended.

  6. Advances in isothermal amplification: novel strategies inspired by biological processes.

    Science.gov (United States)

    Li, Jia; Macdonald, Joanne

    2015-02-15

    Nucleic acid amplification is an essential process in biological systems. The in vitro adoption of this process has resulted in powerful techniques that underpin modern molecular biology. The most common tool is polymerase chain reaction (PCR). However, the requirement for a thermal cycler has somewhat limited applications of this classic nucleic acid amplification technique. Isothermal amplification, on the other hand, obviates the use of a thermal cycler because reactions occur at a single temperature. Isothermal amplification methods are diverse, but all have been developed from an understanding of natural nucleic acid amplification processes. Here we review current isothermal amplification methods as classified by their enzymatic mechanisms. We compare their advantages, disadvantages, efficiencies, and applications. Finally, we mention some new developments associated with this technology, and consider future possibilities in molecular engineering and recombinant technologies that may develop from an appreciation of the molecular biology of natural systems.

  7. Quince 'CPP': new dwarf rootstock for pear trees on organic and high density planting

    OpenAIRE

    Renato Vasconcelos Botelho; Everton Schneider; Danielle Machado; Rafael Piva; Andricia Verlindo

    2012-01-01

    In Brazil, pear production presents the same incipient situation over the last 15 years, due mostly to low production technology. In this context, this study aimed to evaluate the development, growth and production of the pear tree cultivars Cascatense, Tenra and Hosui grafted on 'CPP' quince rootstock, using 'FT' pear as interstem. This trial was carried out in Guarapuava, State of Paraná, Southern region of Brazil, by five productive cycles. The pear trees were planted in September of 2004,...

  8. 7 CFR 319.56-29 - Ya variety pears from China.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 5 2010-01-01 2010-01-01 false Ya variety pears from China. 319.56-29 Section 319.56... variety pears from China. Ya variety pears may be imported into the United States from China only in... organization (NPPO) of China in an APHIS-approved export growing area in the Hebei or Shandong Provinces....

  9. 7 CFR 917.21 - Nomination of Pear Commodity Committee members.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false Nomination of Pear Commodity Committee members. 917.21... AND PEACHES GROWN IN CALIFORNIA Order Regulating Handling Administrative Bodies § 917.21 Nomination of Pear Commodity Committee members. Nominations for membership on the Pear Commodity Committee shall...

  10. Rapid and efficient identification of the mouse leptin receptor mutation (C57BL/KsJ-db/db) by tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) analysis.

    Science.gov (United States)

    Jung, Harry; Nam, Hajin; Suh, Jun-Gyo

    2016-03-01

    The C57BLKS/J-Lepr(db) mouse has a point mutation in the leptin receptor gene and is one of the most useful animal model for non-insulin dependent diabetes mellitus in human. Since the homozygote of C57BLKS/J-Lepr(db) mouse is infertile, detection of point mutation in the leptin receptor gene is important for efficient maintaining strains as well as mass production of homozygotes. To develop a rapid and efficient genotyping method for C57BLKS/J-Lepr(db) mouse, the tetra-primer amplification-refractory mutation system polymerase chain reaction (ARMS-PCR) was used. The 407 and 199 bp PCR products were amplified from normal (+/+) mice; while the 407 and 268 bp PCR products were amplified from homozygotes (db/db) mice; and the 407, 268, and 199 bp PCR products were amplified from heterozygotes (db/+) mice. This result showed that the tetra-primer ARMS-PCR analysis by us is suitable to detect point mutation of the leptin receptor gene. Taken together, our method will dramatically reduce animal use for maintenance of strains as well as production of homozygote in the C57BLKS/J-Lepr(db) strains.

  11. Practical Prediction of Ten Common Streptococcus pneumoniae Serotypes/Serogroups in One PCR Reaction by Multiplex Ligation-Dependent Probe Amplification and Melting Curve (MLPA-MC Assay in Shenzhen, China.

    Directory of Open Access Journals (Sweden)

    Lijuan Wu

    Full Text Available Streptococcus pneumoniae has more than 95 distinct serotypes described to date. However, only certain serotypes are more likely to cause pneumococcal diseases. Thus serotype surveillance is important for vaccine formula design as well as in post-vaccine serotype shift monitor. The goal of this study was to develop a practical screening assay for ten Shenzhen China common pneumococcal serotypes/serogroups in one molecular reaction.A molecular assay, based on multiplex ligation-dependent probe amplification (MLPA and melting curve (MC analysis, was developed in an integrated approach (MLPA-MC for the detection of ten capsular serotypes/serogroups 4, 6 (6A/6B/6C/6D, 9V/9A, 14, 15F/15A, 15B/15C, 18 (18F/18A/18B/18C, 19F, 19A and 23F. We designed serotype/serogroup-specific MLPA probes and fluorescent detection probes to discriminate the different serotypes/serogroups in one molecular reaction. The three steps of MLPA-MC assay are continuous reactions in one well detected by LightCycler 480. A total of 210 S. pneumoniae isolates from our local Maternity and Child Health Hospital were randomly chosen to evaluate the assay against published multiplex PCR assays.Our results showed that 198 (94.3% of S. pneumoniae isolates were type-able by our assays and the results were in complete concordance with the published multiplex PCRs. Using the MLPA-MC assay, 96 S. pneumoniae isolates could be typed within 3 hours with limited hands-on time. This serotype/serogroup-screening assay can be easily modified or extended by modification of the serotype/serogroup-specific MLPA probes combinations according to the needs of different laboratories.We recommend use of this assay as a starting point for screening serotype/serogroup frequencies. There is a need for this assay to be combined with other molecular typing assays, like published serotype specific PCRs, or even the Quellung reaction for serotype confirmation.

  12. 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.

  13. Interactive Effects of Growth Regulators, Carbon Sources, pH on Plant Regeneration and Assessment of Genetic Fidelity Using Single Primer Amplification Reaction (SPARS) Techniques in Withania somnifera L.

    Science.gov (United States)

    Fatima, Nigar; Ahmad, Naseem; Ahmad, Iqbal; Anis, Mohammad

    2015-09-01

    An improved and methodical in vitro shoot morphogenic approach through axillary bud multiplication was established in a drug yielding plant, Withania somnifera L. Effects of plant growth regulators [6-benzyladenine (BA), kinetin (Kin), 2-isopentenyladenine (2iP), and thidiazuron (TDZ)] either singly or in combination with α-napthalene acetic acid (NAA), indole-3-butyric acid (IBA), and indole-3-acetic acid (IAA) in Murashige and Skoog (MS) medium were tested. The highest regeneration frequency (90 %) with optimum number of shoots (32 ± 0.00)/explant were obtained on MS medium fortified with 2.5 μM 6-benzyladenine (BA) and 0.5 μM NAA and 30 g/l sucrose at pH 5.8. Among the tried TDZ concentrations, 0.5 μM resulted in maximum number of shoots (20.4 ± 0.40)/explant after 4 weeks of exposure. The proliferating shoot cultures established by repeated subculturing of the mother explants on the hormone-free medium produced the highest shoot number (29.4 ± 0.40) with shoot length (6.80 ± 0.12 cm)/explant at fourth subculture passage, which a decline in shoot proliferation was recorded. Different concentrations of NAA were tested for ex vitro rooting of microshoots. The maximum percentage of rooting 100 % with maximum roots (18.3 ± 0.1) was achieved in soilrite when basal portion of the microshoots were treated with 200 μM (NAA) for 15 min per shoot. The plantlets went through hardening phase in a growth chamber, prior to ex vitro transfer. The PCR-based single primer amplification reaction (SPAR) methods which include random amplified polymorphic DNA (RAPD) and direct amplification of minisatellite DNA (DAMD) markers has been used for assessment of genetic stability of micropropagated plantlets. No variation was observed in DNA fingerprinting patterns among the micropropagated and the donor plants illustrating their genetic uniformity.

  14. The Seneca Amplification Construction

    Directory of Open Access Journals (Sweden)

    Wallace Chafe

    2012-01-01

    Full Text Available The polysynthetic morphology of the Northern Iroquoian languages presents a challenge to studies of clause combining. The discussion here focuses on a Seneca construction that may appear within a single clause but may also straddle clause boundaries. It amplifies the information provided by a referent, here called the trigger, that is introduced by the pronominal prefix within a verb or occasionally in some other way. The particle neh signals that further information about that referent will follow. This construction is found at four levels of syntactic complexity. At the first level the trigger and its amplification occur within the same prosodic phrase and the amplification is a noun. At the second level the amplification occurs in a separate prosodic phrase but remains a noun. At the third level the amplification exhibits verb morphology but has been lexicalized with a nominal function. At the fourth level the amplification functions as a full clause and neh serves as a marker of clause combining. Several varieties of amplification are discussed, as are cases in which the speaker judges that no amplification is needed. It is suggested that the typologically similar Caddo language illustrates a situation in which this construction could never arise, simply because Caddo verbs lack the pronominal element that triggers the construction in Seneca.

  15. Sequence Characterization and Spatiotemporal Expression Patterns of PbS26-RNase Gene in Chinese White Pear (Pyrus bretschneideri

    Directory of Open Access Journals (Sweden)

    Lin Zhang

    2014-01-01

    Full Text Available Many flowering plants exhibit an important intraspecific reproductive barrier phenomenon, that is, self-incompatibility (SI, in which S-RNase genes play a significant role. To clarify the specific function of S-RNase genes in Chinese pears, the full length cDNA of PbS26-RNase was isolated by rapid amplification of cDNA ends (RACE technology from Chinese white pear (Pyrus bretschneideri cultivar “Hongpisu.” The cDNA sequence for PbS26-RNase was deposited in GenBank under accession number EU081888. At the amino acid level, the PbS26-RNase displayed the highest similarity (96.9% with PcSa-RNase of P. communis, and only seven amino acid differences were present in the two S-RNases. Phylogenetic analysis of rosaceous S-RNases indicated that the PbS26-RNase clustered with maloideous S-RNases, forming a subfamily-specific not a species-specific group. The PbS26-RNase gene was specifically expressed in the style but not other tissues/organs. The expression level of the PbS26-RNase gene rapidly increased at bell balloon stage (BBS, and then it dropped after pollination. However, the abundance of the PbS26-RNase gene transcript in the style was greater after cross-pollination than after self-pollination. In addition, a method for rapidly detecting the PbS26-RNase gene was developed via allele-specific primers design. The present study could provide a scientific basis for fully clarifying the mechanism of pear SI at the molecular level.

  16. Effect of Extrusion Cooking on Bioactive Compounds in Encapsulated Red Cactus Pear Powder

    Directory of Open Access Journals (Sweden)

    Martha G. Ruiz-Gutiérrez

    2015-05-01

    Full Text Available Red cactus pear has significant antioxidant activity and potential as a colorant in food, due to the presence of betalains. However, the betalains are highly thermolabile, and their application in thermal process, as extrusion cooking, should be evaluated. The aim of this study was to evaluate the effect of extrusion conditions on the chemical components of red cactus pear encapsulated powder. Cornstarch and encapsulated powder (2.5% w/w were mixed and processed by extrusion at different barrel temperatures (80, 100, 120, 140 °C and screw speeds (225, 275, 325 rpm using a twin-screw extruder. Mean residence time (trm, color (L*, a*, b*, antioxidant activity, total polyphenol, betacyanin, and betaxanthin contents were determined on extrudates, and pigment degradation reaction rate constants (k and activation energies (Ea were calculated. Increases in barrel temperature and screw speed decreased the trm, and this was associated with better retentions of antioxidant activity, total polyphenol, betalain contents. The betacyanins k values ranged the −0.0188 to −0.0206/s and for betaxanthins ranged of −0.0122 to −0.0167/s, while Ea values were 1.5888 to 6.1815 kJ/mol, respectively. The bioactive compounds retention suggests that encapsulated powder can be used as pigments and to provide antioxidant properties to extruded products.

  17. Effect of extrusion cooking on bioactive compounds in encapsulated red cactus pear powder.

    Science.gov (United States)

    Ruiz-Gutiérrez, Martha G; Amaya-Guerra, Carlos A; Quintero-Ramos, Armando; Pérez-Carrillo, Esther; Ruiz-Anchondo, Teresita de J; Báez-González, Juan G; Meléndez-Pizarro, Carmen O

    2015-05-18

    Red cactus pear has significant antioxidant activity and potential as a colorant in food, due to the presence of betalains. However, the betalains are highly thermolabile, and their application in thermal process, as extrusion cooking, should be evaluated. The aim of this study was to evaluate the effect of extrusion conditions on the chemical components of red cactus pear encapsulated powder. Cornstarch and encapsulated powder (2.5% w/w) were mixed and processed by extrusion at different barrel temperatures (80, 100, 120, 140 °C) and screw speeds (225, 275, 325 rpm) using a twin-screw extruder. Mean residence time (trm), color (L*, a*, b*), antioxidant activity, total polyphenol, betacyanin, and betaxanthin contents were determined on extrudates, and pigment degradation reaction rate constants (k) and activation energies (Ea) were calculated. Increases in barrel temperature and screw speed decreased the trm, and this was associated with better retentions of antioxidant activity, total polyphenol, betalain contents. The betacyanins k values ranged the -0.0188 to -0.0206/s and for betaxanthins ranged of -0.0122 to -0.0167/s, while Ea values were 1.5888 to 6.1815 kJ/mol, respectively. The bioactive compounds retention suggests that encapsulated powder can be used as pigments and to provide antioxidant properties to extruded products.

  18. Amplification of NOON States

    CERN Document Server

    Agarwal, G S; Rai, Amit

    2009-01-01

    We examine the behavior of a Non Gaussian state like NOON state under phase insensitive amplification. We derive analytical result for the density matrix of the NOON state for arbitrary gain of the amplifier. We consider cases of both symmetric and antisymmetric amplification of the two modes of the NOON state. We quantitatively evaluate the loss of entanglement by the amplifier in terms of the logarithmic negativity parameter. We find that NOON states are more robust than their Gaussian counterparts.

  19. Amplification of NOON States

    OpenAIRE

    2009-01-01

    We examine the behavior of a Non Gaussian state like NOON state under phase insensitive amplification. We derive analytical result for the density matrix of the NOON state for arbitrary gain of the amplifier. We consider cases of both symmetric and antisymmetric amplification of the two modes of the NOON state. We quantitatively evaluate the loss of entanglement by the amplifier in terms of the logarithmic negativity parameter. We find that NOON states are more robust than their Gaussian coun...

  20. Accuracy of marker analysis, quantitative real-time polymerase chain reaction, and multiple ligation-dependent probe amplification to determine SMN2 copy number in patients with spinal muscular atrophy.

    Science.gov (United States)

    Alías, Laura; Bernal, Sara; Barceló, Maria J; Also-Rallo, Eva; Martínez-Hernández, Rebeca; Rodríguez-Alvarez, Francisco J; Hernández-Chico, Concepción; Baiget, Montserrat; Tizzano, Eduardo F

    2011-09-01

    Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder caused by absence of or mutations in the survival motor neuron1 gene (SMN1). All SMA patients have a highly homologous copy of SMN1, the SMN2 gene. Severe (type I) SMA patients present one or two SMN2 copies, whereas milder chronic forms (type II-III) usually have three or four SMN2 copies. SMN2 dosage is important to stratify patients for motor function tests and clinical trials. Our aim was to compare three methods, marker analysis, real-time quantitative polymerase chain reaction using the LightCycler instrument, and multiple ligation-dependent probe amplification (MLPA), to characterize their accuracy in quantifying SMN2 genes. We studied a group of 62 genetically confirmed SMA patients, 54 with homozygous absence of exons 7 and 8 of SMN1 and 8 with SMN2-SMN1 hybrid genes. A complete correlation using the three methods was observed in 32 patients (51.6%). In the remaining 30 patients, discordances between the three methods were found, including under or overestimation of SMN2 copies by marker analysis with respect to the quantitative methods (LightCycler and MLPA) because of lack of informativeness of markers, 3' deletions of SMN genes, and breakpoints in SMN2-SMN1 hybrid genes. The technical limitations and advantages and disadvantages of these methods are discussed. We conclude that the three methods complement each other in estimating the SMN2 copy number in most cases. However, MLPA offers additional information to characterize SMA cases with particular rearrangements such as partial deletions and hybrid genes.

  1. Identification of deletions and duplications in the Duchenne muscular dystrophy gene and female carrier status in western India using combined methods of multiplex polymerase chain reaction and multiplex ligation-dependent probe amplification

    Directory of Open Access Journals (Sweden)

    Rashna S Dastur

    2011-01-01

    Full Text Available Background: The technique of multiplex ligation-dependent probe amplification (MLPA assay is an advanced technique to identify deletions and duplications of all the 79 exons of DMD gene in patients with Duchenne/Becker muscular dystrophy (DMD/BMD and female carriers. Aim: To use MLPA assay to detect deletions which remained unidentified on multiplex polymerase chain reaction (mPCR analysis, scanning 32 exons of the "hot spot" region. Besides knowing the deletions and/or duplications, MLPA was also used to determine the carrier status of the females at risk. Materials and Methods: Twenty male patients showing no deletions on mPCR and 10 suspected carrier females were studied by MLPA assay using P-034 and P-035, probe sets (MRC Holland covering all the 79 exons followed by capillary electrophoresis on sequencing system. Results: On MLPA analysis, nine patients showed deletions of exons other than 32 exons screened by mPCR represented by absence of peak. Value of peak areas were double or more in four patients indicating duplications of exons. Carrier status was confirmed in 50% of females at risk. Conclusion: Combining the two techniques, mPCR followed by MLPA assay, has enabled more accurate detection and extent of deletions and duplications which otherwise would have remained unidentified, thereby increasing the mutation pick up rate. These findings have also allowed prediction of expected phenotype. Determining carrier status has a considerable significance in estimating the risk in future pregnancies and prenatal testing options to limit the birth of affected individuals.

  2. 7 CFR 927.125 - Fresh pear reports.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false Fresh pear reports. 927.125 Section 927.125 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing... destination by city and state or city and country; (5) The name and address of such handler; and (6)...

  3. Authenticity analysis of pear juice employing chromatographic fingerprinting.

    Science.gov (United States)

    Willems, Jamie L; Low, Nicholas H

    2014-12-01

    Pear juice is predominately composed of carbohydrates/polyols (>95% of the total soluble solids), making it susceptible to adulteration by the addition of less expensive commercial sweeteners. In this research, the major carbohydrate and polyol (fructose, glucose, sucrose, and sorbitol) content of 32 pure pear juices representing five world producing regions and three years of production was determined. Additionally, methods employing oligosaccharide profiling to detect the debasing of these samples with four commercial sweeteners (HFCS 55 and 90, TIS, and HIS) were developed using capillary gas chromatography with flame ionization detection (CGC-FID) and high-performance liquid chromatography with pulsed amperometric detection (HPAE-PAD). Detection limits for the four commercial sweeteners ranged from 0.5 to 5.0% (v/v). In addition, the developed CGC-FID method could be used to (a) detect the addition of pear to apple juice via arbutin detection and (b) determine if a pear juice was produced using enzymatic liquefaction via the presence of O-β-d-glucopyranosyl-(1→4)-d-glucopyranose (cellobiose), all within a single chromatographic analysis.

  4. 21 CFR 145.176 - Artificially sweetened canned pears.

    Science.gov (United States)

    2010-04-01

    ... thickened with pectin and may contain any mixture of any edible organic salt or salts and any edible organic..., as prescribed for canned pears by § 145.175(a). If the packing medium is thickened with pectin, the label shall bear the statement “thickened with pectin”. When any organic salt or acid or any mixture...

  5. Mineral nutrition influences physiological responses of pear in vitro

    Science.gov (United States)

    Physiological disorders such as callus, shoot tip necrosis and hyperhydricity are some of the most difficult challenges in micropropagation and their causes are not well understood. A comprehensive medium optimization study to improve the growth of pear shoot cultures was also designed to determine ...

  6. 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.

  7. 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.

  8. Research of Nanguo Pear Polyphenol Oxidase%南国梨多酚氧化酶的研究

    Institute of Scientific and Technical Information of China (English)

    何晓蒙; 孔繁东

    2016-01-01

    Fresh Nanguo pear as raw material, catechol as substrate, using spectrophotometric study of south-ern pear enzymatic characteristics of polyphenol oxidase. The results showed that:pH and temperature had a significant effect on fresh Nanguo pear PPO activity and the optimum pH was 6.8, fresh pear PPO optimum temperature was 40℃. Further study of the inhibitor of ascorbic acid, citric acid, sodium bisulfite, L-cys-teine, EDTA-2Na and various metal salts [CuSO4, CaCl2, Na2SO4, Al(NO3)3, CdCl2, HgCl2, MnCl2] effect of southern pear PPO, where 0.1 mmol/L of ascorbic acid on fresh pear PPO activity was significantly inhibited by up to 50 %. Nanguo pear PPO enzymatic catalytic substrate catechol reaction kinetics and the Michaelis-Menten equation height accord, R2=0.998, its kinetic equation 1/[V]=0.002/[S]+0.019, maximum reaction rate Vmax to 52.632 U/min,the Michaelis constant Km of 0.010 5 mol/L.%以新鲜南国梨果实为原料,邻苯二酚为底物,采用分光光度法研究南国梨多酚氧化酶的酶学特性。结果表明:pH和温度对新鲜南国梨PPO活性有明显的影响,最适pH为6.8,新鲜南国梨PPO最适温度为40℃。进一步研究了抑制剂抗坏血酸、柠檬酸、亚硫酸氢钠、L-半胱氨酸、EDTA-2Na和各种金属盐[CuSO4、CaCl2、Na2SO4、Al(NO3)3、CdCl2、HgCl2、MnCl2]对南国梨PPO的影响,其中0.1 mmol/L的抗坏血酸对新鲜南国梨PPO活性有明显抑制作用高达50%。南国梨PPO催化底物邻苯二酚的酶促反应动力学与米氏方程高度符合,R2=0.998,其动力学方程为1/[V]=0.002/[S]+0.019,最大反应速率Vmax为52.632 U/min,米氏常数Km为0.0105 mol/L。

  9. Quality control for quantitative PCR based on amplification compatibility test.

    Science.gov (United States)

    Tichopad, Ales; Bar, Tzachi; Pecen, Ladislav; Kitchen, Robert R; Kubista, Mikael; Pfaffl, Michael W

    2010-04-01

    Quantitative qPCR is a routinely used method for the accurate quantification of nucleic acids. Yet it may generate erroneous results if the amplification process is obscured by inhibition or generation of aberrant side-products such as primer dimers. Several methods have been established to control for pre-processing performance that rely on the introduction of a co-amplified reference sequence, however there is currently no method to allow for reliable control of the amplification process without directly modifying the sample mix. Herein we present a statistical approach based on multivariate analysis of the amplification response data generated in real-time. The amplification trajectory in its most resolved and dynamic phase is fitted with a suitable model. Two parameters of this model, related to amplification efficiency, are then used for calculation of the Z-score statistics. Each studied sample is compared to a predefined reference set of reactions, typically calibration reactions. A probabilistic decision for each individual Z-score is then used to identify the majority of inhibited reactions in our experiments. We compare this approach to univariate methods using only the sample specific amplification efficiency as reporter of the compatibility. We demonstrate improved identification performance using the multivariate approach compared to the univariate approach. Finally we stress that the performance of the amplification compatibility test as a quality control procedure depends on the quality of the reference set.

  10. 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.

  11. PEAR AS A SOURCE OF BIOLOGICALLY ACTIVE SUBSTANCES FOR PRODUCTS OF FUNCTIONAL PURPOSES

    Directory of Open Access Journals (Sweden)

    Rodionova L. Y.

    2015-01-01

    Full Text Available Biochemical quantitative and qualitative indices of pear fruit have been investigated in six varieties of pears grown in Prikybanskoy horticultural zone of the Krasnodar region. The investigation has been done with pear fruit in the stage of maturity for harvesting and after 90 days after storage in refrigerator. Quantitative content of dry matter, sugars, vitamins C and P and fraction pectin content in fruits and squeezing of fruits as well as changes in the process of storage have been established

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

    OpenAIRE

    2015-01-01

    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 physicochemica...

  13. 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

  14. Economical effectiveness of vegetative pear nurseries in Albania

    Directory of Open Access Journals (Sweden)

    Bardhosh Ferraj

    2013-02-01

    Full Text Available Sapling production on vegetative rootsctock is considered as an important agronomic activity while Albanian arboriculture is being oriented towards the world contemporary development. The paper presents the evaluation of economical effectiveness of the vegetative pear nurseries, since the evaluation of the increase of economical effectiveness and farm productivity as a real potential of Albanian farmers. The experiment was carried out during two consecutive years, 2009-2010, by the Department of Horticulture at Agricultural University of Tirana in collaboration with a certified national nursery. A randomized complete block design (RCBD with 4 replications and 6 variants with a plot size of 50 saplings for variant in each replication was used. Pear cultivars Abate Fetel, Williams and Koshia used as scions grafted over seedy rootsctock of wild pear and vegetative rootsctock of quince clone Anger, (EM – A, were compared. The data showed that different rootstocks affected sapling features and quality. The use of quince vegetative rootstock EM-A provided the highest values of grafting catching rate of 93.7% (V2,V4,V6 and 95.3% standard saplings of both scions (V2,V4,V6. According to the official standards of the Albanian government, considering the qualitative aspect, both pear cultivars grafted over EM-A rootstocks provided higher qualitative saplings. So, for variants V2 and V4, saplings with 2-3 sceletal branches represented 88.6% and 84.7%, respectively; while saplings with main shoot length of 31-40 cm for variants V2, V4, V6 represented 18.1%, 23.5% and 24.3%. The achieved results confirms the need of spreading and widely use of “mother“ plots for vegetative rootstock production, beside the fact that this sapling category is ready to be planted in open fields one year earlier than saplings with seedy rootstock. The two years data were confirmed statistically by LSD and ANOVA tests.

  15. Maatregelen tegen Pear Decline Phytoplasma infectie via enten

    OpenAIRE

    Dees, R.H.L.; Kock, de, M.J.D.

    2014-01-01

    Pear Decline is een ziekte bij perenbomen en wordt veroorzaakt door een fytoplasma. Zieke perenbomen worden gekenmerkt door een vervroegde en veelal intensieve roodverkleuring van de bladeren. Bij een ernstige aantasting sterft de boom af. Bomen gaan echter niet altijd dood. Sommige bomen groeien door de ziekte heen en verliezen het fytoplasma in de loop van de tijd. Het fytoplasma wordt overgedragen door de perenbladvlo (Cacopsylla pyri), maar kan ook tijdens het handmatig vermeerderen van p...

  16. Antioxidant and Anticlastogenic Capacity of Prickly Pear Juice

    Science.gov (United States)

    Madrigal-Santillán, Eduardo; García-Melo, Fernando; Morales-González, José A.; Vázquez-Alvarado, Patricia; Muñoz-Juárez, Sergio; Zuñiga-Pérez, Clara; Sumaya-Martínez, Maria Teresa; Madrigal-Bujaidar, Eduardo; Hernández-Ceruelos, Alejandra

    2013-01-01

    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. PMID:24145870

  17. 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.

  18. Determination of antioxidant constituents in cactus pear fruits.

    Science.gov (United States)

    Fernández-López, José A; Almela, Luís; Obón, José M; Castellar, Rosario

    2010-09-01

    An analytical study was carried out on the presence of antioxidant constituents and the in vitro antioxidant capacity in the extracts of three species of Spanish red-skinned cactus pear fruits (Opuntia ficus-indica, Opuntia undulata and Opuntia stricta). The cactus pear fruit extracts were analyzed for determined constituents: ascorbic acid, flavonoids (quercetin, isorhamnetin, myricetin, kaempferol and luteolin), betalains, taurine, total carotenoids and total phenolics. The antioxidant capacity was assessed by means of two different methods: the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (Trolox equivalent antioxidant capacity) method and the 2,2-diphenyl-1-picrylhydrazyl radical method. Opuntia ficus-indica fruit extract had the strongest antioxidant capacity and taurine content. O. stricta fruits were the richest in ascorbic acid and total phenolics, whereas O. undulata fruits showed the highest carotenoid content. Quercetin and isorhamnetin were the main flavonoids detected. This study provides basic information on the presence of bioactive compounds and antioxidant capacity in extracts of cactus pear fruits, in order to consider these extracts as ingredient for the production of health-promoting food.

  19. miRNA expression during prickly pear cactus fruit development.

    Science.gov (United States)

    Rosas-Cárdenas, Flor de Fátima; Caballero-Pérez, Juan; Gutiérrez-Ramos, Ximena; Marsch-Martínez, Nayelli; Cruz-Hernández, Andrés; de Folter, Stefan

    2015-02-01

    miRNAs are a class of small non-coding RNAs that regulate gene expression. They are involved in the control of many developmental processes, including fruit development. The increasing amount of information on miRNAs, on their expression, abundance, and conservation between various species, provides a new opportunity to study the role of miRNAs in non-model plant species. In this work, we used a combination of Northern blot and tissue print hybridization analysis to identify conserved miRNAs expressed during prickly pear cactus (Opuntia ficus indica) fruit development. Comparative profiling detected the expression of 34 miRNAs, which were clustered in three different groups that were associated with the different phases of fruit development. Variation in the level of miRNA expression was observed. Gradual expression increase of several miRNAs was observed during fruit development, including miR164. miR164 was selected for stem-loop RT-PCR and for a detailed spatial-temporal expression analysis. At early floral stages, miR164 was mainly localized in meristematic tissues, boundaries and fusion zones, while it was more homogenously expressed in fruit tissues. Our results provide the first evidence of miRNA expression in the prickly pear cactus and provide the basis for future research on miRNAs in Opuntia. Moreover, our analyses suggest that miR164 plays different roles during prickly pear cactus fruit development.

  20. A dual amplification fluorescent strategy for sensitive detection of DNA methyltransferase activity based on strand displacement amplification and DNAzyme amplification.

    Science.gov (United States)

    Cui, Wanling; Wang, Lei; Jiang, Wei

    2016-03-15

    DNA methyltransferase (MTase) plays a critical role in many biological processes and has been regarded as a predictive cancer biomarker and a therapeutic target in cancer treatment. Sensitive detection of DNA MTase activity is essential for early cancer diagnosis and therapeutics. Here, we developed a dual amplification fluorescent strategy for sensitive detection of DNA MTase activity based on strand displacement amplification (SDA) and DNAzyme amplification. A trifunctional double-stranded DNA (dsDNA) probe was designed including a methylation site for DNA MTase recognition, a complementary sequence of 8-17 DNAzyme for synthesizing DNAzyme, and a nicking site for nicking enzyme cleavage. Firstly, the trifunctional dsDNA probe was methylated by DNA MTase to form the methylated dsDNA. Subsequently, HpaII restriction endonuclease specifically cleaved the residue of unmethylated dsDNA. Next, under the action of polymerase and nicking enzyme, the methylared dsDNA initiated SDA, releasing numbers of 8-17 DNAzymes. Finally, the released 8-17 DNAzymes triggered DNAzyme amplification reaction to induce a significant fluorescence enhancement. This strategy could detect DNA MTase activity as low as 0.0082U/mL. Additionally, the strategy was successfully applied for evaluating the inhibitions of DNA MTase using two anticancer drugs, 5-azacytidine and 5-aza-2'-deoxycytidine. The results indicate the proposed strategy has a potential application in early cancer diagnosis and therapeutics.

  1. Native pears of Sardinia affect Penicillium expansum pathogenesis.

    Science.gov (United States)

    Cubaiu, L; Azara, E; Ladu, G; Venditti, T; D'Hallewin, G

    2013-01-01

    Penicillium expansum causes blue mould rot, a serious post-harvest disease of pome fruits and is the main producer of the mycotoxin patulin. The occurrence of natural resistance against different hostpathogens, has been evidenced in some pear accessions of the Sardinian germoplasm. The aim of this research was to correlate P. expansum growth and patulin production on these indigenous pear accessions. In vitro and in vivo experiments were carried out with seven accessions ('Sarmentina', 'Vacchesa', 'De Puleu', 'De su Duca', 'Natalina', 'Oliena', 'Laconi 5') belonging to the CNR-ISPA ex situ collection and one national control cultivar ('Abate'). A wild type P. expansum from our collection was isolated from blue mould-decayed Sardinian pear fruit and selected for its aggressiveness and patulin production. The in vivo assay was carried out using 5 x 2 cm (Ø x thickness) sterilized fruit discs wounded and inoculated by a 10(5)UFC/mL concentration of P. expansum. Fruit discs were incubated at 23 degrees C for 7 days before analysis. The in vitro experiments, aimed at monitoring over time P. expansum mycelial growth and patulin accumulation, were performed with a standard medium (PDA) and a pear puree Agar Medium (PAM). Petri dishes with PDA and PAM were inoculated centrally with P. expansum conidia (10(5)UFC/ml) and then incubated at 23 degrees C for 7 days. Mycelial growth on Sardinian PAMs was inhibited in comparison to 'Abate' PAM and PDA. In particular, the accessions 'Sarmentina' and 'Vacchesa' showed the maximum inhibitory activity both in vitro and in vivo. Patulin production was detected by high-pressure liquid chromatography-mass spectrometry. The mycotoxin concentration in Sardinian PAMs was lower than that detected in PDA medium, pointing out a positive correlation between fungal growth inhibition and patulin production. The lowest concentration of patulin was found in 'Sarmentina' PAM. Based on these findings, some of Sardinian pear accessions seems to

  2. Identification of Pyrus single nucleotide polymorphisms (SNPs) and evaluation for genetic mapping in European pear and interspecific Pyrus hybrids.

    Science.gov (United States)

    Montanari, Sara; Saeed, Munazza; Knäbel, Mareike; Kim, YoonKyeong; Troggio, Michela; Malnoy, Mickael; Velasco, Riccardo; Fontana, Paolo; Won, KyungHo; Durel, Charles-Eric; Perchepied, Laure; Schaffer, Robert; Wiedow, Claudia; Bus, Vincent; Brewer, Lester; Gardiner, Susan E; Crowhurst, Ross N; Chagné, David

    2013-01-01

    We have used new generation sequencing (NGS) technologies to identify single nucleotide polymorphism (SNP) markers from three European pear (Pyrus communis L.) cultivars and subsequently developed a subset of 1096 pear SNPs into high throughput markers by combining them with the set of 7692 apple SNPs on the IRSC apple Infinium® II 8K array. We then evaluated this apple and pear Infinium® II 9K SNP array for large-scale genotyping in pear across several species, using both pear and apple SNPs. The segregating populations employed for array validation included a segregating population of European pear ('Old Home'×'Louise Bon Jersey') and four interspecific breeding families derived from Asian (P. pyrifolia Nakai and P. bretschneideri Rehd.) and European pear pedigrees. In total, we mapped 857 polymorphic pear markers to construct the first SNP-based genetic maps for pear, comprising 78% of the total pear SNPs included in the array. In addition, 1031 SNP markers derived from apple (13% of the total apple SNPs included in the array) were polymorphic and were mapped in one or more of the pear populations. These results are the first to demonstrate SNP transferability across the genera Malus and Pyrus. Our construction of high density SNP-based and gene-based genetic maps in pear represents an important step towards the identification of chromosomal regions associated with a range of horticultural characters, such as pest and disease resistance, orchard yield and fruit quality.

  3. Identification of Pyrus single nucleotide polymorphisms (SNPs and evaluation for genetic mapping in European pear and interspecific Pyrus hybrids.

    Directory of Open Access Journals (Sweden)

    Sara Montanari

    Full Text Available We have used new generation sequencing (NGS technologies to identify single nucleotide polymorphism (SNP markers from three European pear (Pyrus communis L. cultivars and subsequently developed a subset of 1096 pear SNPs into high throughput markers by combining them with the set of 7692 apple SNPs on the IRSC apple Infinium® II 8K array. We then evaluated this apple and pear Infinium® II 9K SNP array for large-scale genotyping in pear across several species, using both pear and apple SNPs. The segregating populations employed for array validation included a segregating population of European pear ('Old Home'×'Louise Bon Jersey' and four interspecific breeding families derived from Asian (P. pyrifolia Nakai and P. bretschneideri Rehd. and European pear pedigrees. In total, we mapped 857 polymorphic pear markers to construct the first SNP-based genetic maps for pear, comprising 78% of the total pear SNPs included in the array. In addition, 1031 SNP markers derived from apple (13% of the total apple SNPs included in the array were polymorphic and were mapped in one or more of the pear populations. These results are the first to demonstrate SNP transferability across the genera Malus and Pyrus. Our construction of high density SNP-based and gene-based genetic maps in pear represents an important step towards the identification of chromosomal regions associated with a range of horticultural characters, such as pest and disease resistance, orchard yield and fruit quality.

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

    Science.gov (United States)

    2013-08-28

    ... AGENCY 40 CFR Part 180 Ethyl-2E,4Z-Decadienoate (Pear Ester); Exemption From the Requirement of a...-2E,4Z- decadienoate (pear ester) in or on all food commodities. This regulation eliminates the need to establish a maximum permissible level for residues of ethyl-2E,4Z-decadienoate (pear ester)....

  5. Biological Control of Phacidiopycnis Rot in ‘d’Anjou’ Pears

    Science.gov (United States)

    Phacidiopycnis rot, caused by Phacidiopycnis piri, is a recently reported postharvest fruit rot disease of pears (Pyrus) in the U.S. and a major disease of ‘d’Anjou’ pears grown in Washington State. Phacidiopycnis rot can originate from infection of wounds on the fruit. In this study, two biocontrol...

  6. Alternaria alternata, causal agent of dead (dormant) flower bud disease of pear

    NARCIS (Netherlands)

    Wenneker, M.; Tjou-Tam-Sin, L.T.; Bruggen, van A.S.; Vink, P.

    2006-01-01

    Dead (dormant) flower buds of pear are an important phenomenon in pear production in the Netherlands. Vigourous or unbalanced tree growth and Pseudomonas syringae pv. syringae are mentioned as likely causes of dead flower buds. Several tree growth control treatments including ethephon, Regalis (Proh

  7. 76 FR 27848 - Pears Grown in Oregon and Washington; Amendment To Allow Additional Exemptions

    Science.gov (United States)

    2011-05-13

    ... provided an exemption for consumer- direct sales of up to 220 pounds of fresh pears per transaction, for... the interim rule that added an exemption for consumer-direct sales of up to 220 pounds of fresh pears..., roadside stands, or farmers' markets. These consumer-direct sales are exempt from the marketing...

  8. Combined morphological and molecular approach for identification of Stemphylium vesicarium inoculum in pear orchards.

    Science.gov (United States)

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

    2015-03-01

    Stemphylium vesicarium is the causal agent of brown spot of pear, an important disease reported in pear-growing areas of Europe. The pathogen is able to colonize pear leaf debris and dead tissues of herbaceous plants and produce abundant ascospores and conidia that are capable of infecting pear trees. Inoculum monitoring in pear orchards is mainly achieved through spore traps and species identification is based on conidial morphology, but the similarities on conidial traits among species of Stemphylium make correct identification difficult. In this work a total of thirty-seven Stemphylium isolates from pear orchards were characterized at the morphological, pathogenic, and molecular level. Correspondence among ITS and gpd sequences and morphological traits were evaluated. Species identification based exclusively on morphological data was not feasible. Combined morphological and molecular data were necessary for unambiguous identification of isolates in the S. vesicarium species group. Only isolates identified as S. vesicarium were pathogenic on pear. The study revealed that several species of Stemphylium coexist in pear orchards with S. vesicarium, the causal agent of BSP, and that combined morphological and molecular data are needed to differentiate them. Consequently, direct measurements of the airborne inoculum using volumetric spore traps may overestimate the actual pathogen population.

  9. Control of postharvest storage rots of apples and pears in The Netherlands

    NARCIS (Netherlands)

    Wenneker, M.; Köhl, J.; Leeuwen, van P.J.; Pham, K.T.K.; Schaik, van A.C.R.

    2016-01-01

    Postharvest diseases are a major problem in long storage of apples and pears in
    The Netherlands. Despite intensive preharvest spraying programs significant losses occur. Over 150 heavily affected many apples (mainly ‘Elstar’) and pears (mainly ‘Conference’) from packinghouses in different region

  10. 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 ...

  11. 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 w

  12. Complete genome sequence of Japanese erwinia strain ejp617, a bacterial shoot blight pathogen of pear.

    Science.gov (United States)

    Park, Duck Hwan; Thapa, Shree Prasad; Choi, Beom-Soon; Kim, Won-Sik; Hur, Jang Hyun; Cho, Jun Mo; Lim, Jong-Sung; Choi, Ik-Young; Lim, Chun Keun

    2011-01-01

    The Japanese Erwinia strain Ejp617 is a plant pathogen that causes bacterial shoot blight of pear in Japan. Here, we report the complete genome sequence of strain Ejp617 isolated from Nashi pears in Japan to provide further valuable insight among related Erwinia species.

  13. 76 FR 4202 - Pears Grown in Oregon and Washington; Amendment To Allow Additional Exemptions

    Science.gov (United States)

    2011-01-25

    ... order regulates the handling of pears grown in Oregon and Washington. Local administration of the... action is intended to provide regulatory flexibility to small pear handlers, while facilitating the sale... INFORMATION: This rule is issued under Marketing Order No. 927, as amended (7 CFR part 927), regulating...

  14. Polyribosomes from Pear Fruit: Changes during Ripening and Senescence.

    Science.gov (United States)

    Drouet, A; Hartmann, C

    1979-12-01

    Polysome profiles were examined from lyophilized peel tissue of ripening pear (Pyrus communis, L. var. Passe-Crassane). Messenger RNA chains bearing up to eight ribosomes (octamers) were resolved and exhibited the highest absorption peak when ribonuclease activity was eliminated during extraction. Neither normal ripening nor the increase of large polyribosomes that normally accompanies ripening and senescence of the fruit occurred when pretreatment at 0 C was omitted. Normal ripening and increase of large polyribosomes would, however, be initiated by an ethylene treatment. The size distribution of the polyribosomes remained essentially constant throughout a 4-month cold storage; there was, however, a large increase in ribosomes by the 12th week of storage.

  15. 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.

  16. Hardness amplification in nondeterministic logspace

    OpenAIRE

    Gupta, Sushmita

    2007-01-01

    A hard problem is one which cannot be easily computed by efficient algorithms. Hardness amplification is a procedure which takes as input a problem of mild hardness and returns a problem of higher hardness. This is closely related to the task of decoding certain error-correcting codes. We show amplification from mild average case hardness to higher average case hardness for nondeterministic logspace and worst-to-average amplification for nondeterministic linspace. Finally we explore possible ...

  17. Kinetic Hairpin Oligonucleotide Blockers for Selective Amplification of Rare Mutations

    Science.gov (United States)

    Jia, Yanwei; Sanchez, J. Aquiles; Wangh, Lawrence J.

    2014-01-01

    Detection of rare mutant alleles in an excess of wild type alleles is increasingly important in cancer diagnosis. Several methods for selective amplification of a mutant allele via the polymerase chain reaction (PCR) have been reported, but each of these methods has its own limitations. A common problem is that Taq DNA polymerase errors early during amplification generate false positive mutations which also accumulate exponentially. In this paper, we described a novel method using hairpin oligonucleotide blockers that can selectively inhibit the amplification of wild type DNA during LATE-PCR amplification. LATE-PCR generates double-stranded DNA exponentially followed by linear amplification of single-stranded DNA. The efficiency of the blocker is optimized by adjusting the LATE-PCR temperature cycling profile. We also demonstrate that it is possible to minimize false positive signals caused by Taq DNA polymerase errors by using a mismatched excess primer plus a modified PCR profile to preferentially enrich for mutant target sequences prior to the start of the exponential phase of LATE-PCR amplification. In combination these procedures permit amplification of specific KRAS mutations in the presence of more than 10,000 fold excess of wild type DNA without false positive signals. PMID:25082368

  18. Comparison of recombinase-aid amplification and traditional polymerase chain reaction in DNA methylation detection of thyroid cancer%重组酶介导扩增技术与传统聚合酶链反应技术在甲状腺癌DNA甲基化检测中的应用比较

    Institute of Scientific and Technical Information of China (English)

    廖萍; 刘茶珍; 朱佩云; 刘国星; 王文静

    2013-01-01

    Objective To find a new technology and compare it with methylation specific polymerase chain reaction (MSP) in DNA methylation detection of thyroid cancer.Methods OXTR was selected as object of study.After samples DNA were extracted and were modified,the recombinase-aid amplification(RAA) and traditional MSP were separately used to amplify the target gene OXTR which was modified with bisulfite.Results Both the two technology succeeded in amplifying unmethylated OXTR gene.RAA succeeded in amplifying the methylated gene band.Conclusions RAA is a novel isothermal amplification technology in nucleic acid amplification technologies.It could be performed at 37 ℃ with no need to initial heat denaturation at a high temperature followed by amplification at a lower temperature,and this isothermal amplification technology may successfully compete with its widely used non-isothermal predecessor (PCR) for molecular biological study and application.%目的 建立重组酶介导的核酸扩增(RAA)技术特异性检测DNA甲基化的新方法并与传统的DNA甲基化特异性PCR(MSP)方法进行比较.方法 选取OXTR基因作为目的基因,提取样品外周血基因组DNA,经亚硫酸氢盐修饰后分别以MSP和RAA技术进行特异性检测DNA甲基化实验.结果 2种技术皆能扩增出OXTR非甲基化条带,而RAA技术成功扩增OXTR甲基化条带.结论 RAA是一种新型的等温体外核酸扩增技术,实现了在37℃恒温下的核酸快速扩增,可成为替代MSP乃至其他PCR实验的新方法.

  19. A continuum model for metabolic gas exchange in pear fruit.

    Directory of Open Access Journals (Sweden)

    Q Tri Ho

    2008-03-01

    Full Text Available Exchange of O(2 and CO(2 of plants with their environment is essential for metabolic processes such as photosynthesis and respiration. In some fruits such as pears, which are typically stored under a controlled atmosphere with reduced O(2 and increased CO(2 levels to extend their commercial storage life, anoxia may occur, eventually leading to physiological disorders. In this manuscript we have developed a mathematical model to predict the internal gas concentrations, including permeation, diffusion, and respiration and fermentation kinetics. Pear fruit has been selected as a case study. The model has been used to perform in silico experiments to evaluate the effect of, for example, fruit size or ambient gas concentration on internal O(2 and CO(2 levels. The model incorporates the actual shape of the fruit and was solved using fluid dynamics software. Environmental conditions such as temperature and gas composition have a large effect on the internal distribution of oxygen and carbon dioxide in fruit. Also, the fruit size has a considerable effect on local metabolic gas concentrations; hence, depending on the size, local anaerobic conditions may result, which eventually may lead to physiological disorders. The model developed in this manuscript is to our knowledge the most comprehensive model to date to simulate gas exchange in plant tissue. It can be used to evaluate the effect of environmental stresses on fruit via in silico experiments and may lead to commercial applications involving long-term storage of fruit under controlled atmospheres.

  20. 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.

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

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Based on the introduction of the natural and geographical 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 Fragrant Pear is introduced from the aspects of the development status of the storage industry the development status of processing industry,and the status of domestic and foreign marketing.Problems in the industrialization development of Korla Fragrant Pear in Bazhou are analyzed,such as the weak protection of brand and lack of external propaganda,the imperfect benefit affiliating mechanism between leading enterprises and peasant households,and the marketing network of Korla Fragrant Pear and single mode of marketing.Countermeasures for the acceleration of the industrialization development of Korla Fragrant Pear in Bazhou are put forward,such as making great effort at publicity,brand establishment and counterfeit prevention,cultivating leading enterprises,reducing market risk,implementing industrialization development,adopting various marketing forms and actively developing domestic and international markets.

  2. Betalain, Acid Ascorbic, Phenolic Contents and Antioxidant Properties of Purple, Red, Yellow and White Cactus Pears

    Science.gov (United States)

    Sumaya-Martínez, María Teresa; Cruz-Jaime, Sandra; Madrigal-Santillán, Eduardo; García-Paredes, Juan Diego; Cariño-Cortés, Raquel; Cruz-Cansino, Nelly; Valadez-Vega, Carmen; Martinez-Cardenas, Leonardo; Alanís-García, Ernesto

    2011-01-01

    Commercialization of cactus pears based on their antioxidant properties can generate competitive advantages, and these can turn into business opportunities and the development of new products and a high-value ingredient for the food industry. This work evaluated the antioxidant activities (1,1-diphenyl-2-picrylhydrazyl radical-scavenging, protection against oxidation of a β-carotene-linoleic acid emulsion, and iron (II) chelation), the content of total phenolic compounds, ascorbic acid, betacyanin, betaxanthin and the stability of betacyanin pigments in presence of Cu (II)-dependent hydroxyl radicals (OH•), in 18 cultivars of purple, red, yellow and white cactus pear from six Mexican states. Our results indicated that the antiradical activities from yellow and white cactus pear cultivars were not significantly different (p < 0.05) and were lower than the average antiradical activities in red and purple cultivars. The red cactus pear from the state of Zacatecas showed the highest antioxidant activity. The free radical scavenging activity for red cactus pears was significantly correlated (p < 0.05) to the concentration of total phenolic compounds (R2 = 0.90) and ascorbic acid (R2 = 0.86). All 18 cultivars of cactus pears studied showed significant chelating activity of ferrous ions. The red and purple cactus pears showed a great stability when exposed to OH•. PMID:22072899

  3. Betalain, Acid Ascorbic, Phenolic Contents and Antioxidant Properties of Purple, Red, Yellow and White Cactus Pears

    Directory of Open Access Journals (Sweden)

    Leonardo Martinez-Cardenas

    2011-09-01

    Full Text Available Commercialization of cactus pears based on their antioxidant properties can generate competitive advantages, and these can turn into business opportunities and the development of new products and a high-value ingredient for the food industry. This work evaluated the antioxidant activities (1,1-diphenyl-2-picrylhydrazyl radical-scavenging, protection against oxidation of a β-carotene-linoleic acid emulsion, and iron (II chelation, the content of total phenolic compounds, ascorbic acid, betacyanin, betaxanthin and the stability of betacyanin pigments in presence of Cu (II-dependent hydroxyl radicals (OH•, in 18 cultivars of purple, red, yellow and white cactus pear from six Mexican states. Our results indicated that the antiradical activities from yellow and white cactus pear cultivars were not significantly different (p < 0.05 and were lower than the average antiradical activities in red and purple cultivars. The red cactus pear from the state of Zacatecas showed the highest antioxidant activity. The free radical scavenging activity for red cactus pears was significantly correlated (p < 0.05 to the concentration of total phenolic compounds (R2 = 0.90 and ascorbic acid (R2 = 0.86. All 18 cultivars of cactus pears studied showed significant chelating activity of ferrous ions. The red and purple cactus pears showed a great stability when exposed to OH•.

  4. Betalain, Acid ascorbic, phenolic contents and antioxidant properties of purple, red, yellow and white cactus pears.

    Science.gov (United States)

    Sumaya-Martínez, María Teresa; Cruz-Jaime, Sandra; Madrigal-Santillán, Eduardo; García-Paredes, Juan Diego; Cariño-Cortés, Raquel; Cruz-Cansino, Nelly; Valadez-Vega, Carmen; Martinez-Cardenas, Leonardo; Alanís-García, Ernesto

    2011-01-01

    Commercialization of cactus pears based on their antioxidant properties can generate competitive advantages, and these can turn into business opportunities and the development of new products and a high-value ingredient for the food industry. This work evaluated the antioxidant activities (1,1-diphenyl-2-picrylhydrazyl radical-scavenging, protection against oxidation of a β-carotene-linoleic acid emulsion, and iron (II) chelation), the content of total phenolic compounds, ascorbic acid, betacyanin, betaxanthin and the stability of betacyanin pigments in presence of Cu (II)-dependent hydroxyl radicals (OH•), in 18 cultivars of purple, red, yellow and white cactus pear from six Mexican states. Our results indicated that the antiradical activities from yellow and white cactus pear cultivars were not significantly different (p cactus pear from the state of Zacatecas showed the highest antioxidant activity. The free radical scavenging activity for red cactus pears was significantly correlated (p cactus pears studied showed significant chelating activity of ferrous ions. The red and purple cactus pears showed a great stability when exposed to OH•.

  5. Quince 'CPP': new dwarf rootstock for pear trees on organic and high density planting

    Directory of Open Access Journals (Sweden)

    Renato Vasconcelos Botelho

    2012-06-01

    Full Text Available In Brazil, pear production presents the same incipient situation over the last 15 years, due mostly to low production technology. In this context, this study aimed to evaluate the development, growth and production of the pear tree cultivars Cascatense, Tenra and Hosui grafted on 'CPP' quince rootstock, using 'FT' pear as interstem. This trial was carried out in Guarapuava, State of Paraná, Southern region of Brazil, by five productive cycles. The pear trees were planted in September of 2004, spaced at 1.0 x 4.0 m (2,500 trees ha-1, trained to the modified central leader, on a Four-wire trellis, with drip irrigation and cultivated under organic production system. The following variables were evaluated: sprouting, anthesis, yield, fruit weight, soluble solids content, titratable acidity, pulp firmness, canopy area per plant and per hectare and trunk diameter. The pear tree cv. Tenra was outstanding most of the years for fruit yield, and, consequently, showed the highest accumulated yield over the period (51.6 t ha-1, followed by the cultivars Cascatense (39.7 t ha-1 and Hosui (18.7 t ha-1. All pear cultivars presented suitable physical-chemical characteristics for commercial purposes, with minimal average soluble solids content of 11% at harvest. The maximum canopy area per hectare was attained for cv. Cascatense (3063.2 m², that was considered insufficient for a high yield. These results suggest the needs for studies with higher density planting and other training systems, searching optimize canopy volume. One of the most limiting factors in the organic pear orchard was the incidence of pear dieback caused by Botriosphaeria dothidea, severe more often in pear trees cv. Hosui.

  6. 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)

  7. Population growth of carmine cochineal in giant cactus pear artificially infested on laboratory conditions

    OpenAIRE

    2009-01-01

    The carmine cochineal (Dactylopius opuntiae) is up today, the main pest of the giant cactus pear in the states of Pernambuco, Paraíba and Ceará. This research aimed to measure the population growth of D. opuntiae in cladodes of giant cactus pear infested in the laboratory conditios. Cladodes of giant cactus pear were artificially infested with colonies carmine cochineal. The experiment was initiated on 10/02/2009 and ended 10/03/2009. Shaped population growth is a function of time and infesta...

  8. Polyethersulfone improves isothermal nucleic acid amplification compared to current paper-based diagnostics.

    Science.gov (United States)

    Linnes, J C; Rodriguez, N M; Liu, L; Klapperich, C M

    2016-04-01

    Devices based on rapid, paper-based, isothermal nucleic acid amplification techniques have recently emerged with the potential to fill a growing need for highly sensitive point-of-care diagnostics throughout the world. As this field develops, such devices will require optimized materials that promote amplification and sample preparation. Herein, we systematically investigated isothermal nucleic acid amplification in materials currently used in rapid diagnostics (cellulose paper, glass fiber, and nitrocellulose) and two additional porous membranes with upstream sample preparation capabilities (polyethersulfone and polycarbonate). We compared amplification efficiency from four separate DNA and RNA targets (Bordetella pertussis, Chlamydia trachomatis, Neisseria gonorrhoeae, and Influenza A H1N1) within these materials using two different isothermal amplification schemes, helicase dependent amplification (tHDA) and loop-mediated isothermal amplification (LAMP), and traditional PCR. We found that the current paper-based diagnostic membranes inhibited nucleic acid amplification when compared to membrane-free controls; however, polyethersulfone allowed for efficient amplification in both LAMP and tHDA reactions. Further, observing the performance of traditional PCR amplification within these membranes was not predicative of their effects on in situ LAMP and tHDA. Polyethersulfone is a new material for paper-based nucleic acid amplification, yet provides an optimal support for rapid molecular diagnostics for point-of-care applications.

  9. 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

  10. Rapid Amplification and Detection of Foodborne Pathogenic Rotavirus by Continuous-flow Reverse Transcription-Polymerase Chain Reaction Integrated with Online Fluorescence Analysis%集在线荧光分析的连续流动反转录-聚合酶链式反应快速扩增检测食源致病性轮状病毒

    Institute of Scientific and Technical Information of China (English)

    章春笋; 李彧媛; 王海英

    2011-01-01

    采用含RNA反转录(Reverse transcription,RT)和在线荧光分析的连续流动聚合酶链式反应(Po1ymerase chain reaction,PCR)微流控技术检测食源致病性轮状病毒.此RT-PCR微流控装置以加热铜块组成恒温带,以聚四氟乙烯毛细管微通道为反应体系构建而成.采用循环水冷却退火区,此装置能获得低至31℃的退火温度,而且温度控制及其稳定性良好,因而能满足不同PCR的要求.为了充分体现PCR微流控技术的优越性,在线荧光分析被用于检测PCR产物.当流速为7.5 mm/s时,轮状病毒RNA的cDNA扩增和在线荧光分析能在约12 min完成(扩增约10 min,在线分析约2 min).在此集成化的RT-PCR微流控装置上,Ih可完成轮状病毒RNA的RT-PCR以及其扩增产物的在线荧光分析,样品检出限达到6.4×10(4)copies/μL.%A continuous-flow polymerase chain reaction (PCR) microfluidics integrated with reverse transcription (RT) of RNA and online fluorescence analysis has been introduced, which has been successfully applied for fast amplification and identification of foodborne pathogenic Rotavirus. On this continuous-flow RT-PCR device, the isothermal heating copper blocks provides the temperatures for RT-PCR, and the polytetrafluoroethylene (PTFE) capillary was used as the RT-PCR reaction channel. By using the cycling water to cool the annealing zone, the annealing temperature as low as 31 ℃ could be obtained for the presented device, where good temperature control and stability was also achieved. Therefore, this device can meet different PCR requirements. To make the best of the advantages of microfluidic PCR, the online fluorescence analysis of amplification products has been performed. When the flow rate of 7.5 mm/s was used, the amplification of cDNA synthesized from Rotavirus RNA and then the online fluorescence analysis of amplification products could be completed in about 12 min ( about 10 min for amplification and 2 min for analysis). By using

  11. A Study of the Distribution of Apple stem pitting virus in Tissues of Pear Tree sing In Situ Hybridization and In Situ RT-PCR

    Institute of Scientific and Technical Information of China (English)

    ZHAO Ying; LIU Na; NIU Jian-xin

    2009-01-01

    To understand the distribution of Apple stem pitting virus (ASPV) in tissues of pear tree and provide directly theorical and technical support for shoot tips detoxication, leaves and shoot tips of Korla pear were used as the materials, cDNA probe for ASPV was synthesized through RT-PCR reaction system using the unradioactive digoxigenin-labeled probe, and the specificity and sensitivity of probe were verified by blot hybridization method. Paraffin slice for in situ PCR and in situ hybridization was made and the location and distribution of ASPV RNA were detected in paraffin slices using in situ reverse transcription polymerase chain reaction, and the important factors which influenced the experiment results were optimized. ASPV mainly distributed in palisade tissue of mesophyll cells, external cortex of the tip, and the corresponding newborn vascular bundles. 20 min was the suitable digestive time for proteinase K. For the better amplication, RT reaction system should be above 0.2 U μL-1 and 0.4 mmol L-1 for RNasin and dNTPs respectively, 0.1-1.3 U μL-1 SuperScript Ⅱ, 0.6-0.8 μmol L-1 primer concentration, and above 0.5 U 100 μL-1 LA Taq DNA polymerase. The suitable annealing temperature in PCR reaction was 60℃ with 35 cycles. The apical meristem of 0.25 mm was the region of virus-free.

  12. Effect of ultrasound on survival and growth of Escherichia coli in cactus pear juice during storage

    Directory of Open Access Journals (Sweden)

    Nelly del Socorro Cruz-Cansino

    2016-06-01

    Full Text Available Abstract The aim of this study was to investigate the effectiveness of ultrasound as a conservation method for the inactivation of Escherichia coli inoculated into cactus pear juices (green and purple. Total soluble solids, pH, titratable acidity, and the kinetics of E. coli in cactus pear juices treated by ultrasound (60%, 70%, 80% and 90% amplitude levels for 1, 3 and 5 min were evaluated over 5 days. Total inactivation was observed in both fruit juices after 5 min of ultrasound treatment at most amplitude levels (with the exception of 60% and 80%. After one and two days of storage, the recovery of bacteria counts was observed in all cactus pear juices. Ultrasound treatment at 90% amplitude for 5 min resulted in non-detectable levels of E. coli in cactus pear juice for 2 days. The parameters of pH, titratable acidity and soluble solids were unaffected.

  13. Effect of ultrasound on survival and growth of Escherichia coli in cactus pear juice during storage.

    Science.gov (United States)

    Cruz-Cansino, Nelly Del Socorro; Reyes-Hernández, Isidro; Delgado-Olivares, Luis; Jaramillo-Bustos, Diana Pamela; Ariza-Ortega, José Alberto; Ramírez-Moreno, Esther

    2016-01-01

    The aim of this study was to investigate the effectiveness of ultrasound as a conservation method for the inactivation of Escherichia coli inoculated into cactus pear juices (green and purple). Total soluble solids, pH, titratable acidity, and the kinetics of E. coli in cactus pear juices treated by ultrasound (60%, 70%, 80% and 90% amplitude levels for 1, 3 and 5min) were evaluated over 5 days. Total inactivation was observed in both fruit juices after 5min of ultrasound treatment at most amplitude levels (with the exception of 60% and 80%). After one and two days of storage, the recovery of bacteria counts was observed in all cactus pear juices. Ultrasound treatment at 90% amplitude for 5min resulted in non-detectable levels of E. coli in cactus pear juice for 2 days. The parameters of pH, titratable acidity and soluble solids were unaffected.

  14. 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...

  15. 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.

  16. Pink discoloration of canned pears: role of procyanidin chemical depolymerization and procyanidin/cell wall interactions.

    Science.gov (United States)

    Le Bourvellec, Carine; Gouble, Barbara; Bureau, Sylvie; Loonis, Michèle; Plé, Yves; Renard, Catherine M G C

    2013-07-10

    After canning, pear pieces turn occasionally from whitish-beige to pink. Conditions were set up to obtain this discoloration systematically and investigate its mechanism. Canned pears showed a significantly lower L* coordinate compared with fresh pears, and the L* coordinate of canned pears decreased with decreasing pH. The values of the a* and b* coordinates increased significantly after processing, the increase being greater for the more acidic pH values, with corresponding redder colors. After canning, polyphenol concentrations decreased significantly, mainly due to loss of procyanidins. This supported the hypothesis of conversion of procyanidins to anthocyanin-like compounds. However, no soluble product was detected at 520 nm, the characteristic wavelength of anthocyanins. When purified procyanidins were treated at 95 °C at three different pH values (2.7, 3.3, and 4.0), procyanidin concentrations decreased after treatment, the more so as the pH was lower, and a pinkish color also appeared, attributed to tannin-anthocyanidin pigment. The pink color was bound to cell walls. Extraction of the neoformed pink entities was attempted by successive solvent extractions followed by cell wall degrading enzymes. The pink color persisted in the residues, and canned pears gave significantly higher amounts of residues after solvent and enzyme treatments than fresh pears. Procyanidins were the entities responsible for the appearance of pink discoloration. However, it seems that this pink discoloration also involved the formation of strong, probably covalent, bonds to the cell wall.

  17. The effects of plant cover on population of pear psylla (Cacopsylla pyricola and its predators

    Directory of Open Access Journals (Sweden)

    Mohammad Saeed Emami

    2017-01-01

    Full Text Available Cacopsylla pyricola (Förster, 1848 (Hemiptera: Psyllidae is a serious pest of pear in all pear growing areas. In the scope of an integrated pest management, a two consecutive years study was carried out to determine the effects of plant cover on pear psyllid population and its predators. Two treatments including plant cover and bare ground were applied in a randomized complete block design with three replicates. The sampling of the pest and its predators were done weekly by beating technique and leaf sampling. The data were subjected to analysis of variance (ANOVA. The results showed that plant cover had significant effect on the increase of predators on the trees (P < 0.001. The psyllid specialist predator, Anthocoris nemoralis (Fabricius, 1794, had the highest population among the pear psyllid predators (0.29 per sample. Plant cover had no significant effect on reducing the population of eggs, nymphs and adults of the pear psyllid. Despite the increase in the population of predators led by plant cover, lack of their effectiveness to reduce the pear psyllid population is discussed.

  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. 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)

  20. Exponential quadruplex priming amplification for DNA-based isothermal diagnostics.

    Science.gov (United States)

    Partskhaladze, Tamar; Taylor, Adam; Lomidze, Levan; Gvarjaladze, David; Kankia, Besik

    2015-02-01

    Polymerase chain reaction (PCR) is a method of choice for molecular diagnostics. However, PCR relies on thermal cycling, which is not compatible with the goals of point-of-care diagnostics. A simple strategy to turn PCR into an isothermal method would be to use specific primers, which upon polymerase elongation can self-dissociate from the primer-binding sites. We recently demonstrated that a monomolecular DNA quadruplex, GGGTGGGTGGGTGGG, meets these requirements, which led to the development of the linear versions of quadruplex priming amplification (QPA). Here we demonstrate exponential version of isothermal QPA, which allows an unprecedented 10(10)-fold amplification of DNA signal in less than 40 min.

  1. 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.

  2. 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.

  3. Sequencing on products of Oncomelania hupensis through simple sequence repeat anchored polymerase chain reaction amplification%湖北钉螺微卫星锚定PCR产物序列分析

    Institute of Scientific and Technical Information of China (English)

    郭俊涛; 周艺彪; 韦建国; 赵根明

    2008-01-01

    目的 分析4个种群湖北钉螺中微卫星序列及其两侧序列的特点.方法 以(CA)8RY为引物对湖北钉螺基因组DNA进行微卫星锚定PCR(SSR-PCR)扩增,对全部159个扩增产物进行T克隆并测定其中82个片段的核苷酸序列.结果 SSR-PCR的扩增产物是湖北钉螺基因组DNA上的散在区域,不是微卫星序列,但扩增产物中含有微卫星序列,测序的82个片段中36个克隆片段含有微卫星序列.微卫星序列其侧翼序列有一定的保守性,同一个微卫星的侧翼序列多数情况下是相同的.(GA/CT)n、(TTAGGG/CCCTAA)n两类微卫星在4个钉螺种群中均有发现,(CAA)n仅在福建福清种群中发现,(TCTCTG)n仅在安徽贵池种群中发现,(GAA~TTC)n、(CAA/TTG)n、(CAT)n三种微卫星序列仅在四川普格种群中发现.结论 SSR-PCR扩增的不是微卫星,其结果的分析应当类似于随机扩增多态性DNA.因此SSR-PCR不能十分有效体现微卫星作为分子标记的优势,应当根据微卫星的侧翼序列设计针对微卫星的引物,对微卫星进行PCR扩增并进行分析.%Objective To analyze the sequence of microsatellite and the flanking sequence from four populations of Oncomelania hupensis. Methods We cloned 159 SSR-PCR amplification products of a commonly used primer, (CA)8RY, using O. hupensis genomie DNA as template, and sequenced 82 products Results The sequences obtained were novel O. hupensis genomic sequences but not repeat simple sequence. It was observed that 36 out of 82 clones contained microsatellites between priming sites.The flanking sequences of certain microsatellite were invariant. Both (GA/CT). and (TTAGGG/CCCAA)n were found in four populations of O. hupensis. However, (CAA)n were found only in O. hupensis from Fuqing,Fujian province and (TCTCTG), were found only in O. hupensis from Guichi,Anhui province and (GAA/TTC)n, (CAA/TTG)n, (CAT), were found only in O.hupensis from Puge,Sichuan province. Conclusion The results obtained by

  4. Uncertainties in Site Amplification Estimation

    Science.gov (United States)

    Cramer, C. H.; Bonilla, F.; Hartzell, S.

    2004-12-01

    Typically geophysical profiles (layer thickness, velocity, density, Q) and dynamic soil properties (modulus and damping versus strain curves) are used with appropriate input ground motions in a soil response computer code to estimate site amplification. Uncertainties in observations can be used to generate a distribution of possible site amplifications. The biggest sources of uncertainty in site amplifications estimates are the uncertainties in (1) input ground motions, (2) shear-wave velocities (Vs), (3) dynamic soil properties, (4) soil response code used, and (5) dynamic pore pressure effects. A study of site amplification was conducted for the 1 km thick Mississippi embayment sediments beneath Memphis, Tennessee (see USGS OFR 04-1294 on the web). In this study, the first three sources of uncertainty resulted in a combined coefficient of variation of 10 to 60 percent. The choice of soil response computer program can lead to uncertainties in median estimates of +/- 50 percent. Dynamic pore pressure effects due to the passing of seismic waves in saturated soft sediments are normally not considered in site-amplification studies and can contribute further large uncertainties in site amplification estimates. The effects may range from dilatancy and high-frequency amplification (such as observed at some sites during the 1993 Kushiro-Oki, Japan and 2001 Nisqually, Washington earthquakes) or general soil failure and deamplification of ground motions (such as observed at Treasure Island during the 1989 Loma Prieta, California earthquake). Examples of two case studies using geotechnical data for downhole arrays in Kushiro, Japan and the Wildlife Refuge, California using one dynamic code, NOAH, will be presented as examples of modeling uncertainties associated with these effects. Additionally, an example of inversion for estimates of in-situ dilatancy-related geotechnical modeling parameters will be presented for the Kushiro, Japan site.

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

    DEFF Research Database (Denmark)

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

    2010-01-01

    is heat production. This can be converted to the rate of reaction and allows direct and continuous monitoring of the hydrolysis of complex substrates. To overcome the low molar enthalpy of the hydrolysis of the glycosidic bond, which is typically on the order of −2.5 kJ mol−1, an enzymatic signal......The study of cellulolytic enzymes has traditionally been carried out using endpoint measurements by quantitation of reaction products using high-performance liquid chromatography (HPLC) or overall determination of produced reducing ends. To measure catalytic activity, model substrates...... 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...

  6. 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

  7. 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, p

  8. Oligoribonucleotide (ORN) Interference-PCR (ORNi-PCR): A Simple Method for Suppressing PCR Amplification of Specific DNA Sequences Using ORNs

    OpenAIRE

    Naoki Tanigawa; Toshitsugu Fujita; Hodaka Fujii

    2014-01-01

    Polymerase chain reaction (PCR) amplification of multiple templates using common primers is used in a wide variety of molecular biological techniques. However, abundant templates sometimes obscure the amplification of minor species containing the same primer sequences. To overcome this challenge, we used oligoribonucleotides (ORNs) to inhibit amplification of undesired template sequences without affecting amplification of control sequences lacking complementarity to the ORNs. ORNs were effect...

  9. Magnetic Amplification by Magnetized Cosmic Rays in SNR Shocks

    CERN Document Server

    Riquelme, Mario A

    2009-01-01

    (Abridged) X-ray observations of synchrotron rims in supernova remnant (SNR) shocks show evidence of strong magnetic field amplification (a factor of ~100 between the upstream and downstream medium). This amplification may be due to plasma instabilities driven by shock-accelerated cosmic rays (CRs). One candidate is the cosmic ray current-driven (CRCD) instability (Bell 2004), caused by the electric current of large Larmor radii CRs propagating parallel to the upstream magnetic field. Particle-in-cell (PIC) simulations have shown that the back-reaction of the amplified field on CRs would limit the amplification factor of this instability to less than ~10 in galactic SNRs. In this paper, we study the possibility of further amplification driven near shocks by "magnetized" CRs, whose Larmor radii are smaller than the length scale of the field that was previously amplified by the CRCD instability. We find that additional amplification can occur due to a new instability, driven by the CR current perpendicular to t...

  10. Isothermal strand displacement amplification (iSDA): a rapid and sensitive method of nucleic acid amplification for point-of-care diagnosis.

    Science.gov (United States)

    Toley, Bhushan J; Covelli, Isabela; Belousov, Yevgeniy; Ramachandran, Sujatha; Kline, Enos; Scarr, Noah; Vermeulen, Nic; Mahoney, Walt; Lutz, Barry R; Yager, Paul

    2015-11-21

    We present a method of rapid isothermal amplification of DNA without initial heat denaturation of the template, and methods and probes for (a) real-time fluorescence detection and (b) lateral flow detection of amplicons. Isothermal strand displacement amplification (iSDA) can achieve >10(9)-fold amplification of the target sequence in isothermal DNA amplification methods. iSDA initiates at sites where DNA base pairs spontaneously open or transiently convert into Hoogsteen pairs, i.e. "breathe", and proceeds to exponential amplification by repeated nicking, extension, and displacement of single strands. We demonstrate successful iSDA amplification and lateral flow detection of 10 copies of a Staphylococcus aureus gene, NO.-inducible l-lactate dehydrogenase (ldh1) (Richardson, Libby, and Fang, Science, 2008, 319, 1672-1676), in a clean sample and 50 copies in the presence of high concentrations of genomic DNA and mucins in isothermal amplification reactions. Finally, we demonstrate the multiplexing capability of iSDA by the simultaneous amplification of the target gene and an engineered internal control sequence. The speed, sensitivity, and specificity of iSDA make it a powerful method for point-of-care molecular diagnosis.

  11. Adventitious shoot regeneration from the leaves of in vitro grown 'Zhongli 1' pear (Pyrus spp.)

    Institute of Scientific and Technical Information of China (English)

    Jie LIU; Xi ZHANG; Bharat Kumar POUDYAL; Yuxing ZHANG; Zhan JIAO; Jing QI

    2009-01-01

    The pear (Pyrus spp.) is one of the most important temperate fruit crops. The technique of adven-titious shoot regeneration from leaves is considered to be one of the shortcuts in the research on pear genetic modification and cellular engineering, which, however, has not been widely used. As the regeneration frequency of pear leaves is usually very low, the research on adventi-tious shoot regeneration from pear leaves is eagerly needed. In this experiment, the factors affecting shoot and bud regeneration from the leaves of 'Zhongli 1' pear were studied, and an efficient protocol for shoot regenera-tion was established. The results showed that different types of basic media, different combinations of plant growth regulators, leaf placement on medium, periods of dark culture and the use of silver nitrate (AgNO3) on culture media all significantly affected the adventitious shoot regeneration frequency of 'Zhongli 1' pear. The details are as follows: (1) Among three kinds of basic media, NN69 was better for 'Zhongli 1' shoot regenera- tion, followed by half(1/2) MS, while full MS had no effect on shoot regeneration; (2) Thidiazuron (TDZ) was better than 6-benzylaminopurine (6-BA) for 'Zhongli 1' regen-eration, with an optimal concentration of 1.5 mg.L-1, and the regeneration rate under this concentration could reach 85%, with 2.72 buds per leaf. 0.5 mg .L-1 indole-3-butyric acid (IBA), which induced a higher regeneration fre-quency, was a better choice for pear regeneration compared with 0.3 mg.L-1 naphthaleneacetic acid (NAA). Among the different combinations of plant growth regulators, TDZ + IBA was better for inducing high regeneration frequency; (3) The abaxial surface of leaves touching the medium was beneficial for leaves to uptake nutrients from the medium, and because of that, the regeneration fre-quency of leaves was significantly higher than that of leaves touching the medium with their adaxial surfaces (obverse side of leaf); (4) Dark culture was necessary

  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. 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.

  14. Comparative study of bioactive components in pear genotypes from Ardahan/Turkey

    Directory of Open Access Journals (Sweden)

    Zehra Tuğba Abacı

    2016-01-01

    Full Text Available In this study, 10 pear genotypes (İncir, Bal, Nene, Kabak, Banda, Kırmızı, İmlahor, Baraka, Limon and Güğüm, which grow in the Ardahan region, were evaluated for their total phenolic content, total anthocyanin content, brix°, pH, titratable acidity, total ascorbic acid content and antioxidant activity. According to the results, the pear genotypes used in this study had a high brix˚ content, high phenolic, anthocyanin and ascorbic acid contents, as well as high antioxidant activity. It was determined that ‘Bal’ pear had the highest total phenolic content and antioxidant activity. In ‘Nene’ and ‘Incir’ pears, the quantity of ascorbic acid and anthocyanin, as well as the antioxidant activity were less than those in the other genotypes. Correlations between brix° and pH, acidity and pH, peel phenolic content and flesh phenolic content, flesh ascorbic acid content and peel phenolic content, peel antioxidant activity and flesh phenolic content, were found to be significant. As a conclusion, due to the high levels of antioxidants and other bioactive compounds in pears, it is suggested to consume those fruits, especially with their peels. The results from this study will provide new insights into farming, fresh fruit consumption, industrial food processing and future research studies.

  15. 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.

  16. ATTRACTION OF MALE SUMMERFORM PEAR PSYLLA TO VOLATILES FROM FEMALE PSYLLA: EFFECTS OF FEMALE AGE, MATING STATUS, AND PRESENCE OF HOST PLANT

    Science.gov (United States)

    Pear psylla, Cacopsylla pyricola (Förster) (Hemiptera: Psyllidae), is a pest of pears throughout North America and western Europe. Previous studies in our laboratory showed that males of the overwintering form (winterform morphotype) were attracted to volatiles from pear shoots infested with post-d...

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

    Institute of Scientific and Technical Information of China (English)

    WANG Yong; GAO Zhaoming; XU Ying; LI Guangyu; HE Lisheng; QIAN Peiyuan

    2016-01-01

    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.

  18. Comparison of accessions from the UK and US national pear germplasm collections with a standardized set of microsatellite markers

    Science.gov (United States)

    A standardized set of 12 microsatellite markers, previously agreed upon following an ECP/GR workshop in 2006, was used to screen accessions from the UK National Pear Collection at Brogdale and from the US National Pear Germplasm Repository (NCGR), Corvallis. Eight standard varieties were chosen from...

  19. Isothermal Amplification of Insect DNA

    Science.gov (United States)

    The loop-mediated isothermal amplification of DNA (LAMP) method can amplify a target DNA sequence at a constant temperature in about one hour. LAMP has broad application in agriculture and medicine because of the need for rapid and inexpensive diagnoses. LAMP eliminates the need for temperature cycl...

  20. [Pretreatment method of near-infrared diffuse reflection spectra used for sugar content prediction of pears].

    Science.gov (United States)

    Wang, Wei-Ming; Dong, Da-Ming; Zheng, Wen-Gang; Zhao, Xian-De; Jiao, Lei-Zi; Wang, Ming-Fei

    2013-02-01

    The content of sugar is an important quality index for pears. However, the traditional sugar measurement methods are time-consuming and destructive. In the present study, the authors measured the sugar content of pears using visible and near infrared diffuse reflection spectroscopy. The pretreatment methods of multiplicative scatter correction (MSC), baseline correction, standard normal variate (SNV) transformation, and moving average algorithms were used on the original absorbance spectrum. Results indicate that the absorbance spectra after pretreatment are better than the original absorbance spectra for prediction. Partial least squares (PLS) regression was also used on the original absorbance spectrum and the absorbance spectrum after moving average and baseline correction. It follows that the forecast accuracy of the absorbance spectra after moving average is higher than that of the original absorbance spectra. The models gave good predictions of the sugar content of pears, with corresponding r values of 0.990 8, and standard errors of predictions of 0.019 0.

  1. 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....... 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....... to suffer from stress for water and nutrients due to the curtailed root systems, which may constrain fruit growth, reduce yield and quality. Thus, there is an urgent need to research on developing field strategies to mitigate those negative effects brought about by root pruning. The objective of the Ph...

  2. Ripening and in vitro retention of respiratory control by avocado and pear mitochondria.

    Science.gov (United States)

    Ozelkök, S I; Romani, R J

    1975-08-01

    The retention of respiratory control ("survival") by mitochondria held at 25 C was studied in relation to the ripening of two varieties of avocado (Persea americana Mill. var. ;Fuerte' and ;Hass') and one variety of pear (Pyrus communis. L. var. ;Bartlett') fruit. The survival of avocado mitochondria increased from 8 to 10 hours when isolated from unripe, preclimacteric fruit, to 48 hours when isolated from fully ripe, postclimacteric fruits. Although rates of alpha-ketoglutarate oxidation, respiratory control, and ADP/O decreased somewhat in the postclimacteric phase, survival per se was not affected. Pear mitochondria survived for more than 30 hours regardless of the physiological age of the source.Exposure of postclimacteric avocado mitochondria to a preclimacteric supernatant fraction curtailed their survival. The harmful effect of some unknown substance(s) in the preclimacteric avocado supernatant fraction was confirmed by utilizing pear mitochondria as an independent test system.

  3. Effect of salicylic acid (SA) on delaying fruit senescence of Huang Kum pear

    Institute of Scientific and Technical Information of China (English)

    ZHANG Yuxing; DU Guoqiang; WANG Guoying; ZHANG Jianghong; Hassan Imran

    2007-01-01

    This experiment was undertaken to explore the effect of salicylic acid (SA) at different concentrations on regulating fruit senescence ofHuang Kum pear.Through dipping fruits and fruit discs for a series of hours in SA solution,enzyme activities and physiological characteristics of Huang Kum pear were determined.The results revealed that SA enhanced the activity of superoxide dismutase (SOD) and peroxidase (POD) enzymes at 0.02 mmol/L and at 0.002 mmol/L with the treatment of dipping fruit discs for 4 h and 12 h,respectively.The malondialdehyde (MDA) contents were reduced at 0.002 mmol/L for 12 h,and water loss ratio was decreased at 0.5 mmol/L after 48 h of treatment.It was concluded that SA at lower concentrations could delay the senescence of Huang Kum pear fruit.

  4. Chemical and biochemical changes in prickly pears with different ripening behaviour.

    Science.gov (United States)

    Silos-Espino, Héctor; Fabian-Morales, Lourdes; Osuna-Castro, Juan Alberto; Valverde, María Elena; Guevara-Lara, Fidel; Paredes-López, Octavio

    2003-10-01

    Chemical and biochemical changes were studied in ripening prickly pears from three Opuntia morphospecies with different ripening behaviour: Naranjona (O. ficus-indica), Blanca Cristalina (Opuntia sp.), and Charola (O. streptacantha), of early, intermediate, and late ripening, respectively. At fullyripe stage (commercial maturity), Blanca Cristalina showed the biggest fruits, the hardest texture, and its pulp had the highest protein content. There were no significant differences among morphospecies in pH or total soluble solids in fully ripe fruits. The three species exhibited considerable levels of vitamin C, dietary fibre, and minerals such as calcium, iron, and zinc. Protein expression was analysed in pulp and skin from every species at physiological and commercial maturity. Some proteins appeared at both stages, while many others expressed differentially. This study evaluated prickly pear components important for human nutrition and health, and provided basic information on pricky pear ripening, with a view to its control and to improving shelf life.

  5. Maiden pear trees growth in replant soil after inoculation of rootstocks with mycorrhizal inoculum

    Directory of Open Access Journals (Sweden)

    Sławomir Świerczyński

    2015-03-01

    Full Text Available In the production of fruit trees it is important to set up a nursery always in a new site, alternatively to follow crop rotation rules. It is not always possible due to the size of a farm and production volume. In order to limit the effects of replant soil one can use different procedures before setting up a nursery. Chemical methods, however, must be replaced with non chemical ones due to environmental protection and reduction of production costs. In the experiment conducted in 2009-2012, growth of maiden pear trees of ‘Conference’ growing on three quince MA, MC, S1 rootstocks, cultivated in replant and non-replant soil after the use of mycorrhizal treatment, was compared. The strongest growth of maiden pear trees was obtained on non-replant soil with mycorrhizal and without mycorrhizal treatment of rootstocks. Inoculation of rootstocks influenced positively the height and fresh mass of the root system of maiden pear trees growing on two considered sites. On the other hand, inoculation did not rise the diameter of stem and number of lateral shoots of the maidens. Influence of mycorrhizal treatment of rootstocks on the length of lateral shoots was not obvious. Significantly the best results of maiden pear trees growth, except for the stem diameter, were obtained on MA quince compared to two other types. The mycorrhizal treatment gave better result of percentage obtained by maiden pear trees only in the replant site. The best efficiency of maiden pear trees in nursery production was observed for MA quince rootstock.

  6. Adaptive Responses of Birch-Leaved Pear (Pyrus betulaefolia Seedlings to Salinity Stress

    Directory of Open Access Journals (Sweden)

    Qiang Sheng WU

    2009-06-01

    Full Text Available One-year-old birch-leaved pear (Pyrus betulaefolia Bunge seedlings were subjected to 0, 50, 100, 150, and 200 mmol/L NaCl solutions for 27 days in order to study the effects of salinity stress on photosynthesis, ion accumulation and enzymatic and non-enzymatic scavenging of reactive oxygen species in the seedlings. The research was performed in a greenhouse using potted trees. Salinity stress reduced photosynthetic rates, stomatal conductance and water use efficiency of leaves of the pear seedlings, but increased transpiration rates and leaf temperature. Hydrogen peroxide and superoxide anion radical contents increased with increasing NaCl concentrations, a phenomena also observed for malondialdehyde, suggesting that leaves of the pear seedlings suffered from oxidative injury. Superoxide dismutase (SOD and catalase (CAT activities quickly responded by increasing when the pear seedlings were subjected to salinity stress. Total protein content in leaves of the seedlings was restrained by salinity stress, whereas ascorbate content increased. Salinity stress reduced glutathione content once the birch-leaved pear seedlings were exposed to a low level (50 or 100 mmol/L of NaCl, whereas a high level (150 or 200 mmol/L NaCl of salinity stress stimulated the accumulation of glutathione. Salinity stress increased the accumulation of Na+, Cl-, K+ and Mg2+ in the seedlings, but reduced Ca2+ levels and the ratio of other ions to Na+ except K+/Na+ under 50 mmol/L NaCl conditions. This suggests that leaves of birch-leaved pear seedlings possess the capacity for salt exclusion only under 50 mmol/L NaCl conditions, and Ca2+ does not play a fundamental role as a secondary messenger under salinity stress conditions.

  7. Strategies for Amplification of Trinucleotide Repeats: Optimization of Fragile X and Androgen Receptor PCR.

    Science.gov (United States)

    Papp; Snyder; Sedra; Guida; Prior

    1996-06-01

    Background: Trinucleotide repeat regions are heritable unstable elements that change in copy number from generation to generation. Amplification of these triplet repeats is an important diagnostic tool for molecular medicine. However, these repeats are often difficult to amplify and may require the use of different cosolvents or amplification strategies. Methods and Results: We used the fragile X and androgen receptor triplet repeat regions to demonstrate a series of conditions that may be used to optimize the amplification of repeat sequences. Conclusions: For androgen receptor, we show that predigestion of the template DNA was sufficient to generate consistent amplification. In the case of fragile X we found that predigestion, when combined with use of betaine as a destabilizing additive, was superior to other methods and yielded consistent amplification of normal and premutation alleles in both isotopic and nonisotopic reactions.

  8. Assessing Linearity of the Parasite Varroa destructor DNA Amplification

    Directory of Open Access Journals (Sweden)

    ODAGIU Antonia

    2009-12-01

    Full Text Available The importance of honeybee products make of disease prevention and control in honeybees one of the mainconcerns of beekeepers in the world. The PCR – RT reaction represents an alternative for amplification performed inorder to realize the Varroa destructor O. genotypization, very important stage in haoneybee resistance to parasitedescription and also in management of the treatments. The linearity data is a very important parameter and very usefulin determination of the amplification of the parasite DNA and success of the genotypization process. The amplificationefficiency was very satisfactory, fact revealed by the value of the regression line y = - 2.3103 * 26.552 together withcoefficient of determination equal (r2 = 0.9691, meaning that more than 96% of the reaction efficiency may beexplained by the process liniarity. The implementation of the RT-PCR method was successful and it represents apremise for validation process evolution.

  9. Performance of orange oil in the control of carmine cochineal in giant cactus pear.

    OpenAIRE

    2009-01-01

    Since its introduction, in 2001, the carmine cochineal (Dactylopius opuntiae) already decimated some 100.000 hectares of giant cactus pear (Opuntia ficus-indica) in semi-arid region of Paraiba. This study aimed to evaluate the behavior of five concentrations of orange oil, applied in cladodes on the death of D. opuntiae in field conditions. The research was carried out in a field of giant cactus pear infested by carmine cochineal on the site rigideira, Monteiro County, State of Paraíba. The ...

  10. Genotypic analysis of cutaneous T-cell lymphoma: a comparative study of Southern blot analysis with polymerase chain reaction amplification of the T-cell receptor-gamma gene.

    Science.gov (United States)

    Curcó, N; Servitje, O; Llucià, M; Bertran, J; Limón, A; Carmona, M; Romagosa, V; Peyrí, J

    1997-11-01

    The diagnosis of early cutaneous T-cell lymphoma (CTCL) is a difficult point in dermatology. Recently, Southern blot analysis (SBA) and polymerase chain reaction (PCR) have been used to detect clonality in initial lesions in which clinical and histological findings are unspecific. Forty-one samples from 25 patients with CTCL were investigated for the presence of T-cell receptor-gamma gene rearrangement using a nested PCR technique and analysed by polyacrylamide gel electrophoresis (PAGE). Conventional SBA was also performed on 28 samples from 20 of these patients. In addition, 20 samples corresponding to patients with large plaque parapsoriasis (LPP), cutaneous B-cell lymphoma (CBCL) and eczema were analysed by PCR in the same way as were the CTCL specimens. Most of the CTCL specimens (81%) showed clonality on PCR analysis. Among patients with mycosis fungoides, 71% of initial patch lesions and 100% of plaques and tumours showed clonal disease. Clonality could be detected in three of four histologically negative post-treatment lesions. Clonal rearrangement was detected in one of three patients with LPP and in three of 10 patients with CBCL. None of the samples corresponding to patients with eczema showed positive results. SBA was significantly less sensitive than PCR in detecting clonality in CTCL patients (42% among early disease and 60% among advanced cases). The results indicate that this PCR/PAGE technique is a reliable and useful method for the detection of clonality in early skin lesions of CTCL patients and probably in the identification of silent extracutaneous involvement.

  11. Shortening distance of forward and reverse primers for nucleic acid isothermal amplification.

    Science.gov (United States)

    Haitao, Qu; Wenchao, Zhang; Xiaohui, Zhang; Xiujun, Wang; Sulong, Li

    2014-06-01

    Existent nucleic acid isothermal detection techniques for clinical diseases are difficult to promote greatly due to limitations in such aspects as methodology, costs of detection, amplification efficiency and conditions for operation. There is therefore an urgent need for a new isothermal amplification method with the characteristics of high accuracy, easy operation, short time of detection and low costs. We have devised a new method of nucleic acid isothermal amplification using Bst DNA polymerase under isothermal conditions (60-65°C). We call this method of amplification by shortening the distance between forward and reverse primers for nucleic acid isothermal amplification SDAMP. The results demonstrated that this technique is highly sensitive, specific and has short reaction times (40-60 min). Results of sequencing show that the products of SDAMP amplification are mainly polymers formed by series connection of monomers formed through linkage of forward primer and complementary sequences in reverse primer via a few bases. The method is different from current methods of nucleic acid amplification. Our study shows, however, that it is a specific method of nucleic acid isothermal amplification depending on interactions between primers and DNA template.

  12. Ligation-Independent Mechanism of Multiplex Ligation-Dependent Probe Amplification

    OpenAIRE

    Uno, Naoki; Yanagihara, Katsunori

    2014-01-01

    Multiplex ligation-dependent probe amplification (MLPA) is a widely used technique for detecting genomic structural variants. The technique is based on hybridization and ligation, followed by amplification of the ligation products. Therefore, ligation is considered a fundamental process that determines the feasibility and fidelity of MLPA. However, despite the widespread use of this technique, its reaction mechanism has not been fully analyzed. Herein, we describe a ligation-independent pathw...

  13. PCR amplification of microsatellites from single cells of Karenia brevis preserved in Lugol's iodine solution.

    Science.gov (United States)

    Henrichs, D W; Renshaw, M A; Santamaria, C A; Richardson, B; Gold, J R; Campbell, L

    2008-01-01

    A simple and effective protocol is described for multiplex polymerase chain reaction (PCR) amplification of single cells of Karenia brevis. The protocol requires minimum processing, avoids additions that might dilute target DNA template, and can be used on cells preserved in Lugol's iodine preservative. Destaining of Lugol's-preserved cells with sodium thiosulfate allowed successful amplification of single-copy, nuclear-encoded microsatellites in single cells of K. brevis that have been preserved for up to 6 years.

  14. Spheromak Impedance and Current Amplification

    Energy Technology Data Exchange (ETDEWEB)

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

    2002-01-31

    It is shown that high current amplification can be achieved only by injecting helicity on the timescale for reconnection, {tau}{sub REC}, which determines the effective impedance of the spheromak. An approximate equation for current amplification is: dI{sub TOR}{sup 2}/dt {approx} I{sup 2}/{tau}{sub REC} - I{sub TOR}{sup 2}/{tau}{sub closed} where I is the gun current, I{sub TOR} is the spheromak toroidal current and {tau}{sub CLOSED} is the ohmic decay time of the spheromak. Achieving high current amplification, I{sub TOR} >> I, requires {tau}{sub REC} <<{tau}{sub 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 {tau}{sub 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 {proportional_to} I, or I{sub TOR} {approx} I. Program implications are discussed.

  15. Loop-mediated isothermal amplification of single pollen grains

    Institute of Scientific and Technical Information of China (English)

    Ali Bektaş; Ignacio Chapela

    2014-01-01

    The polymerase chain reaction (PCR) has been a reliable and fruitful method for many applications in ecology. Nevertheless, unavoidable technical and instrumental require-ments of PCR have limited its widespread application in field situations. The recent development of isothermal DNA amplifica-tion methods provides an alternative to PCR, which circumvents key limitations of PCR for direct amplification in the field. Being able to analyze DNA in the pol en cloud of an ecosystem would provide very useful ecological information, yet would require a field-enabled, high-throughput method for this potential to be realized. Here, we demonstrate the applicability of the loop-mediated DNA amplification method (LAMP), an isothermal DNA amplification technique, to be used in pol en analysis. We demonstrate that LAMP can provide a reliable method to identify species from the pol en cloud, and that it can amplify successful y with sensitivity down to single pol en grains, thus opening the possibility of field-based, high-throughput analysis.

  16. Research on the Formula of Reproduced Pear Vinegar%再制梨醋配方的研究

    Institute of Scientific and Technical Information of China (English)

    黄玉玲

    2014-01-01

    利用优质的梨为原料,经固态发酵获得梨醋,在此基础上,用梨汁、蜂蜜、糖调配成再制梨醋。再制梨醋与单纯的梨醋相比较,在口味上更柔和,更具水果的清香;营养更全面,是良好的佐餐调味品。主要研究了再制梨醋的加工技术与配方,由实验结果可以得到再制梨醋的最优配方组合:总酸度4 g/L,主要配料添加量:梨汁110 mL/L,蜂蜜2 g/L,糖7 g/L。%Take high-quality pears as raw materials,pear vinegar is obtained by solid state fermenta-tion,on this basis,blend pear j uice,honey and sugar to make pear vinegar.Compared to pure pear vinegar,the reproduced pear vinegar is with softer taste,more fruity fragrance,more comprehensive nutrition and it is a kind of good table seasoning.The processing technology and recipe of reproduced pear vinegar are studied in this paper.The experimental results show that the optimal formula of reproduced pear vinegar is:total acidity of 4 g/L,pear juice of 110 mL/L,honey of 2 g/L,sugar of 7 g/L.

  17. Diversity of unavailable polysaccharides and dietary fiber in domesticated nopalito and cactus pear fruit (Opuntia spp.).

    Science.gov (United States)

    Peña-Valdivia, Cecilia Beatriz; Trejo, Carlos; Arroyo-Peña, V Baruch; Sánchez Urdaneta, Adriana Beatriz; Balois Morales, Rosendo

    2012-08-01

    The aim of this study was to quantify mucilages, pectins, hemicelluloses, and cellulose of nopalitos (edible, as vegetable, young cladodes of flat-stemmed spiny cacti) of most consumed Mexican cultivars, and sweet and acid cactus pear fruits of Opuntia spp. The hypothesis is that, regardless of their unavailable polysaccharides diversity, nopalitos and cactus pear fruits are rich sources of soluble and insoluble dietary fiber. Twelve cultivars of Opuntia spp. were used. Nopalitos had a significant variation in structural polysaccharides among the cultivars: mucilages (from 3.8 to 8.6% dry matter (DM)) averaged near a half of pectins content (from 6.1 to 14.2% DM) and tightly bound hemicelluloses (from 2.2 to 4.7% DM), which were the less abundant polysaccharides, amounted 50% of the loosely bound hemicelluloses (from 4.3 to 10.7% DM). Acid fruits (or 'xoconostle') had significantly higher unavailable polysaccharides content than sweet fruit, and contain similar proportions than nopalitos. Unavailable polysaccharides represent a high proportion of dry tissues of nopalitos and cactus pear fruits, composition of both of these soluble and insoluble polysaccharides (total dietary fiber) widely vary among cultivars without an evident pattern. Nopalitos and cactus pear fruit can be considered an excellent source of dietary fiber.

  18. 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 ...

  19. 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.

  20. Characterization of the key aroma compounds in Bartlett pear brandies by means of the sensomics concept.

    Science.gov (United States)

    Willner, Bianca; Granvogl, Michael; Schieberle, Peter

    2013-10-09

    The aroma compounds in two commercial Bartlett pear brandies clearly differing in their overall aroma profiles were detected in the volatile fractions by the aroma extract dilution analysis. In brandy A eliciting the more intense pear-like, fruity aroma, ethyl (S)-2-methylbutanoate, (E)-β-damascenone, 1,1-diethoxyethane, 2- and 3-methylbutanol, (S)-2- and 3-methylbutanoic acid, and 2-phenylethanol were found with the highest Flavor Dilution (FD) factors. In brandy B judged to have a weaker overall aroma, also (E)-β-damascenone, ethyl (S)-2-methylbutanoate, and 2-phenylethanol revealed high FD factors, while many odorants showed lower FD factors. Fourty-four odor-active compounds were quantitated by stable isotope dilution assays, and the odor activity values (OAVs; ratio of concentrations to odor thresholds) confirmed (E)-β-damascenone and ethyl (S)-2-methylbutanoate as important aroma compounds in brandy A, while the OAVs of most odorants were much lower in brandy B. By aroma recombination studies, the aromas of both brandies could be matched using reference odorants in the same concentrations as they occurred in the spirits. In 15 commercial Bartlett pear brandies ethyl (E,Z)-2,4-decadienoate and (E,E)-2,4-decadienoate eliciting a pear-like aroma showed a reasonable correlation of their concentrations with the overall aroma quality.

  1. Genetics of biosynthesis and structure of the capsular exopolysaccharide from the Asian pear pathogen Erwinia pyrifoliae.

    Science.gov (United States)

    Kim, Won-Sik; Schollmeyer, Martin; Nimtz, Manfred; Wray, Victor; Geider, Klaus

    2002-12-01

    Erwinia pyrifoliae is a novel bacterial pathogen, which causes Asian pear blight and is related to Erwinia amylovora, the causative agent of fire blight. E. pyrifoliae produces exopolysaccharide (EPS) related to amylovoran in its sugar composition and sugar linkages. This was shown by degradation of the EPS with a viral depolymerase, and by methylation analysis and ESI/MS. The structure of the repeating units was confirmed by (1)H-NMR spectra. The EPS of E. pyrifoliae carried side chains, which were mainly terminated by acetyl and pyruvyl residues as found previously for amylovoran. On the other hand, a second side chain with glucose found for up to 65% of the repeating units of amylovoran was completely absent. The nucleotide sequences of five genes of the cps cluster of E. pyrifoliae encoding proteins for EPS synthesis were characterized and displayed a high homology with the corresponding ams genes. Similar functions of the gene products are assumed. As for ams mutants of E. amylovora, a cpsB mutant of E. pyrifoliae did not synthesize EPS and did not produce ooze on slices of immature pears or symptoms on pear seedlings. The cps mutant was complemented for EPS synthesis and virulence on pear slices with a gene cluster of E. amylovora that included amsB.

  2. 雪梨柠檬汁的研制%Preparation on pear lemon complex juice

    Institute of Scientific and Technical Information of China (English)

    梁巧荣; 植中强; 陆思敏

    2012-01-01

    选用雪梨、柠檬为原料研制雪梨柠檬复合饮料,将雪梨、柠檬分别榨汁,按一定的比例混合。雪梨柠檬汁的优化配方为:柠檬汁4%,雪梨汁20%,添加蜂蜜10%,复合稳定剂为0.1%CMC—Na+O.1%黄原胶。经调配、均质、灭菌等工艺制成色泽鲜艳、营养丰富、酸甜适口、老少皆宜的复合饮料。%Selection of pear and lemon as raw materials to prepare compound fruit drink. Respectively squeeze pear and lemon juice, then press the certain proportion mix. Pear lemon juice optimization formula is lemon juice 4%, pear juice 20%, honey 10%, composite stabilizers for 0.1% CMC-Na+0.1% xanthan gum. After deployment, homogeneous, sterilization and other processes into the brightly-colored,nutrient-rich, sweet and sour drink compound all ages.

  3. 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.

  4. 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...

  5. 75 FR 77563 - Nectarines, Pears, and Peaches Grown in California; Continuance Referenda

    Science.gov (United States)

    2010-12-13

    ... an effective means for determining whether growers favor the continuation of marketing order programs... Agricultural Marketing Service 7 CFR Parts 916 and 917 Nectarines, Pears, and Peaches Grown in California; Continuance Referenda AGENCY: Agricultural Marketing Service, USDA. ACTION: Referenda order. SUMMARY:...

  6. 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...

  7. Evaluation of fruit quality and susceptibility to blue mold of nine Asian pear cultivars

    Science.gov (United States)

    Nine Asian pear cultivars (Atago, Hosui, Isiiwase, Kosui, Olympic, Shinko, Shinsui, Ya Li, and Yoinashi) were evaluated for quality (firmness, titratable acidity, and soluble solids) and susceptibility to the blue mold pathogen Penicillium expansum. Fruit were grown at the University of Maryland Ext...

  8. 76 FR 8917 - Pears Grown in Oregon and Washington; Continuance Referendum

    Science.gov (United States)

    2011-02-16

    ...; ] DEPARTMENT OF AGRICULTURE Agricultural Marketing Service 7 CFR Part 927 Pears Grown in Oregon and Washington; Continuance Referendum AGENCY: Agricultural Marketing Service, USDA. ACTION: Referendum order. SUMMARY: This... referred to as the ``order,'' and the applicable provisions of the Agricultural Marketing Agreement Act...

  9. 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.

  10. Small Sample Whole-Genome Amplification

    Energy Technology Data Exchange (ETDEWEB)

    Hara, C A; Nguyen, C P; Wheeler, E K; Sorensen, K J; Arroyo, E S; Vrankovich, G P; Christian, A T

    2005-09-20

    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.

  11. 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

  12. 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.

  13. The draft genome sequence of European pear (Pyrus communis L. 'Bartlett').

    Science.gov (United States)

    Chagné, David; Crowhurst, Ross N; Pindo, Massimo; Thrimawithana, Amali; Deng, Cecilia; Ireland, Hilary; Fiers, Mark; Dzierzon, Helge; Cestaro, Alessandro; Fontana, Paolo; Bianco, Luca; Lu, Ashley; Storey, Roy; Knäbel, Mareike; Saeed, Munazza; Montanari, Sara; Kim, Yoon Kyeong; Nicolini, Daniela; Larger, Simone; Stefani, Erika; Allan, Andrew C; Bowen, Judith; Harvey, Isaac; Johnston, Jason; Malnoy, Mickael; Troggio, Michela; Perchepied, Laure; Sawyer, Greg; Wiedow, Claudia; Won, Kyungho; Viola, Roberto; Hellens, Roger P; Brewer, Lester; Bus, Vincent G M; Schaffer, Robert J; Gardiner, Susan E; Velasco, Riccardo

    2014-01-01

    We present a draft assembly of the genome of European pear (Pyrus communis) 'Bartlett'. Our assembly was developed employing second generation sequencing technology (Roche 454), from single-end, 2 kb, and 7 kb insert paired-end reads using Newbler (version 2.7). It contains 142,083 scaffolds greater than 499 bases (maximum scaffold length of 1.2 Mb) and covers a total of 577.3 Mb, representing most of the expected 600 Mb Pyrus genome. A total of 829,823 putative single nucleotide polymorphisms (SNPs) were detected using re-sequencing of 'Louise Bonne de Jersey' and 'Old Home'. A total of 2,279 genetically mapped SNP markers anchor 171 Mb of the assembled genome. Ab initio gene prediction combined with prediction based on homology searching detected 43,419 putative gene models. Of these, 1219 proteins (556 clusters) are unique to European pear compared to 12 other sequenced plant genomes. Analysis of the expansin gene family provided an example of the quality of the gene prediction and an insight into the relationships among one class of cell wall related genes that control fruit softening in both European pear and apple (Malus × domestica). The 'Bartlett' genome assembly v1.0 (http://www.rosaceae.org/species/pyrus/pyrus_communis/genome_v1.0) is an invaluable tool for identifying the genetic control of key horticultural traits in pear and will enable the wide application of marker-assisted and genomic selection that will enhance the speed and efficiency of pear cultivar development.

  14. The draft genome sequence of European pear (Pyrus communis L. 'Bartlett'.

    Directory of Open Access Journals (Sweden)

    David Chagné

    Full Text Available We present a draft assembly of the genome of European pear (Pyrus communis 'Bartlett'. Our assembly was developed employing second generation sequencing technology (Roche 454, from single-end, 2 kb, and 7 kb insert paired-end reads using Newbler (version 2.7. It contains 142,083 scaffolds greater than 499 bases (maximum scaffold length of 1.2 Mb and covers a total of 577.3 Mb, representing most of the expected 600 Mb Pyrus genome. A total of 829,823 putative single nucleotide polymorphisms (SNPs were detected using re-sequencing of 'Louise Bonne de Jersey' and 'Old Home'. A total of 2,279 genetically mapped SNP markers anchor 171 Mb of the assembled genome. Ab initio gene prediction combined with prediction based on homology searching detected 43,419 putative gene models. Of these, 1219 proteins (556 clusters are unique to European pear compared to 12 other sequenced plant genomes. Analysis of the expansin gene family provided an example of the quality of the gene prediction and an insight into the relationships among one class of cell wall related genes that control fruit softening in both European pear and apple (Malus × domestica. The 'Bartlett' genome assembly v1.0 (http://www.rosaceae.org/species/pyrus/pyrus_communis/genome_v1.0 is an invaluable tool for identifying the genetic control of key horticultural traits in pear and will enable the wide application of marker-assisted and genomic selection that will enhance the speed and efficiency of pear cultivar development.

  15. Application of Exogenous Ethylene Inhibits Postharvest Peel Browning of ‘Huangguan’ Pear

    Science.gov (United States)

    Ma, Yurong; Yang, Mengnan; Wang, Jingjing; Jiang, Cai-Zhong; Wang, Qingguo

    2017-01-01

    Peel browning disorder has an enormous impact on the exterior quality of ‘Huangguan’ pear whereas the underlying mechanism is still unclear. Although different methods have been applied for inhibiting the peel browning of ‘Huangguan’ pear, there are numerous issues associated with these approaches, such as time cost, efficacy, safety and stability. In this study, to develop a rapid, efficient and safe way to protect ‘Huangguan’ pear from skin browning, the effect of exogenous ethylene on peel browning of pear fruits stored at 0°C was evaluated. Results showed that ethylene treatments at 0.70–1.28 μL/L significantly decreased the browning rate and browning index from 73.80% and 0.30 to 6.80% and 0.02 after 20 days storage at 0°C, respectively, whereas ethylene treatments at 5 μL/L completely inhibited the occurrence of browning. In addition, ethylene treatments at 5 μL/L decreased the electrolyte leakage and respiration rate, delayed the loss of total phenolic compounds. Furthermore, ethylene (5 μL/L) treatment significantly enhanced the activity of catalase (CAT), ascorbate peroxidase (APX) and superoxide dismutase (SOD) and increased the 1, 1-diphenyl-2-picrylhydrazyl inhibition rate, but inhibited the activity of polyphenol oxidase (PPO) and peroxidase (POD). Our data revealed that ethylene prevented the peel browning through improving antioxidant enzymes (CAT, APX and SOD) activities and reducing PPO activity, electrolyte leakage rate and respiration rate. This study demonstrates that exogenous ethylene application may provide a safe and effective alternative method for controlling browning, and contributes to the understanding of peel browning of ‘Huangguan’ pear. PMID:28149298

  16. Biological and Pomological Characteristics of some Pear Varieties in Republic of Macedonia

    Directory of Open Access Journals (Sweden)

    Marjan Kiprjanovski

    2009-06-01

    Full Text Available Five years research of the biological and pomological characteristics of some pear varieties are presented in this paper. 18 pear varieties with different ripening time were included in the research. Some of investigated varieties are new bred (Turandot, Norma, Karmen, Monica, some of them are famous as resistant to Fire Blight (‘Harrow Delight’, ‘Harvest Queen’, ‘Harrow Sweet’, ‘Honey Sweet’ and some of them belong to group of old varieties that have been less investigated in our conditions (‘Starking Delicious’, ‘Abbe Fetel’, ‘Packams Triumph’, ‘Conference’, ‘Magness Highland’, ‘Guyot’. All varieties were compared with standard varieties (‘Williams’, ‘Precoce Moretini’, ‘Bosc,s’. Demonstrative orchard was established in 1997 at the Faculty farm-Trubarevo near Skopje. The trees were grafted on BA 29 quince rootstock and for those varieties that had not affinity with quince, we used inter stock pear variety Cure. Research was conducted during the period of 2003-2007. The following parameters were investigated: blossom, ripening of the fruits, growth of the trees (TCSA, volume of the crowns, yield, pomological characteristics of the fruits and content of the soluble dry matters and acids. The results showed that some of the pear varieties are characterized with a good production attributes, resistant to Fire Blight, pear sucker and to some a biotical factors, with good quality of the fruits. In the first group, the best characteristics gave variety Norma, from resistant variety group good results gave variety Harrow Sweet, and from old variety assortment for our conditions, good results gave ‘Packams Triumph’. These varieties can be recommended for cultivation in our climatically conditions.

  17. 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.

  18. DC-driven thermoelectric Peltier device for precise DNA amplification

    Science.gov (United States)

    Yamaguchi, Shigeo; Suzuki, Tadzunu; Inoue, Kazuhito; Azumi, Yoshitaka

    2015-05-01

    Using a DC-driven Peltier device, we fabricated a DNA amplification system [polymerase chain reaction (PCR) system] with the aim of increasing its speed and precision. The Peltier device had a well block sandwiched by Bi2Se0.37Te2.36 as an N-type thermoelectric material and Bi0.59Sb1.30Te3 as a P-type material. The well block was directly controlled by the electric current, leading to a high thermal response. Using the Peltier device with the well block, we performed thermal cycles of a PCR, and we demonstrated that our PCR system produces a smaller amount of nonspecific products for the genome DNA (gDNA) of Arabidopsis thaliana, leading to a more precise DNA amplification system.

  19. Retrieval and Amplification of DNA from Unstained Histopathological Sections

    Institute of Scientific and Technical Information of China (English)

    DonnaC.MONTAGUE; BeverlyD.LYN-COOK; 等

    1993-01-01

    Testing of compounds for carcinogenic potential in vivo involves various experimental designs.A few of these techniques are directed to demonstrate the genotoxicity and mutagenicity of the compound by histopathology.These changes shown by histochemical means include monoclonal antibody directed cellular markers.Development of the polymerase chain reaction technique(PCR)for amplification of DNA has facilitated the investigation of molecular events related to the formation of malignant neoplasms.We describe here a method for screening tissues for mutations of the H-ras gene using monoclonal antibodies directed toward normal and mutant p21 proteins.Formalin-fixed,paraffinembedded tissue sections are used to subsequently confirm the gene mutation by PCR amplification of the H-ras gene.The results indicated a successful application of this technique to demonstrate the presence of p21 oncoprotein in the tissues tested.

  20. 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

  1. Powerful Amplification Cascades of FRET-Based Two-Layer Nonenzymatic Nucleic Acid Circuits.

    Science.gov (United States)

    Quan, Ke; Huang, Jin; Yang, Xiaohai; Yang, Yanjing; Ying, Le; Wang, He; Xie, Nuli; Ou, Min; Wang, Kemin

    2016-06-07

    Nucleic acid circuits have played important roles in biological engineering and have increasingly attracted researchers' attention. They are primarily based on nucleic acid hybridizations and strand displacement reactions between nucleic acid probes of different lengths. Signal amplification schemes that do not rely on protein enzyme show great potential in analytical applications. While the single amplification circuit often achieves linear amplification that may not meet the need for detection of target in a very small amount, it is very necessary to construct cascade circuits that allow for larger amplification of inputs. Herein, we have successfully engineered powerful amplification cascades of FRET-based two-layer nonenzymatic nucleic acid circuits, in which the outputs of catalyzed hairpin assembly (CHA) activate hybridization chain reactions (HCR) circuits to induce repeated hybridization, allowing real-time monitoring of self-assembly process by FRET signal. The cascades can yield 50000-fold signal amplification with the help of the well-designed and high-quality nucleic acid circuit amplifiers. Subsequently, with coupling of structure-switching aptamer, as low as 200 pM adenosine is detected in buffer, as well as in human serum. To our knowledge, we have for the first time realized real-time monitoring adaptation of HCR to CHA circuits and achieved amplified detection of nucleic acids and small molecules with relatively high sensitivity.

  2. DAF optimization using Taguchi methods and the effect of thermal cycling parameters on DNA amplification.

    Science.gov (United States)

    Caetano-Anollés, G

    1998-09-01

    Taguchi methods, which are widely applied in industrial process design, were used to optimize DNA amplification finger-printing (DAF). Quadratic loss functions that penalize deviations from prediction values and L9 (3(4)) and L18 (3(8)) orthogonal arrays revealed effects and interactions of amplification reaction components and thermal cycling parameters. Analysis of variance (ANOVA) decomposed the contribution of individual factors to the experimental response (amplification yield and product number), while verification experiments established that optimum conditions were predictable, verifiable and reproducible. While several amplification components (primer, magnesium and enzyme) conditioned the amplification reaction, annealing temperature and time were the only important thermal cycling contributing factors. The Taguchi strategy defined a robust and transportable amplification protocol based on high annealing temperatures (typically 48 degrees C) and primer concentrations (typically 8 microM), which can be applied to the fingerprinting of a wide range of DNA templates of plant and fungal origin. The general strategy of robust experimental design holds potential as an optimization tool for other methods in molecular biology.

  3. Antimicrobial Activity of Xoconostle Pears (Opuntia matudae) against Escherichia coli O157:H7 in Laboratory Medium

    Science.gov (United States)

    Hayek, Saeed A.; Ibrahim, Salam A.

    2012-01-01

    The objective of this study was to investigate the antimicrobial activity of xoconostle pears (Opuntia matudae) against Escherichia coli O157:H7. Xoconostle pears were sliced, blended, and centrifuged. The supernatant was then filtered using a 0.45 μm filter to obtain direct extract. Direct extract of xoconostle pears was tested against four strains of E. coli O157:H7 in brain heart infusion (BHI) laboratory medium using growth over time and agar well diffusion assays. Our results showed that direct extract of xoconostle pears had a significant (P < 0.05) inhibitory effect at 4, 6, and 8% (v/v) concentrations and complete inhibitory effect at 10% (v/v) during 8 h of incubation at 37°C. Minimum inhibitory volume (MIV) was 400 μL mL−1 (v/v) and minimum lethal volume (MLV) was 650 μL mL−1 (v/v). The inhibitory effect of xoconostle pears found to be concentration dependent and not strain dependent. Thus, xoconostle pears extract has the potential to inhibit the growth of E. coli O157:H7 and could provide a natural means of controlling pathogenic contamination, thereby mitigating food safety risks. PMID:22934117

  4. Antimicrobial Activity of Xoconostle Pears (Opuntia matudae against Escherichia coli O157:H7 in Laboratory Medium

    Directory of Open Access Journals (Sweden)

    Saeed A. Hayek

    2012-01-01

    Full Text Available The objective of this study was to investigate the antimicrobial activity of xoconostle pears (Opuntia matudae against Escherichia coli O157:H7. Xoconostle pears were sliced, blended, and centrifuged. The supernatant was then filtered using a 0.45 μm filter to obtain direct extract. Direct extract of xoconostle pears was tested against four strains of E. coli O157:H7 in brain heart infusion (BHI laboratory medium using growth over time and agar well diffusion assays. Our results showed that direct extract of xoconostle pears had a significant (P<0.05 inhibitory effect at 4, 6, and 8% (v/v concentrations and complete inhibitory effect at 10% (v/v during 8 h of incubation at 37°C. Minimum inhibitory volume (MIV was 400 μL mL−1 (v/v and minimum lethal volume (MLV was 650 μL mL−1 (v/v. The inhibitory effect of xoconostle pears found to be concentration dependent and not strain dependent. Thus, xoconostle pears extract has the potential to inhibit the growth of E. coli O157:H7 and could provide a natural means of controlling pathogenic contamination, thereby mitigating food safety risks.

  5. Structural, evolutionary and functional analysis of the class III peroxidase gene family in Chinese pear (Pyrus bretschneideri

    Directory of Open Access Journals (Sweden)

    Yun Peng Cao

    2016-12-01

    Full Text Available Peroxidases (PRXs are widely existed in various organisms and could be divided into different types according to their structures and functions. Specifically, the Class III Peroxidase, a plant-specific multi-gene family, involves in many physiological processes, such as the metabolism of auxin, the extension and thickening of cell wall, as well as the formation of lignin. By searching the pear genome database, 94 non-redundant PRXs from Pyrus bretschneideri (PbPRXs were identified. Subsequently, analysis of phylogenetic relationships, gene structures, conserved motifs, and microsynteny was performed. These PbPRXs were unevenly distributed among 17 chromosomes of pear. In addition, 26 segmental duplication events but only one tandem duplication were occurred in these PbPRXs, implying segmental duplication was the main contributor to the expansion of the PbPRX family. By the Ka/Ks analysis, 26 out of 27 duplicated PbPRXs has experienced purifying selection. Twenty motifs were identified in PbPRXs based on the MEME analysis, eleven of which were enriched in pear. A total of 41 expressed genes were identified from ESTs of pear fruit. According to qRT-PCR, the expression trends of five PbPRXs in subgroup C were consistent with the change of lignin content during pear fruit development. So we inferred that the five PbPRXs were candidate genes involved in the lignin synthesis pathway. These results provided useful information for further researches of PRX genes in pear.

  6. 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.

  7. 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

    2013-06-25

    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.

  8. Analysis of the Chemical Constituents of the Essential Oil from Anli Pear

    Institute of Scientific and Technical Information of China (English)

    XIN Guang; LIU Chang-jiang; HOU Dong-yan; XU Lin

    2004-01-01

    The essential oil of the whole fruit and the peel of Anli pear (Pyrus Ussriensis Maxim.)grown in the western region of Liaoning Province was extracted through the hydrodistillationmethod, and was investigated by gas chromatography-mass spectrometry (GC-MS) technique.The yields of the essential oils of Anli pear whole fruit and the peel were 0.073 and0.36%, respectively. The identification of the volatile compounds was conducted throughthe commercial National Institute of Science and Technology (NIST) and Wiley massspectral search program, confirmed by comparing the retention indices with standardvalues of authentic samples. A total of 7 and 16 components were identified from theessential oils of the peel and the whole fruit, respectively. The predominant constituentof the two kinds of essential oils was butylated hydroxytoluene, which is a typicalantioxidant.

  9. 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.

  10. Efficient Dye-Sensitized Solar Cells Using Red Turnip and Purple Wild Sicilian Prickly Pear Fruits

    Directory of Open Access Journals (Sweden)

    Aldo Di Carlo

    2010-01-01

    Full Text Available Dye-sensitized solar cells (DSSCs were assembled by using the bougainvillea flowers, red turnip and the purple wild Sicilian prickly pear fruit juice extracts as natural sensitizers of TiO2 films. The yellow orange indicaxanthin and the red purple betacyanins are the main components in the cocktail of natural dyes obtained from these natural products. The best overall solar energy conversion efficiency of 1.7% was obtained, under AM 1.5 irradiation, with the red turnip extract, that showed a remarkable current density (Jsc = 9.5 mA/cm2 and a high IPCE value (65% at λ = 470 nm. Also the purple extract of the wild Sicilian prickly pear fruit showed interesting performances, with a Jsc of 9.4 mA/cm2, corresponding to a solar to electrical power conversion of 1.26%.

  11. 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.

  12. The effect of variety and location on cactus pear (Opuntia ficus-indica) fruit quality.

    Science.gov (United States)

    de Wit, Maryna; Nel, Philip; Osthoff, Gernot; Labuschagne, Maryke T

    2010-06-01

    Little is known about the performance of South African cactus pear varieties in different agro-ecological regions. Effects of locality on internal quality parameters of available cactus pear varieties were examined. With only one exception, no significant differences among the mean replication values for the different parameters between the different locations were observed. The differences between mean values for most individual parameters at the three localities were highly significant. Highly significant differences between the mean values for the measured characteristics were observed, not only among the locations (except for the pulp glucose values), but also for the influences of genotype and interaction between locality and genotype. Significant variations existed between mean values of the different characteristics between localities. Genotype x environmental interactions were noted. It was concluded that Meyers is the most appropriate cultivar for economical purposes in South Africa.

  13. 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.

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

    OpenAIRE

    Vergara, Cristina; Beatriz CANCINO-MADARIAGA; Andrés RAMÍREZ-SALVO; Sáenz, Carmen; Robert, Paz; Lutz, Mariane

    2015-01-01

    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 poly...

  15. Seed Dispersal in the Iberian Pear, Pyrus bourgaeana: A Role for Infrequent Mutualists

    OpenAIRE

    Fedriani, José M.; Delibes, M.

    2009-01-01

    Seed dispersal by animals is a key interaction, with effects on the population ecology and evolution of many plant lineages. Despite the fact that infrequent seed dispersers can potentially provide important services to plant populations, little attention has been paid so far to scarce mutualists. We assessed different aspects of quantity and quality of seed dispersal from fruit removal to seed germination in the Iberian pear, Pyrus bourgaeana, finding that fruit consumers markedly differed i...

  16. Potential assessment of genome-wide association study and genomic selection in Japanese pear Pyrus pyrifolia

    OpenAIRE

    Iwata, Hiroyoshi; Hayashi, Takeshi; Terakami, Shingo; Takada, Norio; Sawamura, Yutaka; Yamamoto, Toshiya

    2013-01-01

    Although the potential of marker-assisted selection (MAS) in fruit tree breeding has been reported, bi-parental QTL mapping before MAS has hindered the introduction of MAS to fruit tree breeding programs. Genome-wide association studies (GWAS) are an alternative to bi-parental QTL mapping in long-lived perennials. Selection based on genomic predictions of breeding values (genomic selection: GS) is another alternative for MAS. This study examined the potential of GWAS and GS in pear breeding w...

  17. Starch distribution in pear tree organs in relation to training systems, rootstocks and fruit quality

    OpenAIRE

    Mesa Juliani, Karen

    2015-01-01

    Starch is the main form in which plants store carbohydrates reserves, both in terms of amounts and distribution among different plant species. Carbohydrates are direct products of photosynthetic activity, and it is well know that yield efficiency and production are directly correlated to the amount of carbohydrates synthesized and how these are distributed among vegetative and reproductive organs. Nowadays, in pear trees, due to the modernization of orchards, through the introduction of new r...

  18. Fungi of the genus Penicillium on apples and pears during the storage period

    Directory of Open Access Journals (Sweden)

    H. Borecka

    2015-06-01

    Full Text Available Isolation of Penicilium fungi in 1391 cases confirmed their presence on and pathogenicity to apples and pears. Four species were the cause of rotting of the fruits: P. expansum, P. diversum, P. cyclopium and P. spinulosum. All those species may occur separately or coexist in the mould spots. P. expansum spores may infect fruit with injured skin, only if they have an additional source of sugar and nitrogen.

  19. Amplification of cellular oncogenes in solid tumors

    Directory of Open Access Journals (Sweden)

    Ozkan Bagci

    2015-01-01

    Full Text Available The term gene amplification refers to an increase in copy number of a gene. Upregulation of gene expression through amplification is a general mechanism to increase gene dosage. Oncogene amplifications have been shown in solid human cancers and they are often associated with progression of cancer. Defining oncogene amplification is useful since it is used as a prognostic marker in clinical oncology nowadays, especially v-erb-b2 avian erythroblastic leukemia viral oncogene homolog 2 (HER2 targeted agents are used in breast cancer patients with high level of HER2 overexpression as a therapeutic approach. However, patients without HER2 overexpression do not appear to benefit from these agents. We concluded that determination of oncogene amplification in solid tumors is an important factor in treatment of human cancers with many unknowns. We have referred to PubMed and some databases to prepare this article.

  20. Reasoned opinion on the setting of new MRLs for azinphos-methyl in apples and pears

    Directory of Open Access Journals (Sweden)

    European Food Safety Authority

    2014-01-01

    Full Text Available In accordance with Article 6 of Regulation (EC No 396/2005, Germany, hereafter referred to as the evaluating Member State (EMS, received an application from Exponent International Ltd., on behalf of Makhteshim Chemical Works Ltd., to set an import tolerance for the active substance azinphos-methyl in apples and pears. To accommodate for the reported use in Argentina, Germany proposed to raise the existing MRLs from the limit of quantification of 0.05 mg/kg to 0.5 mg/kg. Germany drafted an evaluation report in accordance with Article 8 of Regulation (EC No 396/2005 which was submitted to the European Commission and forwarded to EFSA. According to EFSA the available supervised residue trials on apples and pears are not adequate to derive a MRL proposal since they do not reflect the critical GAP reported for Argentina and because of other deficiencies. In conclusion, EFSA does not recommend the setting of the import tolerances to accommodate for the reported use of azinphos-methyl in apples and pears in Argentina.

  1. Iron homeostasis and fire blight susceptibility in transgenic pear plants overexpressing a pea ferritin gene.

    Science.gov (United States)

    Djennane, Samia; Cesbron, Colette; Sourice, Sophie; Cournol, Raphael; Dupuis, Fabrice; Eychenne, Magali; Loridon, Karine; Chevreau, Elisabeth

    2011-05-01

    The bacterial pathogen Erwinia amylovora causes the devastating disease known as fire blight in some rosaceous plants including apple and pear. One of the pathogenicity factors affecting fire blight development is the production of a siderophore, desferrioxamine, which overcomes the limiting conditions in plant tissues and also protects bacteria against active oxygen species. In this paper we examine the effect of an iron chelator protein encoded by the pea ferritin gene on the fire blight susceptibility of pear (Pyrus communis). Transgenic pear clones expressing this gene controlled either by the constitutive promoter CaMV 35S or by the inducible promoter sgd24 promoter were produced. The transgenic clones produced were analysed by Q-RT-PCR to determine the level of expression of the pea transgene. A pathogen-inducible pattern of expression of the pea transgene was observed in sgd24-promoter transformants. Adaptation to iron deficiency in vitro was tested in some transgenic clones and different iron metabolism parameters were measured. No strong effect on iron and chlorophyll content, root reductase activity and fire blight susceptibility was detected in the transgenic lines tested. No transformants showed a significant reduction in susceptibility to fire blight in greenhouse conditions when inoculated with E. amylovora.

  2. Chilling privation during dormancy period and carbohydrate mobilization in Japanese pear trees

    Directory of Open Access Journals (Sweden)

    Anderson Carlos Marafon

    2011-08-01

    Full Text Available The flower bud abortion is one of the main problems that limit commercial pear (Pyrus pyrifolia production in the southern region of Brazil. Insufficient chilling during the dormancy period is known as the main factor of this problem. One of the hypotheses to explain this problem is that the starch mobilization and carbohydrate fluxes to the buds are impeded when mild temperatures occurred during winter. This study compared the total soluble sugars (TSS and reducing sugars (RS concentrations, the cell wall acid invertase (CWAI - EC 3.2.1.26 and sucrose-phosphate synthase (SPS - EC 2.4.1.14 activities in wood of branches and floral buds of Japanese pear trees cv. Housui, grafted on Pyrus calleryana and submitted to chilling conditions during the dormancy period. Treatments were: (i natural conditions; (ii continuous artificial chilling; (iii alternating temperatures, and (iv total chilling privation. TSS and RS contents, as well as CWAI and SPS activities in tissues of branches that received insufficient chilling were lower than those that received sufficient chilling during winter. The starch concentration was superior in wood tissues of branches kept under chilling privation. The chilling privation disturbs carbohydrate mobilization in pear trees, reducing the sucrose synthesis capacity in wood tissues (source and sucrose importation by the floral buds (sink.

  3. Potential assessment of genome-wide association study and genomic selection in Japanese pear Pyrus pyrifolia.

    Science.gov (United States)

    Iwata, Hiroyoshi; Hayashi, Takeshi; Terakami, Shingo; Takada, Norio; Sawamura, Yutaka; Yamamoto, Toshiya

    2013-03-01

    Although the potential of marker-assisted selection (MAS) in fruit tree breeding has been reported, bi-parental QTL mapping before MAS has hindered the introduction of MAS to fruit tree breeding programs. Genome-wide association studies (GWAS) are an alternative to bi-parental QTL mapping in long-lived perennials. Selection based on genomic predictions of breeding values (genomic selection: GS) is another alternative for MAS. This study examined the potential of GWAS and GS in pear breeding with 76 Japanese pear cultivars to detect significant associations of 162 markers with nine agronomic traits. We applied multilocus Bayesian models accounting for ordinal categorical phenotypes for GWAS and GS model training. Significant associations were detected at harvest time, black spot resistance and the number of spurs and two of the associations were closely linked to known loci. Genome-wide predictions for GS were accurate at the highest level (0.75) in harvest time, at medium levels (0.38-0.61) in resistance to black spot, firmness of flesh, fruit shape in longitudinal section, fruit size, acid content and number of spurs and at low levels (pear.

  4. 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)

  5. Population growth of carmine cochineal in giant cactus pear artificially infested on laboratory conditions

    Directory of Open Access Journals (Sweden)

    Jacinto de Luna Batista

    2009-12-01

    Full Text Available The carmine cochineal (Dactylopius opuntiae is up today, the main pest of the giant cactus pear in the states of Pernambuco, Paraíba and Ceará. This research aimed to measure the population growth of D. opuntiae in cladodes of giant cactus pear infested in the laboratory conditios. Cladodes of giant cactus pear were artificially infested with colonies carmine cochineal. The experiment was initiated on 10/02/2009 and ended 10/03/2009. Shaped population growth is a function of time and infestation levels of initial and final, using a regression analysis with the application ASSISTAT 8.0 Beta. Data were also submitted to analysis of variance - ANOVA using a completely randomized design (CRD with eight treatments and five replications. The comparison of means was done by Tukey test at 5% probability. The results of the regression equations and curves showed that the insect Dactylopius opuntiae had a population growth in geometric progression in all treatments. Treatment eight colonies had the largest population growth where the average was obtained 1223.80 colonies / cladodes in 35 days. The lack of sunshine, average temperature of 22 º C and relative humidity of 75% RH during the study period, particularly favored the growth of the insect population.

  6. 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.

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

    Science.gov (United States)

    Poole, Catherine B; Tanner, Nathan A; Zhang, Yinhua; Evans, Thomas C; Carlow, Clotilde K S

    2012-01-01

    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.

  8. Pulse Compression And Raman Amplification In Optical Fibres

    Science.gov (United States)

    Byron, Kevin C.

    1988-06-01

    Experimental and theoretical investigations on Raman amplification in fibres have been carried out and simultaneous amplification and pulse compression observed. With a fibre design optimised for amplification high gain may be obtained at practical pump power levels.

  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. 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.

  11. Study on Enzymatic Characterization of Pyphenol Oxidase of Pear Fruit%蜜梨果实多酚氧化酶酶学特性的研究

    Institute of Scientific and Technical Information of China (English)

    邹礼根; 邱静; 赵芸; 姜慧燕; 刘军波

    2014-01-01

    多酚氧化酶(PPO)是酶促褐变的关键酶,与果蔬加工制品的色泽、抗氧化能力密切相关。以蜜梨果实为原料,邻苯二酚为底物,采用分光光度法研究蜜梨多酚氧化酶的酶学特性。结果表明:pH和温度对蜜梨PPO活性有明显的影响,其最适pH为4.5,最适温度为34℃。在加工过程中,可通过调节pH和温度来降低蜜梨PPO活性,减少褐变的发生;蜜梨PPO催化底物邻苯二酚的酶促反应动力学与米氏方程高度符合,R2=0.9972,其动力学方程为1V =0.17371[S]+0.4775,最大反应速率Vmax=2.09 U·min-1,米氏常数Km=0.36 mol·L-1;蜜梨PPO具有一定的热稳定性,随着温度的提高,完全抑制PPO活性所需要的时间逐渐减少。采用短时高温(90℃,1 min)的热处理,不仅可有效降低蜜梨加工过程中的酶促褐变,而且可减少蜜梨汁营养成分的损失,较好地保持其固有色泽。%Polyphenol oxidase (PPO) was the key enzyme of enzymatic browning, and was closely related to the color and antioxidant capacity of processed fruit and vegetable products. The enzymatic characterization of PPO from pears were studied in this paper with spectrophotometry method and based on substrate of catechol. The results showed that, both of pH and temperature had a significant effect on the activity of PPO of pear, the PPO had an optimum pH at 4.5 and optimum temperature at 34℃. In the process, the PPO activity of pear could be decreased by the regulation of pH and temperature to reduce the browning. The kinetics of the enzyme-catalyzed reaction of PPO was established and was in accord with Michaelis-Menten equation, and R2 was 0.997 2. The Km, Vmax and kinetic equation were respectively 0.36 mol·L-1, 2.09 U·min-1 and 1V =0.173 7 [1S]+0.477 5, using catechol as substrate. The PPO possessed certain thermal stability, furthermore, the higher the heating temperature was, the shorter time inhibiting PPO activity

  12. Linking Arctic amplification and local feedbacks

    Science.gov (United States)

    Balcerak, Ernie

    2011-11-01

    Climate simulations show that as the Earth warms, the Arctic warms more than the average global warming. However, models differ on how much more the Arctic warms, and although scientists have proposed a variety of mechanisms to explain the Arctic warming amplification, there is no consensus on the main reasons for it. To shed light on this issue, Hwang et al. investigated the relationship between Arctic amplification and poleward energy transport and local Arctic feedbacks, such as changes in cloud cover or ice loss, across a group of models. The researchers noted that differences in atmospheric energy transport did not explain the ranges of polar amplification; rather, models with more amplification showed less energy transport into high latitudes. The authors found that decreasing energy transport is due to a coupled relationship between Arctic amplification and energy transport: Arctic amplification reduces the equator-to-pole temperature gradient, which strongly decreases energy transport. They suggest that this coupled relationship should be taken into account in studies of Arctic amplification. (Geophysical Research Letters, doi:10.1029/2011GL048546, 2011)

  13. 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.

  14. 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.

  15. Molecular beacon probes combined with amplification by NASBA enable homogeneous, real-time detection of RNA

    NARCIS (Netherlands)

    Leone, G.; Schijndel, van H.; Gemen, van B.; Kramer, F.R.; Schoen, C.D.

    1998-01-01

    Molecular beacon probes can be employed in a NASBA amplicon detection system to generate a specific fluorescent signal concomitantly with amplification. A molecular beacon, designed to hybridize within the target sequence, was introduced into NASBA reactions that amplify the genomic RNA of potato le

  16. Critical evaluation of methods used to determine amplification efficiency refutes the exponential character of real-time PCR

    Directory of Open Access Journals (Sweden)

    Stewart Don

    2008-10-01

    Full Text Available Abstract Background The challenge of determining amplification efficiency has long been a predominant aspect of implementing real-time qPCR, playing a critical role in the accuracy and reliability that can be achieved. Based upon analysis of amplification profile position, standard curves are currently the gold standard for amplification efficiency determination. However, in addition to being highly resource intensive, the efficacy of this approach is limited by the necessary assumption that all samples are amplified with the same efficiency as predicted by a standard curve. These limitations have driven efforts to develop methods for determining amplification efficiency by analyzing the fluorescence readings from individual amplification reactions. The most prominent approach is based on analysis of the "log-linear region", founded upon the presumption that amplification efficiency is constant within this region. Nevertheless, a recently developed sigmoidal model has provided new insights that challenge such historically held views, dictating that amplification efficiency is not only dynamic, but is linearly coupled to amplicon DNA quantity. Called "linear regression of efficiency" or LRE, this kinetic-based approach redefines amplification efficiency as the maximal efficiency (Emax generated at the onset of thermocycling. Results This study presents a critical evaluation of amplification efficiency determination, which reveals that potentially large underestimations occur when exponential mathematics is applied to the log-linear region. This discrepancy was found to stem from misinterpreting the origin of the log-linear region, which is derived not from an invariant amplification efficiency, but rather from an exponential loss in amplification rate. In contrast, LRE analysis generated Emax estimates that correlated closely to that derived from a standard curve, despite the fact that standard curve analysis is founded upon exponential

  17. Quantum Amplitude Amplification and Estimation

    CERN Document Server

    Brassard, G; Mosca, M; Tapp, A; Brassard, Gilles; Hoyer, Peter; Mosca, Michele; Tapp, Alain

    2000-01-01

    Consider a Boolean function $\\chi: X \\to \\{0,1\\}$ that partitions set $X$ between its good and bad elements, where $x$ is good if $\\chi(x)=1$ and bad otherwise. Consider also a quantum algorithm $\\mathcal A$ such that $A \\ket{0} = \\sum_{x\\in X} \\alpha_x \\ket{x}$ is a quantum superposition of the elements of $X$, and let $a$ denote the probability that a good element is produced if $A \\ket{0}$ is measured. If we repeat the process of running $A$, measuring the output, and using $\\chi$ to check the validity of the result, we shall expect to repeat $1/a$ times on the average before a solution is found. *Amplitude amplification* is a process that allows to find a good $x$ after an expected number of applications of $A$ and its inverse which is proportional to $1/\\sqrt{a}$, assuming algorithm $A$ makes no measurements. This is a generalization of Grover's searching algorithm in which $A$ was restricted to producing an equal superposition of all members of $X$ and we had a promise that a single $x$ existed such tha...

  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. Chum-RNA allows preparation of a high-quality cDNA library from a single-cell quantity of mRNA without PCR amplification

    OpenAIRE

    2008-01-01

    Linear RNA amplification using T7 RNA polymerase is useful in genome-wide analysis of gene expression using DNA microarrays, but exponential amplification using polymerase chain reaction (PCR) is still required for cDNA library preparation from single-cell quantities of RNA. We have designed a small RNA molecule called chum-RNA that has enabled us to prepare a single-cell cDNA library after four rounds of T7-based linear amplification, without using PCR amplification. Chum-RNA drove cDNA synt...

  20. Chemical composition, in vitro gas production and energetic value of prickly pear fermented with and without Kluyveromyces marxianus

    Directory of Open Access Journals (Sweden)

    LESLIE BERUMEN

    2015-12-01

    Full Text Available The aim of this study was to characterize the chemical composition, in vitro gas production and energetic value of prickly pear during solid state fermentation (SSF with Kluyveromyces marxianus. Prickly pear was incubated in SSF at 28°C for 0, 24, 48, 72, 96 and 120 h without and inoculation with K. marxianus. The fermented cactus pear samples were dried at 55°C for 24 h, to determine the percent of dry matter (DM, crude protein (CP, neutral detergent fibre (NDF and acid detergent fibre (ADF. The volume of gas was recorded at 0, 3, 6, 9, 15, 24, 36, 48, 72 and 96 h and the parameters of in vitro gas production were obtained from the model: A= b*(1-e-c(t-L. In vitro gas production at 24 h was utilized for estimation of metabolizable energy (ME and short chain fatty acid (SCFA. Data were analyzed as a completely randomized design with a 2x6 factorial arrangement of treatments. The SSF in presence of K. marxianus increases the CP, maximum gas production (b, constant rate of gas production (c, ME and SCFA of prickly pear (P<0.05 but decreases NDF and ADF contents (P<0.05. We conclude that SSF with K. marxianus significantly improvement nutritional quality of prickly pear and may even promote animal performance.

  1. The Differences among Pear Genotypes to Fire Blight (Erwinia amylovora Attack, Based on Observations of Natural Infection

    Directory of Open Access Journals (Sweden)

    Adriana F. SESTRAS

    2008-08-01

    Full Text Available Fire blight, caused by the bacterium Erwinia amylovora, is one of the most damaging diseases of pear in the world. In Cluj-Napoca area, situated in central Transylvania, Romania, fire blight was observed first in 1994, very late comparative with the other countries from occidental Europe. The response of the pear cultivars and species from National Pear Collection from Cluj-Napoca to fire blight attack, assessed in natural conditions of infection, range on a large scale of variability, which denotes a strong influence of the genotype in expression of resistance or sensitivity to disease. From all genotypes, about 20.5% have not presented symptoms of attack, among them being the following: 'Blanquet precoce', 'Klementinka', 'Severianka', 'Beurre Bachelier', 'Kieffer Seedling', 'Er Shi Shinge', 'Beurre Amanlis', 'Bristol Cross', 'Beurre Liegel', 'Beurre Lucon', 'Grand Champion', 'Magness', 'Mericourt' etc. and several ancient autochthonous cultivars ('Pere malaiete', 'De zahar de Bihor', 'Cu miez rosu', 'Clopotele', 'Garoafa mare', 'Craiese', 'Para de apa'. Also, there were identified several species of Pyrus with no attack, as P. pollveria, P. common pear, P. lindlezi, P. malifolia, P. persica, P. ussuriensis, P. variolosa. The remarked genotypes could be potential sources for further breeding programmes and increase the number of genotypes available for breeding new pear cultivars resistant to Erwinia attack.

  2. Attraction of Male Winterform Pear Psylla to Female-produced Volatiles and to Female Extracts and Evidence of Male-Male Repellency

    Science.gov (United States)

    Pear psylla, Cacopsylla pyricola (Förster) (Hemiptera: Psyllidae), is a major pest of commercial pears in North America and Europe. Olfactometer trials have shown that males of both the summerform and winterform morphotype are attracted to female-infested host material. Additional work with the su...

  3. 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...

  4. Can Anomalous Amplification be Attained Without Postselection?

    CERN Document Server

    Martínez-Rincón, Julián; Viza, Gerardo I; Howell, John C

    2015-01-01

    We present a parameter estimation technique based on performing joint measurements of a weak interaction away from the weak-value-amplification approximation. Two detectors are used to collect full statistics of the correlations between two weakly entangled degrees of freedom. Without the need of postselection, the protocol resembles the anomalous amplification of an imaginary-weak-value-like response. The amplification is induced in the difference signal of both detectors allowing robustness to different sources of technical noise, and offering in addition the advantages of balanced signals for precision metrology. All of the Fisher information about the parameter of interest is collected, and a phase controls the amplification response. We experimentally demonstrate the proposed technique by measuring polarization rotations in a linearly polarized laser pulse. The effective sensitivity and precision of a split detector is increased when compared to a conventional continuous-wave balanced detection technique...

  5. 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.

  6. Rapid PCR amplification of DNA utilizing Coriolis effects.

    Science.gov (United States)

    Mårtensson, Gustaf; Skote, Martin; Malmqvist, Mats; Falk, Mats; Asp, Allan; Svanvik, Nicke; Johansson, Arne

    2006-08-01

    A novel polymerase chain reaction (PCR) method is presented that utilizes Coriolis and centrifugal effects, produced by rotation of the sample disc, in order to increase internal circulatory rates, and with them temperature homogenization and mixing speeds. A proof of concept has been presented by testing a rapid 45-cycle PCR DNA amplification protocol. During the repeated heating and cooling that constitutes a PCR process, the 100 microL samples were rotated at a speed equivalent to an effective acceleration of gravity of 7,000 g. A cycle time of 20.5 s gave a total process time of 15 min to complete the 45 cycles. A theoretical and numerical analysis of the resulting flow, which describes the increased mixing and temperature homogenization, is presented. The device gives excellent reaction speed efficiency, which is beneficial for rapid PCR.

  7. 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.

  8. Improved Performance of Loop-Mediated Isothermal Amplification Assays via Swarm Priming.

    Science.gov (United States)

    Martineau, Rhett L; Murray, Sarah A; Ci, Shufang; Gao, Weimin; Chao, Shih-Hui; Meldrum, Deirdre R

    2017-01-03

    This work describes an enhancement to the loop-mediated isothermal amplification (LAMP) reaction which results in improved performance. Enhancement is achieved by adding a new set of primers to conventional LAMP reactions. These primers are termed "swarm primers" based on their relatively high concentration and their ability to create new amplicons despite the theoretical lack of single-stranded annealing sites. The primers target a region upstream of the FIP/BIP primer recognition sequences on opposite strands, substantially overlapping F1/B1 sites. Thus, despite the addition of a new primer set to an already complex assay, no significant increase in assay complexity is incurred. Swarm priming is presented for three DNA templates: Lambda phage, Synechocystis sp. PCC 6803 rbcL gene, and human HFE. The results of adding swarm primers to conventional LAMP reactions include increased amplification speed, increased indicator contrast, and increased reaction products. For at least one template, minor improvements in assay repeatability are also shown. In addition, swarm priming is shown to be effective at increasing the reaction speed for RNA amplification via RT-LAMP. Collectively, these results suggest that the addition of swarm primers will likely benefit most if not all existing LAMP assays based on state-of-the-art, six-primer reactions.

  9. Flux variability scanning based on enforced objective flux for identifying gene amplification targets

    Directory of Open Access Journals (Sweden)

    Park Jong

    2012-08-01

    Full Text Available Abstract Background In order to reduce time and efforts to develop microbial strains with better capability of producing desired bioproducts, genome-scale metabolic simulations have proven useful in identifying gene knockout and amplification targets. Constraints-based flux analysis has successfully been employed for such simulation, but is limited in its ability to properly describe the complex nature of biological systems. Gene knockout simulations are relatively straightforward to implement, simply by constraining the flux values of the target reaction to zero, but the identification of reliable gene amplification targets is rather difficult. Here, we report a new algorithm which incorporates physiological data into a model to improve the model’s prediction capabilities and to capitalize on the relationships between genes and metabolic fluxes. Results We developed an algorithm, flux variability scanning based on enforced objective flux (FVSEOF with grouping reaction (GR constraints, in an effort to identify gene amplification targets by considering reactions that co-carry flux values based on physiological omics data via “GR constraints”. This method scans changes in the variabilities of metabolic fluxes in response to an artificially enforced objective flux of product formation. The gene amplification targets predicted using this method were validated by comparing the predicted effects with the previous experimental results obtained for the production of shikimic acid and putrescine in Escherichia coli. Moreover, new gene amplification targets for further enhancing putrescine production were validated through experiments involving the overexpression of each identified targeted gene under condition-controlled batch cultivation. Conclusions FVSEOF with GR constraints allows identification of gene amplification targets for metabolic engineering of microbial strains in order to enhance the production of desired bioproducts. The algorithm

  10. Identification of Self-Incompatibility Genotypes in Some Sand Pears (Pyrus pyrifolia Nakai) by PCR-RFLP Analysis

    Institute of Scientific and Technical Information of China (English)

    GU Qing-qing; ZHANG Qing-lin; HU Hong-ju; CHEN Qi-liang; LUO Zheng-rong

    2009-01-01

    The identification of self-incompatibility genotype (S-genotype) will be useful for selection of pollinizers and design of crossing in cultivar improvement of sand pear. This paper reported the identification of self-incompatibility genotypes of seven Chinese and two Japanese sand pear cultivars using PCR-RFLP analysis and S-Rnase sequencing. The S-genotypes of these cultivars were determined as follows: Huali 1 S1S3, Shounan S1S3, Xizili S1S4, Qingxiang S3S7, Sanhua S2S7, Huangmi (Imamuranatsu) S1S6, Huali 2 S3S4, Baozhuli S7S33, Cangxixueli S5S15. S-Rnase alleles (S1 to S9) in sand pear could be identified effectively by PCR-RFLP analysis.

  11. 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

  12. Improved monitoring of female codling moth (Lepidoptera: Tortricidae) with pear ester plus acetic acid in sex pheromone-treated orchards.

    Science.gov (United States)

    Knight, Alan

    2010-08-01

    The performance of clear delta traps baited with 3.0 mg of pear ester, ethyl (E,Z)-2,4-decadienoate, and 5.0 ml of acetic acid in separate lures was compared with orange delta traps baited with a single lure containing 3.0 mg of both pear ester and the sex pheromone, (E,E)-8,10-dodecadien-1-ol (codlemone) for codling moth, Cydia pomonella (L.), in apple, Malus domestica (Borkhausen). Residual analyses and field tests demonstrated that both the pear ester and acetic acid lures were effective for at least 8 wk. The two trap-lure combinations caught a similar number of total moths in an orchard treated with sex pheromone dispensers during short-term trials in 2008. However, the mean catch of female moths was significantly higher and male moths significantly lower in clear traps baited with pear ester and acetic acid versus orange traps baited with pear ester and codlemone. Season-long studies were conducted with these two trap-lure combinations in orchards treated with (n = 6) and without (n = 7) sex pheromone dispensers during 2009. The two trap-lure combinations caught similar numbers of moths in dispenser-treated orchards. In contrast, total catch was significantly higher (>2-fold) in the orange compared with the clear traps in untreated orchards. The clear caught >6-fold more females than the orange trap in both types of orchards. These studies suggest that deploying clear delta traps baited with pear ester and acetic acid can be an effective monitoring tool for female codling moth and an alternative to codlemone-baited traps in sex pheromone-treated orchards.

  13. 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.

  14. [Evaluation of Sugar Content of Huanghua Pear on Trees by Visible/Near Infrared Spectroscopy].

    Science.gov (United States)

    Liu, Hui-jun; Ying, Yi-bin

    2015-11-01

    A method of ambient light correction was proposed to evaluate the sugar content of Huanghua pears on tree by visible/near infrared diffuse reflectance spectroscopy (Vis/NIRS). Due to strong interference of ambient light, it was difficult to collect the efficient spectral of pears on tree. In the field, covering the fruits with a bag blocking ambient light can get better results, but the efficiency is fairly low, the instrument corrections of dark and reference spectra may help to reduce the error of the model, however, the interference of the ambient light cannot be eliminated effectively. In order to reduce the effect of ambient light, a shutter was attached to the front of probe. When opening shutter, the spot spectrum were obtained, on which instrument light and ambient light acted at the same time. While closing shutter, background spectra were obtained, on which only ambient light acted, then the ambient light spectra was subtracted from spot spectra. Prediction models were built using data on tree (before and after ambient light correction) and after harvesting by partial least square (PLS). The results of the correlation coefficient (R) are 0.1, 0.69, 0.924; the root mean square error of prediction (SEP) are 0. 89°Brix, 0.42°Brix, 0.27°Brix; ratio of standard deviation (SD) to SEP (RPD) are 0.79, 1.69, 2.58, respectively. The results indicate that, method of background correction used in the experiment can reduce the effect of ambient lighting on spectral acquisition of Huanghua pears in field, efficiently. This method can be used to collect the visible/near infrared spectrum of fruits in field, and may give full play to visible/near-infrared spectroscopy in preharvest management and maturity testing of fruits in the field.

  15. Effect of available nutrients on yield and quality of pear fruit Bartlett in Kashmir Valley India.

    Science.gov (United States)

    Dar, M A; Wani, J A; Raina, S K; Bhat, M Y; Dar, M A

    2012-11-01

    Pear is one of the most important commercial crops grown in the Kashmir valley of India. A study was conducted during 2008 to find out the effect of available nutrients on yield and quality parameters of pear cultivar "Bartlett" which revealed that nitrogen, phosphorus and potassium exhibited significant and positive relationship with fruit length (0.882, 0.856, and 0.482 mm, respectively), diameter (0.869, 0.794 and 0.458 mm, respectively), weight (0.876, 0.825 and 0.439 g, respectively), volume (0.908, 0.806 and 0.404, Cm3 respectively) and yield (0.908, 0.764 and 0.702 kg tree(-1), respectively) however, only nitrogen and phosphorus showed similar relationship with total sugars (0.833 and 0.838% respectively). The calcium indicated significant and negative relationship with fruit diameter (-0.433) and yield (-0.589), while as it showed significant and positive correlation with fruit firmness (0.442) only. The sulphur revealed significant and positive relationship with fruit length (0.440), diameter (0.434), TSS (0.482) and yield (0.729) whereas zinc, copper, iron and manganese exhibited significant and positive relationship with fruit length (0.889, 793, 0.671 and 0.619, respectively), diameter (0.875, 0.807, 0.653 and 0.576, respectively) weight (0.881, 0.784, 0.669 and 0.615, respectively), volume (0.885, 0.832, 0.692 and 0.572, respectively) TSS (0.858, 0.761, 0.735 and 0.609, respectively), total sugars (0.853, 0.890, 0.705 and 0.517, respectively) and yield (0.777, 0.618, 0.789 and 0.701, respectively). It is therefore suggested that nutrients have effect on quality and yield of pear fruits.

  16. [Hyperspectral technology combined with CARS algorithm to quantitatively determine the SSC in Korla fragrant pear].

    Science.gov (United States)

    Zhan, Bai-Shao; Ni, Jun-Hui; Li, Jun

    2014-10-01

    Hyperspectral imaging has large data volume and high dimensionality, and original spectra data includes a lot of noises and severe scattering. And, quality of acquired hyperspectral data can be influenced by non-monochromatic light, external stray light and temperature, which resulted in having some non-linear relationship between the acquired hyperspectral data and the predicted quality index. Therefore, the present study proposed that competitive adaptive reweighted sampling (CARS) algorithm is used to select the key variables from visible and near infrared hyperspectral data. The performance of CARS was compared with full spectra, successive projections algorithm (SPA), Monte Carlo-uninformative variable elimination (MC-UVE), genetic algorithm (GA) and GA-SPA (genetic algorithm-successive projections algorithm). Two hundred Korla fragrant pears were used as research object. SPXY algorithm was used to divided sample set to correction set with 150 samples and prediction set with 50 samples, respectively. Based on variables selected by different methods, linear PLS and nonlinear LS-SVM models were developed, respectively, and the performance of models was assessed using parameters r2, RMSEP and RPD. A comprehensive comparison found that GA, GA-SPA and CARS can effectively select the variables with strong and useful information. These methods can be used for selection of Vis-NIR hyperspectral data variables, particularly for CARS. LS-SVM model can obtain the best results for SSC prediction of Korla fragrant pear based on variables obtained from CARS method. r2, RMSEP and RPD were 0.851 2, 0.291 3 and 2.592 4, respectively. The study showed that CARS is an effectively hyperspectral variable selection method, and nonlinear LS-SVM model is more suitable than linear PLS model for quantitatively determining the quality of fra- grant pear based on hyperspectral information.

  17. Heat induces gene amplification in cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Yan, Bin, E-mail: yanbin@mercyhealth.com [Department of Radiation Oncology, University of Mississippi Medical Center, Jackson, MS 39213 (United States); Mercy Cancer Center, Mercy Medical Center-North Iowa, Mason City, IA 50401 (United States); Ouyang, Ruoyun [Department of Respiratory Medicine, The Second Xiangya Hospital, Xinagya School of Medicine, Central South University, Changsha 410011 (China); Huang, Chenghui [Department of Radiation Oncology, University of Mississippi Medical Center, Jackson, MS 39213 (United States); Department of Oncology, The Third Xiangya Hospital, Xinagya School of Medicine, Central South University, Changsha 410013 (China); Liu, Franklin [Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710 (United States); Neill, Daniel [Department of Radiation Oncology, University of Mississippi Medical Center, Jackson, MS 39213 (United States); Li, Chuanyuan [Dermatology, Duke University Medical Center, Durham, NC 27710 (United States); Dewhirst, Mark [Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710 (United States)

    2012-10-26

    Highlights: Black-Right-Pointing-Pointer This study discovered that heat exposure (hyperthermia) results in gene amplification in cancer cells. Black-Right-Pointing-Pointer Hyperthermia induces DNA double strand breaks. Black-Right-Pointing-Pointer DNA double strand breaks are considered to be required for the initiation of gene amplification. Black-Right-Pointing-Pointer The underlying mechanism of heat-induced gene amplification is generation of DNA double strand breaks. -- Abstract: Background: Hyperthermia plays an important role in cancer therapy. However, as with radiation, it can cause DNA damage and therefore genetic instability. We studied whether hyperthermia can induce gene amplification in cancer cells and explored potential underlying molecular mechanisms. Materials and methods: (1) Hyperthermia: HCT116 colon cancer cells received water-submerged heating treatment at 42 or 44 Degree-Sign C for 30 min; (2) gene amplification assay using N-(phosphoacetyl)-L-aspartate (PALA) selection of cabamyl-P-synthetase, aspartate transcarbarmylase, dihydro-orotase (cad) gene amplified cells; (3) southern blotting for confirmation of increased cad gene copies in PALA-resistant cells; (4) {gamma}H2AX immunostaining to detect {gamma}H2AX foci as an indication for DNA double strand breaks. Results: (1) Heat exposure at 42 or 44 Degree-Sign C for 30 min induces gene amplification. The frequency of cad gene amplification increased by 2.8 and 6.5 folds respectively; (2) heat exposure at both 42 and 44 Degree-Sign C for 30 min induces DNA double strand breaks in HCT116 cells as shown by {gamma}H2AX immunostaining. Conclusion: This study shows that heat exposure can induce gene amplification in cancer cells, likely through the generation of DNA double strand breaks, which are believed to be required for the initiation of gene amplification. This process may be promoted by heat when cellular proteins that are responsible for checkpoints, DNA replication, DNA repair and

  18. Bio-cultural anchorage of the prickly pear cactus in Tlalnepantla (Morelos), Mexico

    OpenAIRE

    Torres-Salcido, Gerardo; Ramos-Chávez, Alejandro; Urreta-Fernández, Álvaro

    2016-01-01

    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 Tla...

  19. 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.

  20. Training systems in Rocha pear. Evaluation of production, qualitative and economic aspects

    OpenAIRE

    Comporta, Ana Sofia Ferreira

    2010-01-01

    Mestrado em Engenharia Agronómica - Hortofruticultura e Viticultura - Instituto Superior de Agronomia With the aim of evaluating production, qualitative and economic aspects of four training systems during 2010, at Peral, Cadaval, 4-yr-old ‗Rocha‘ pear trees (Pyrus communis L.) grafted on Sydo and trained as vertical axis (1.0 x 4.0 m), open tatura trellis (0.8 x 4 m), palmette (1.2 x 4.0 m) and solaxe (1.0 x 4.0 m) were monitored. The highest canopy volume and the lower pruning weight wer...

  1. Training systems and prohexadione-calcium effect on vegetative growth and yield of Rocha pear

    OpenAIRE

    Ribeiro, Joana Sofia Santos Vidal

    2011-01-01

    Mestrado em Engenharia Agronómica - Instituto Superior de Agronomia Vegetative and reproductive aspects of 5 year old orchard of Rocha 'pear on Sydo rootstock trained in Vertical Axis, Palmette with 3 axes, Solaxe and Tatura systems were evaluated. Yield per hectare showed a large increase in 2011 compared with 2010, Vertical axis yield 88 tha-1, Palmette 66 tha-1, Solaxe 76 tha-1and Tatura 84 tha-1. Fruit thinning was not done. Palmette was the system whose branches had lower cross-sectio...

  2. 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

  3. One New Method of Nucleic Acid Amplification-Loop-mediated Isothermal Amplification of DNA

    Institute of Scientific and Technical Information of China (English)

    Xue-en FANG; Jian LI; Qin CHEN

    2008-01-01

    Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method, which amplifies DNA with high specificity, sensitivity, rapidity and efficiency under isothermal conditions using a set of four specially designed primers and a Bst DNA polymerase with strand displacement activity. The basic principle, characteristics, development of LAMP and its applications are summarized in this article.

  4. 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.

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

    Science.gov (United States)

    Fiore, Emmanuelle; Dausse, Eric; Dubouchaud, Hervé; Peyrin, Eric; Ravelet, Corinne

    2015-08-01

    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.

  6. Paralog-specific primers for the amplification of nuclear Loci in tetraploid barbels (barbus: cypriniformes).

    Science.gov (United States)

    Gante, Hugo F; Alves, Maria Judite; Dowling, Thomas E

    2011-01-01

    Thirty paralog-specific primers were developed, following an intron-primed exon-crossing strategy, for S7 and growth hormone genes in Barbus (subgenera Barbus and Luciobarbus). We found that paralog-specific amplification requires the use of only one paralog-specific primer, allowing their simultaneous use with universal exon-primed intron-crossing primers of broad taxonomic applicability. This hybrid annealing strategy guarantees both specificity and generality of amplification reactions and represents a step forward in the amplification of duplicated nuclear loci in polyploid organisms and members of multigene families. Assays of several representative taxa identified high levels of segregating single nucleotide polymorphisms (SNPs) and nucleotide diversity within each of these subgenera. Additionally, several insertions-deletions (indels) that are diagnostic across species are found in intronic regions. Therefore, these primers provide a reliable source of valuable nuclear SNP and indel data for population and species level studies of barbels, such as applied conservation and basic evolutionary studies.

  7. Determination of DQB1 alleles using PCR amplification and allele-specific primers.

    Science.gov (United States)

    Lepage, V; Ivanova, R; Loste, M N; Mallet, C; Douay, C; Naoumova, E; Charron, D

    1995-10-01

    Molecular genotyping of HLA class II genes is commonly carried out using polymerase chain reaction (PCR) in combination with sequence-specific oligotyping (PCR-SSO) or a combination of the PCR and restriction fragment length polymorphism methods (PCR-RFLP). However, the identification of the DQB1 type by PCR-SSO and PCR-RFLP is very time-consuming which is disadvantageous for the typing of cadaveric organ donors. We have developed a DQB1 typing method using PCR in combination with allele-specific amplification (PCR-ASA), which allows the identification of the 17 most frequent alleles in one step using seven amplification mixtures. PCR allele-specific amplification HLA-DQB1 typing is easy to perform, and the results are easy to interpret in routine clinical practice. The PCR-ASA method is therefore better suited to DQB1 typing for organ transplantation than other methods.

  8. An expeditious synthesis of spinasterol and schottenol, two phytosterols present in argan oil and in cactus pear seed oil, and evaluation of their biological activities on cells of the central nervous system.

    Science.gov (United States)

    Badreddine, Asmaa; Karym, El Mostafa; Zarrouk, Amira; Nury, Thomas; El Kharrassi, Youssef; Nasser, Boubker; Cherkaoui Malki, Mustapha; Lizard, Gérard; Samadi, Mohammad

    2015-07-01

    Spinasterol and schottenol, two phytosterols present in argan oil and in cactus pear seed oil, were synthesized from commercially available stigmasterol by a four steps reactions. In addition, the effects of these phytosterols on cell growth and mitochondrial activity were evaluated on 158N murine oligodendrocytes, C6 rat glioma cells, and SK-N-BE human neuronal cells with the crystal violet test and the MTT test, respectively. The effects of spinasterol and schottenol were compared with 7-ketocholesterol (7KC) and ferulic acid, which is also present in argan and cactus pear seed oil. Whatever the cells considered, dose dependent cytotoxic effects of 7KC were observed whereas no or slight effects of ferulic acid were found. With spinasterol and schottenol, no or slight effects on cell growth were detected. With spinasterol, reduced mitochondrial activities (30-50%) were found on 158N and C6 cells; no effect was found on SK-N-BE. With schottenol, reduced mitochondrial activity were revealed on 158N (50%) and C6 (10-20%) cells; no effect was found on SK-N-BE. Altogether, these data suggest that spinasterol and schottenol can modulate mitochondrial activity and might therefore influence cell metabolism.

  9. A novel thermostable polymerase for RNA and DNA Loop-mediated isothermal amplification (LAMP

    Directory of Open Access Journals (Sweden)

    Yogesh eChander

    2014-08-01

    Full Text Available Meeting the goal of providing point of care (POC tests for molecular detection of pathogens in low resource settings places stringent demands on all aspects of the technology. OmniAmp DNA polymerase (Pol is a thermostable viral enzyme that enables true POC use in clinics or in field by overcoming important barriers to isothermal amplification. In this paper, we describe the multiple advantages of OmniAmp Pol as an isothermal amplification enzyme and provide examples of its use in loop-mediated isothermal amplification (LAMP for pathogen detection. The inherent reverse transcriptase activity of OmniAmp Pol allows single enzyme detection of RNA targets in RT-LAMP. Common methods of nucleic acid amplification are highly susceptible to sample contaminants, necessitating elaborate nucleic acid purification protocols that are incompatible with POC or field use. OmniAmp Pol was found to be less inhibited by whole blood components typical in certain crude sample preparations . Moreover, the thermostability of the enzyme compared to alternative DNA polymerases (Bst and reverse transcriptases allows pretreatment of complete reaction mixes immediately prior to amplification, which facilitates amplification of highly structured genome regions. Compared to Bst, OmniAmp Pol has a faster time to result, particularly with more dilute templates. Molecular diagnostics in field settings can be challenging due to the lack of refrigeration. The stability of OmniAmp Pol is compatible with a dry format that enables long term storage at ambient temperatures. A final requirement for field operability is compatibility with either commonly available instruments or, in other cases, a simple, inexpensive, portable detection mode requiring minimal training or power. Detection of amplification products is shown using lateral flow strips and analysis on a real-time PCR instrument. Results of this study show that OmniAmp Pol is ideally suited for low resource molecular

  10. Novel bioluminescent quantitative detection of nucleic acid amplification in real-time.

    Directory of Open Access Journals (Sweden)

    Olga A Gandelman

    Full Text Available BACKGROUND: The real-time monitoring of polynucleotide amplification is at the core of most molecular assays. This conventionally relies on fluorescent detection of the amplicon produced, requiring complex and costly hardware, often restricting it to specialised laboratories. PRINCIPAL FINDINGS: Here we report the first real-time, closed-tube luminescent reporter system for nucleic acid amplification technologies (NAATs enabling the progress of amplification to be continuously monitored using simple light measuring equipment. The Bioluminescent Assay in Real-Time (BART continuously reports through bioluminescent output the exponential increase of inorganic pyrophosphate (PPi produced during the isothermal amplification of a specific nucleic acid target. BART relies on the coupled conversion of inorganic pyrophosphate (PPi produced stoichiometrically during nucleic acid synthesis to ATP by the enzyme ATP sulfurylase, and can therefore be coupled to a wide range of isothermal NAATs. During nucleic acid amplification, enzymatic conversion of PPi released during DNA synthesis into ATP is continuously monitored through the bioluminescence generated by thermostable firefly luciferase. The assay shows a unique kinetic signature for nucleic acid amplifications with a readily identifiable light output peak, whose timing is proportional to the concentration of original target nucleic acid. This allows qualitative and quantitative analysis of specific targets, and readily differentiates between negative and positive samples. Since quantitation in BART is based on determination of time-to-peak rather than absolute intensity of light emission, complex or highly sensitive light detectors are not required. CONCLUSIONS: The combined chemistries of the BART reporter and amplification require only a constant temperature maintained by a heating block and are shown to be robust in the analysis of clinical samples. Since monitoring the BART reaction requires only a

  11. Multiplexed Recombinase Polymerase Amplification Assay To Detect Intestinal Protozoa.

    Science.gov (United States)

    Crannell, Zachary; Castellanos-Gonzalez, Alejandro; Nair, Gayatri; Mejia, Rojelio; White, A Clinton; Richards-Kortum, Rebecca

    2016-02-01

    This work describes a proof-of-concept multiplex recombinase polymerase amplification (RPA) assay with lateral flow readout that is capable of simultaneously detecting and differentiating DNA from any of the diarrhea-causing protozoa Giardia, Cryptosporidium, and Entamoeba. Together, these parasites contribute significantly to the global burden of diarrheal illness. Differential diagnosis of these parasites is traditionally accomplished via stool microscopy. However, microscopy is insensitive and can miss up to half of all cases. DNA-based diagnostics such as polymerase chain reaction (PCR) are far more sensitive; however, they rely on expensive thermal cycling equipment, limiting their availability to centralized reference laboratories. Isothermal DNA amplification platforms, such as the RPA platform used in this study, alleviate the need for thermal cycling equipment and have the potential to broaden access to more sensitive diagnostics. Until now, multiplex RPA assays have not been developed that are capable of simultaneously detecting and differentiating infections caused by different pathogens. We developed a multiplex RPA assay to detect the presence of DNA from Giardia, Cryptosporidium, and Entamoeba. The multiplex assay was characterized using synthetic DNA, where the limits-of-detection were calculated to be 403, 425, and 368 gene copies per reaction of the synthetic Giardia, Cryptosporidium, and Entamoeba targets, respectively (roughly 1.5 orders of magnitude higher than for the same targets in a singleplex RPA assay). The multiplex assay was also characterized using DNA extracted from live parasites spiked into stool samples where the limits-of-detection were calculated to be 444, 6, and 9 parasites per reaction for Giardia, Cryptosporidium, and Entamoeba parasites, respectively. This proof-of-concept assay may be reconfigured to detect a wide variety of targets by re-designing the primer and probe sequences.

  12. LOMA: A fast method to generate efficient tagged-random primers despite amplification bias of random PCR on pathogens

    Directory of Open Access Journals (Sweden)

    Lee Wah

    2008-09-01

    Full Text Available Abstract Background Pathogen detection using DNA microarrays has the potential to become a fast and comprehensive diagnostics tool. However, since pathogen detection chips currently utilize random primers rather than specific primers for the RT-PCR step, bias inherent in random PCR amplification becomes a serious problem that causes large inaccuracies in hybridization signals. Results In this paper, we study how the efficiency of random PCR amplification affects hybridization signals. We describe a model that predicts the amplification efficiency of a given random primer on a target viral genome. The prediction allows us to filter false-negative probes of the genome that lie in regions of poor random PCR amplification and improves the accuracy of pathogen detection. Subsequently, we propose LOMA, an algorithm to generate random primers that have good amplification efficiency. Wet-lab validation showed that the generated random primers improve the amplification efficiency significantly. Conclusion The blind use of a random primer with attached universal tag (random-tagged primer in a PCR reaction on a pathogen sample may not lead to a successful amplification. Thus, the design of random-tagged primers is an important consideration when performing PCR.

  13. Marketability of ready-to-eat cactus pear as affected by temperature and modified atmosphere.

    Science.gov (United States)

    Cefola, Maria; Renna, Massimiliano; Pace, Bernardo

    2014-01-01

    In order to increase the diffusion of cactus pear fruits, in this study, the proper maturity index for peeling and processing them as ready-to-eat product was evaluated and characterized. Thereafter, the effects of different storage temperatures and modified atmosphere conditions on the marketability of ready-to-eat cactus pear were studied. The storage of ready-to-eat fruits at 4 °C in both passive (air) and semi-active (10 kPa O2 and 10 kPa CO2) modified atmosphere improved the marketability by 30%, whereas the storage at 8 °C caused a dangerous reduction in O2 partial pressure inside modified atmosphere packages, due to fruits' increased metabolic activity. A very low level of initial microbial growth was detected, while a severe increase in mesophilic and psychrophilic bacteria was shown in control samples at both temperatures during storage; an inhibitory effect of modified atmosphere on microbial growth was also observed. In conclusion, modified atmosphere improved only the marketability of fruits stored at 4 °C; whereas the storage at 8 °C resulted in deleterious effects on the ready-to-eat fruits, whether stored in air or in modified atmosphere.

  14. Quality Analysis of Pear Fruit of Shah Miveh variety Using Nondestructive Ultrasonic Technique

    Directory of Open Access Journals (Sweden)

    R Meamar Dastjerdi

    2014-09-01

    Full Text Available Development of ultrasound technique has not been progressing for evaluating the internal quality of fruits as fast as that of processed foods. In this research for quality assessment of pear fruit (Shah Miveh variety an ultrasonic measurement system was constructed to transmit and receive the ultrasonic waves. The apparatus included a pulser-receiver, a pair of 75 kHz ultrasonic transducers with exponential horn, and a computer system for data acquisition and analysis. Several mechanical and chemical properties, including firmness, TSS, acidity, elastic modulus, pH and total dry matter for destructive quality assessment were measured. Velocity and attenuation of ultrasonic waves for nondestructive tests were also measured. The fruit quality levels for the experiment were: unripe, ripe and overripe. The results of tests showed that firmness was the best parameter for measuring fruit quality, as it decreased significantly with ripeness. The effect of ripeness on the velocity and attenuation of ultrasonic waves was also significant. Investigation showed a positive linear relationship between fruit firmness and wave velocity (R2=0.81. Furthermore, the relationship between fruit firmness and attenuation was exponential and wave attenuation decreased with increasing fruit firmness (R2=0.895. The Relationship between ultrasonic properties and fruit modulus of elasticity showed that the wave velocity increased and attenuation decreased with ‌increasing elasticity. It can be concluded that the ultrasonic instrument equipped with exponential horns can effectively be utilized for pear quality assessment based on measurement of wave velocity and attenuation.

  15. Feeding behavior and performance of sheep fed cactus pear in substitution of corn

    Directory of Open Access Journals (Sweden)

    Roberto Germano Costa

    2013-11-01

    Full Text Available The objective of this study was to evaluate the feeding behavior and performance of Santa Ines sheep subjected to different levels of substitution of corn by cactus pear in the diet. Forty-fivenon-castrated male Santa Inês sheep with initial live weight of 27.50±0.48 kg were distributed in a completely randomized design with five treatments (0, 70, 140, 210 and 280 g/kg DM and nine replicates. Dry matter and neutral detergent fiberintakes showed quadratic behavior. Times spent eating, ruminating and total ruminating chews showed increasing linear behavior, while the idle time decreased with increasing amounts of dietary cactus. The feeding efficiency (gDM/h increased linearly, while differences in rumination efficiency of the DM (g DM/h; NDF intake efficiency (gNDF/h and NDF rumination efficiency (gNDF/h were not significant. There was no significant effect for the number of ruminated boli and number of ruminating chews per bolus. The number of chews per day increased linearly. These results indicate that cactus pear in substitution of corn had no influence on the feeding behavior of feedlot sheep.

  16. Performance of orange oil in the control of carmine cochineal in giant cactus pear.

    Directory of Open Access Journals (Sweden)

    Ivanildo Cavalcanti de Albuquerque

    2009-03-01

    Full Text Available Since its introduction, in 2001, the carmine cochineal (Dactylopius opuntiae already decimated some 100.000 hectares of giant cactus pear (Opuntia ficus-indica in semi-arid region of Paraiba. This study aimed to evaluate the behavior of five concentrations of orange oil, applied in cladodes on the death of D. opuntiae in field conditions. The research was carried out in a field of giant cactus pear infested by carmine cochineal on the site rigideira, Monteiro County, State of Paraíba. The trial design used was blocks at random (DBR composed of six treatments [doses of 0.3, 0.4, 0.5, 0.6, and 0.7% of orange oil (Prev-am] and water as control and five repetitions. The orange oil known like Prev-Am (Sodium tetraborohydrate decahydrate was effective against to carmine cochineal as early as the dose of 0.3% and higher potential for efficiency were observed at doses of 0.6 and 0.7%. After 48 hours of application of the product, which was observed at doses applied adults and nymphs of the insect, was dried according to the product action that acts by contact. The product had no lethal effect on ladybugs (Cycloneda sanguinea and Scymnus intrusus, but was lethal to larvae of Baccha sp. at a dose of 0.7%.

  17. Dispersal speed of datylopius opuntiae on giant cactus pear (opuntia fícus- indica

    Directory of Open Access Journals (Sweden)

    Carlos Henrique de Brito

    2009-08-01

    Full Text Available The insect Dactylopius opuntiae (cochineal carmine has become an important pest to giant cactus pear (Opuntia ficus-indica in several counties of the micro regions of Carirí Ocidental, Serra do Teixeira and Piancó, where the attack of the insect is so intense that it obliges farmers to eradicate crops. This research aimed to quantify the dispersal speed of D. opuntiae under field conditions, as a premise for the implementation of tactics of the Integrated Pest Management (IPM. The experiment was carried out at the Lagoa Seca Experimental Station, in Lagoa Seca County, state of Paraiba. Dispersion quantification was conducted in three rows of giant cactus pear each with ten plants, the first being selected to perform the artificial infestation (initial. Three evaluations was carried out in three rows and counted the average number of colonies arising from the initial infestation. Medium comparison of was made by Tukey test at 5% probability, using the application ASSISTAT 7.5 Beta. For the aspect of dispersion within each plant, it was observed that the artificially infested cladodes began to be colonized for 8 days after infection and subsequently at 15, 21, 28, 35 and 42 and 50 days, noting that equally the first, second and third rows were also colonized, showing thus the dispersal speed of the insect pest.

  18. Erwinia tasmaniensis sp. nov., a non-phytopathogenic bacterium from apple and pear trees.

    Science.gov (United States)

    Geider, Klaus; Auling, Georg; Du, Zhiqiang; Jakovljevic, Vladimir; Jock, Susanne; Völksch, Beate

    2006-12-01

    Bacteria were isolated from flowers and bark of apple and pear trees at three places in Australia. In Victoria, Tasmania and Queensland, strains with white colonies on nutrient agar were screened for dome-shaped colony morphology on agar with sucrose and were found to be closely related by several criteria. The isolates were not pathogenic on apples or pears. They were characterized by a polyphasic approach including microbiological and API assays as well as fatty acid methyl ester analysis, DNA-DNA hybridization and DNA sequencing. For molecular classification, the 16S rRNA cistron and the conserved genes gpd and recA of these bacteria were investigated. Together with other taxonomic criteria, the results of these studies indicate that the bacteria belong to a novel separate species, which we propose to name Erwinia tasmaniensis sp. nov., with the type strain Et1/99(T) (=DSM 17950(T)=NCPPB 4357(T)). From DNA-DNA hybridization kinetics, microbiological characteristics and nucleotide sequence analyses, this species is related to pathogenic Erwinia species, but also to the epiphytic species Erwinia billingiae.

  19. Effect of planting methods on cladodes production in sweet cactus pear.

    Directory of Open Access Journals (Sweden)

    Ivanildo Cavalcatni de Albuquerque

    2009-03-01

    Full Text Available In the Northeast of Brazil, are grown predominantly two species of cactus pear, the Nopalea cochenillifera and Opuntia ficus indica. In the last two decades, the growing interest in and knowledge of fodder have greatly increased by the farmers. The objective of this research was to investigate how best method to plant the sweet cactus pear, which produces more cladodes per plant, from the mother cladodes. The experiment was conducted in Lagoa Seca-PB county, at field level, at Lagoa Seca Experimental Station from EMEPA-PB. The genotype used was Palmepa - PB1 (Baiana planted at a spacing of 1 x 50 m and in a soil classified as Neossol Regolithic Eutrophic. The cladodes were planted according to three types of plantation: P1 - cladodes planted upright 90°, P2 - cladodes planted with apex to the east, inclination of 45º and P3 - cladodes planted with apex to the west, with inclination of 45º. It was found that there was no statistical difference between treatments, but the planting method with the cladodes planted vertically (P1 showed, in 300 plants, an average of 134 and 109 producing, more cladodes in relation to planting method P2 and P3, respectively.

  20. Nutrition and yield of ‘Gigante’ cactus pear cultivated with different spacings and organic fertilizer

    Directory of Open Access Journals (Sweden)

    Paulo E. R. Donato

    Full Text Available ABSTRACT The objective of this study was to evaluate the levels of macronutrients in cladodes and yield of cactus pear, cv. ‘Gigante’, cultivated with different cattle manure doses and plant spacings. The experimental design was randomized blocks in 4 x 3 factorial, with three replicates. The treatments consisted of the combination of four doses of cattle manure (0, 30, 60 and 90 Mg ha-1 year-1 with three spacings (1.00 x 0.50, 2.00 x 0.25 and 3.00 x 1.00 x 0.25 m. The contents of macronutrients and dry matter production of cladodes were assessed 600 days after planting. The plant spacings influenced the contents of nitrogen, potassium, calcium and sulfur in the cladodes of ‘Gigante’ cactus pear and there was interaction between spacing and manure dose for magnesium contents. The increment in cattle manure doses increases the contents of phosphorus, nitrogen, potassium and sulfur in the cladodes. The maximum dry matter production of cladodes is estimated at 21.8 Mg ha-1 year-1 at a dose of 71.8 Mg ha-1 year-1 of manure.

  1. Electricity-free amplification and detection for molecular point-of-care diagnosis of HIV-1.

    Science.gov (United States)

    Singleton, Jered; Osborn, Jennifer L; Lillis, Lorraine; Hawkins, Kenneth; Guelig, Dylan; Price, Will; Johns, Rachel; Ebels, Kelly; Boyle, David; Weigl, Bernhard; LaBarre, Paul

    2014-01-01

    In resource-limited settings, the lack of decentralized molecular diagnostic testing and sparse access to centralized medical facilities can present a critical barrier to timely diagnosis, treatment, and subsequent control and elimination of infectious diseases. Isothermal nucleic acid amplification methods, including reverse transcription loop-mediated isothermal amplification (RT-LAMP), are well-suited for decentralized point-of-care molecular testing in minimal infrastructure laboratories since they significantly reduce the complexity of equipment and power requirements. Despite reduced complexity, however, there is still a need for a constant heat source to enable isothermal nucleic acid amplification. This requirement poses significant challenges for laboratories in developing countries where electricity is often unreliable or unavailable. To address this need, we previously developed a low-cost, electricity-free heater using an exothermic reaction thermally coupled with a phase change material. This heater achieved acceptable performance, but exhibited considerable variability. Furthermore, as an enabling technology, the heater was an incomplete diagnostic solution. Here we describe a more precise, affordable, and robust heater design with thermal standard deviation electricity-free heater and NALF-detection platform, we demonstrate sensitive and repeatable detection of HIV-1 with a ß-actin positive internal amplification control from processed sample to result in less than 80 minutes. Together, these elements are building blocks for an electricity-free platform capable of isothermal amplification and detection of a variety of pathogens.

  2. 生态健康果园(香梨)中梨小食心虫发生动态监测研究%Dynamic monitoring of pear moth in ecological health orchard (fragrant pear)

    Institute of Scientific and Technical Information of China (English)

    李健; 张磊; 彭永杰; 毕司进

    2014-01-01

    The occurrence principle of pear moth in ecological health orchard were monitored with sex pheromone and sweet and sour liquid in order to provide the reference for the effective monitoring and measures establishment of biological prevention and control in health orchard (fragrant pear) and conventional orchard. Results show that the law trends of growth and decline of pear moth for the two kinds of traps are same. Pear moth occurs 4 generations a year with obvious peak. The emergence peak of overwintering adults is in the middle and last ten-day of a month. Because of the existence of sex pheromone in the ecological health orchard (fragrant pear), pear moth adult is more sensitive to sweet and sour liquid than sex pheromone. The sweet and sour liquid trap is good for monitoring the dynamic happening of the pear moth, and the accuracy is significantly higher than the sex pheromone trap.%通过对生态健康果园(香梨)中利用性信息素和糖醋液诱捕器对梨小食心虫的发生动态进行监测研究,为生态健康果园(香梨)及常规果园的有效监测和适时制定生物防治措施提供参考。研究表明:两种诱捕器对梨小食心虫全年发生消长规律监测的趋势基本一致,梨小食心虫1年发生4代,峰值明显,越冬代羽化高峰期为4月中、下旬。在生态健康果园(香梨)中由于大量、高浓度性信息素的存在,梨小食心虫成虫对糖醋液的趋性比性信息素的趋性敏感,利用糖醋液诱捕器能够很好地监测梨小食心虫的发生动态,且监测准确性明显高于性信息素诱捕器。

  3. 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.

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

    Science.gov (United States)

    Cao, Yunpeng; Han, Yahui; Li, Dahui; Lin, Yi; Cai, Yongping

    2016-10-19

    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.

  5. Time varying arctic climate change amplification

    Energy Technology Data Exchange (ETDEWEB)

    Chylek, Petr [Los Alamos National Laboratory; Dubey, Manvendra K [Los Alamos National Laboratory; Lesins, Glen [DALLHOUSIE U; Wang, Muyin [NOAA/JISAO

    2009-01-01

    During the past 130 years the global mean surface air temperature has risen by about 0.75 K. Due to feedbacks -- including the snow/ice albedo feedback -- the warming in the Arctic is expected to proceed at a faster rate than the global average. Climate model simulations suggest that this Arctic amplification produces warming that is two to three times larger than the global mean. Understanding the Arctic amplification is essential for projections of future Arctic climate including sea ice extent and melting of the Greenland ice sheet. We use the temperature records from the Arctic stations to show that (a) the Arctic amplification is larger at latitudes above 700 N compared to those within 64-70oN belt, and that, surprisingly; (b) the ratio of the Arctic to global rate of temperature change is not constant but varies on the decadal timescale. This time dependence will affect future projections of climate changes in the Arctic.

  6. Amplification, Redundancy, and Quantum Chernoff Information

    Science.gov (United States)

    Zwolak, Michael; Riedel, C. Jess; Zurek, Wojciech H.

    2014-04-01

    Amplification was regarded, since the early days of quantum theory, as a mysterious ingredient that endows quantum microstates with macroscopic consequences, key to the "collapse of the wave packet," and a way to avoid embarrassing problems exemplified by Schrödinger's cat. Such a bridge between the quantum microworld and the classical world of our experience was postulated ad hoc in the Copenhagen interpretation. Quantum Darwinism views amplification as replication, in many copies, of the information about quantum states. We show that such amplification is a natural consequence of a broad class of models of decoherence, including the photon environment we use to obtain most of our information. This leads to objective reality via the presence of robust and widely accessible records of selected quantum states. The resulting redundancy (the number of copies deposited in the environment) follows from the quantum Chernoff information that quantifies the information transmitted by a typical elementary subsystem of the environment.

  7. On Arbitrary Phases in Quantum Amplitude Amplification

    CERN Document Server

    Hoyer, P

    2000-01-01

    We consider the use of arbitrary phases in quantum amplitude amplification which is a generalization of quantum searching. We prove that the phase condition in amplitude amplification is given by $\\tan(\\phi/2)=\\tan(\\phi/2)(1-2a)$, where $\\phi$ and $\\phi$ are the phases used and where $a$ is the success probability of the given algorithm. Thus the choice of phases depends nontrivially and nonlinearly on the success probability. Utilizing this condition, we give methods for constructing quantum algorithms that succeed with certainty and for implementing arbitrary rotations. We also conclude that phase errors of order up to $\\frac{1}{\\sqrt{a}}$ can be tolerated in amplitude amplification.

  8. An integrated closed-tube 2-plex PCR amplification and hybridization assay with switchable lanthanide luminescence based spatial detection.

    Science.gov (United States)

    Lahdenperä, Susanne; Spangar, Anni; Lempainen, Anna-Maija; Joki, Laura; Soukka, Tero

    2015-06-21

    Switchable lanthanide luminescence is a binary probe technology that inherently enables a high signal modulation in separation-free detection of DNA targets. A luminescent lanthanide complex is formed only when the two probes hybridize adjacently to their target DNA. We have now further adapted this technology for the first time in the integration of a 2-plex polymerase chain reaction (PCR) amplification and hybridization-based solid-phase detection of the amplification products of the Staphylococcus aureus gyrB gene and an internal amplification control (IAC). The assay was performed in a sealed polypropylene PCR chip containing a flat-bottom reaction chamber with two immobilized capture probe spots. The surface of the reaction chamber was functionalized with NHS-PEG-azide and alkyne-modified capture probes for each amplicon, labeled with a light harvesting antenna ligand, and covalently attached as spots to the azide-modified reaction chamber using a copper(i)-catalyzed azide-alkyne cycloaddition. Asymmetric duplex-PCR was then performed with no template, one template or both templates present and with a europium ion carrier chelate labeled probe for each amplicon in the reaction. After amplification europium fluorescence was measured by scanning the reaction chamber as a 10 × 10 raster with 0.6 mm resolution in time-resolved mode. With this assay we were able to co-amplify and detect the amplification products of the gyrB target from 100, 1000 and 10,000 copies of isolated S. aureus DNA together with the amplification products from the initial 5000 copies of the synthetic IAC template in the same sealed reaction chamber. The addition of 10,000 copies of isolated non-target Escherichia coli DNA in the same reaction with 5000 copies of the synthetic IAC template did not interfere with the amplification or detection of the IAC. The dynamic range of the assay for the synthetic S. aureus gyrB target was three orders of magnitude and the limit of detection of 8 p

  9. Potential of osmoadaptation for improving Pantoea agglomerans E325 as biocontrol agent for fire blight of apple and pear

    Science.gov (United States)

    Pantoea agglomerans biocontrol strain E325 is the active ingredient in a commercial product for fire blight, a destructive disease of apple and pear initiated by Erwinia amylovora in flowers. Osmoadaptation, involving the combination of saline osmotic stress and osmolyte amendment to growth media, w...

  10. Extraction of arbutin and its comparative content in branches, leaves, stems, and fruits of Japanese pear Pyrus pyrifolia cv. Kousui.

    Science.gov (United States)

    Sasaki, Chizuru; Ichitani, Masaki; Kunimoto, Ko-Ki; Asada, Chikako; Nakamura, Yoshitoshi

    2014-01-01

    Arbutin is a tyrosinase inhibitor and is extensively used as a human skin-whitening agent. This study investigated the optimum conditions for extracting arbutin by ultrasonic homogenization from discarded branches pruned from Japanese pear (Pyrus pyrifolia cv. Kousui) trees. The arbutin content was measured in the branches and also in the leaves, stems, fruit peel, and fruit flesh.

  11. 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.

  12. Production technology of thorn pear health vinegar%刺梨保健醋生产工艺研究

    Institute of Scientific and Technical Information of China (English)

    杨胜敖

    2009-01-01

    For the development and utilization of rich resources of wild thorn pear, thorn pear juice was used as raw material to research on the production technique of thorn pear vinegar. Through the orthogonal test, the optimal conditions were as follow: sugar 14%, yeast mud 8% and fermentation temperature 32℃ in alcoholic fermentation could get the highest alcohol content; inoculum 10%, alcohol content 7%, fermentation temperature 35℃, in acetic fermentation may make thorn pear vinegar with the characteristic of the sour and sweet tasty, the fruit aroma and the health function.%为开发利用丰富的野生刺梨资源,以刺梨汁为原料,对刺梨醋的生产工艺进行了研究.通过正交试验优选最佳生产工艺条件,结果表明:酒精发酵的糖度14%、酵母液用量8%、发酵温度32℃,产生的酒精量最多;醋酸发酵的接种量10%、酒精浓度6%、发酵温度35℃,可制得酸甜爽口、果香突出、营养丰富和独具保健功能的刺梨醋.

  13. 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 (

  14. Epidemiology and effective control of Altenaria altenata, causal agent of dead (dormant) flower bud disease of pear

    NARCIS (Netherlands)

    Wenneker, M.; Joosten, N.N.; Anbergen, R.H.N.; Vink, P.; Bruggen, van A.S.

    2011-01-01

    Dead flower buds are a common phenomenon in pear culture in The Netherlands, Belgium and Mediterranean countries. Disease cases are also reported from South America. The disease is characterized by a partial or complete necrosis of flower buds during tree dormancy. The disease progresses during wint

  15. Differences in leaf litter, ascospore production and infection of pear scab (Venturia pirina) in Dutch organic orchards

    NARCIS (Netherlands)

    Timmermans, B.G.H.; Jansonius, P.J.

    2012-01-01

    In 2010 and 2011 the amounts of leaf litter and ascospore production per unit of leaf litter area in 7 organic pear orchards throughout the Netherlands were measured. In one of the orchards, adapted managements strategies were implemented, being grass/clover that is grown as ground cover on the tree

  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. [Characteristic wavelengths selection of soluble solids content of pear based on NIR spectral and LS-SVM].

    Science.gov (United States)

    Fan, Shu-xiang; Huang, Wen-qian; Li, Jiang-bo; Zhao, Chun-jiang; Zhang, Bao-hua

    2014-08-01

    To improve the precision and robustness of the NIR model of the soluble solid content (SSC) on pear. The total number of 160 pears was for the calibration (n=120) and prediction (n=40). Different spectral pretreatment methods, including standard normal variate (SNV) and multiplicative scatter correction (MSC) were used before further analysis. A combination of genetic algorithm (GA) and successive projections algorithm (SPA) was proposed to select most effective wavelengths after uninformative variable elimination (UVE) from original spectra, SNV pretreated spectra and MSC pretreated spectra respectively. The selected variables were used as the inputs of least squares-support vector machine (LS-SVM) model to build models for de- termining the SSC of pear. The results indicated that LS-SVM model built using SNVE-UVE-GA-SPA on 30 characteristic wavelengths selected from full-spectrum which had 3112 wavelengths achieved the optimal performance. The correlation coefficient (Rp) and root mean square error of prediction (RMSEP) for prediction sets were 0.956, 0.271 for SSC. The model is reliable and the predicted result is effective. The method can meet the requirement of quick measuring SSC of pear and might be important for the development of portable instruments and online monitoring.

  18. Mesos components (CaC12, MgSO4, KH2P04) are critical for improving pear micropropagation

    Science.gov (United States)

    The USDA-ARS National Clonal Germplasm Repository in vitro collection contains over 200 pear accessions in 18 species. Due to the wide genetic diversity of this collection there is also a diverse response to growth on standard tissue culture media. An initial study of mineral nutrition using a syste...

  19. 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...

  20. 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

  1. 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.

  2. UV-C light inactivation kinetics of Penicillium expansum on pear surfaces: Influence on physicochemical and sensory quality during storage

    Science.gov (United States)

    Postharvest quality and storage life of fresh pear are often limited by fungal growth caused by Penicillium expansum. Ultraviolet-C light (UV-C 254 nm) is a promising alternative disinfestation method to reduce fruit spoilage by fungi. In this study, UV-C inactivation kinetic data of Penicillium exp...

  3. Purification and biochemical characterization of polygalacturonase produced by Penicillium expansum during postharvest decay of ‘Anjou’ pear

    Science.gov (United States)

    A polygalacturonase (PG) was extracted and purified from decayed tissue of ‘Anjou’ pear fruit inoculated with Penicillium expansum. Ammonium sulfate precipitation, gel filtration and cation exchange chromatography were used to purify the enzyme. Both chromatographic methods revealed a single peak co...

  4. Current management efforts against Cactoblastis cactorum as a pest of North American prickly pear cactus, Opuntia spp.

    Science.gov (United States)

    The unintentional arrival of Cactoblastis cactorum (Lepidoptera: Pyralidae) to Florida changed the scope of this celebrated weed biological control agent from savior to pest. Based on this insects’ substantial control of non-native Opuntia spp. (prickly pear cactus) in Australia and other parts of ...

  5. 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

  6. Radiopolymerization of {beta}(-)pinene: A case of chiral amplification

    Energy Technology Data Exchange (ETDEWEB)

    Cataldo, Franco [Soc. Lupi Chemical Research, Via Casilina 1626/A, 00133 Rome (Italy)]. E-mail: cdcata@flashnet.it; Keheyan, Yeghis [CNR, Istituto per lo studio dei Materiali Nanostrutturati, Department of Chemistry, University ' La Sapienza' , P.le Aldo Moro 1, Rome (Italy)

    2006-05-15

    {beta}(-)Pinene was treated with {gamma} radiation at three dose levels: 150, 300 and 600 kGy. The expected effect of radiation at these high doses was the partial racemization of the substrate as already observed in the case of other terpene monomers. Unexpectedly {beta}(-)pinene underwent a radiopolymerization reaction into a solid resin and into a dimer. The structure of the products was studied by FT-IR spectroscopy also in comparison to a reference {beta}(-)pinene resin prepared by cationic polymerization. A highly ordered structure was found in the case of the radiopolymer in comparison to the resin from cationic polymerization. Polarimetric measurements have shown astonishing enhancement in the optical activity of the radiopolymer and radiodimer in comparison to the starting optical activity of the {beta}(-)pinene monomer. The results have been discussed in terms of amplification of chirality caused by {gamma} radiation and the implications of this fact on the mechanism of chiral amplification on prebiotic molecules.

  7. Continuous phase amplification with a Sagnac interferometer

    CERN Document Server

    Starling, David J; Williams, Nathan S; Jordan, Andrew N; Howell, John C

    2009-01-01

    We describe a weak value inspired phase amplification technique in a Sagnac interferometer. We monitor the relative phase between two paths of a slightly misaligned interferometer by measuring the average position of a split-Gaussian mode in the dark port. Although we monitor only the dark port, we show that the signal varies linearly with phase and that we can obtain similar sensitivity to balanced homodyne detection. We derive the source of the amplification both with classical wave optics and as an inverse weak value.

  8. Effective Privacy Amplification for Secure Classical Communications

    CERN Document Server

    Horvath, Tamas; Scheuer, Jacob

    2011-01-01

    We study the effectiveness of privacy amplification for classical key-distribution schemes. We find that, unlike quantum key distribution schemes, the high fidelity of the raw key in classical systems allow the users to always sift a secure shorter key, given that they have an upper bound of eavesdropper probability to correctly guess the exchanged key-bits. We establish the number of privacy amplification iterations needed to achieve information leak of 10^-8 in several classical systems and highlight the inherent tradeoff between the number of iterations and the security of the raw key.

  9. Parametric Amplification For Detecting Weak Optical Signals

    Science.gov (United States)

    Hemmati, Hamid; Chen, Chien; Chakravarthi, Prakash

    1996-01-01

    Optical-communication receivers of proposed type implement high-sensitivity scheme of optical parametric amplification followed by direct detection for reception of extremely weak signals. Incorporates both optical parametric amplification and direct detection into optimized design enhancing effective signal-to-noise ratios during reception in photon-starved (photon-counting) regime. Eliminates need for complexity of heterodyne detection scheme and partly overcomes limitations imposed on older direct-detection schemes by noise generated in receivers and by limits on quantum efficiencies of photodetectors.

  10. Effect of pyrimethanil on Cryptococcus laurentii, Rhodosporidium paludigenum, and Rhodotorula glutinis biocontrol of Penicillium expansum infection in pear fruit.

    Science.gov (United States)

    Yu, Chen; Zhou, Tao; Sheng, Kuang; Zeng, Lizhen; Ye, Changzhou; Yu, Ting; Zheng, Xiaodong

    2013-06-17

    The effect of biocontrol yeasts and pyrimethanil at low concentration on inhibition of blue mold rot caused by Penicillium expansum in pear fruit was investigated. Pyrimethanil at low concentration (40μg/mL) alone had little inhibitory activity against the P. expansum infection in pear fruit wounds although it was effective in inhibiting the survival of P. expansum on Asp-agar medium. Pyrimethanil at this low concentration significantly enhanced the efficacy of Cryptococcus laurentii at 1×10(7)CFU/mL in reducing blue mold rot in vivo compared with C. laurentii at 1×10(7)CFU/mL alone. However, there was no additive inhibitory activity when pyrimethanil was combined for application with biocontrol yeasts Rhodosporidium paludigenum or Rhodotorula glutinis. Combination of pyrimethanil and C. laurentii at low concentration also inhibited blue mold rot when P. expansum was inoculated into fruit wounds 12h before treatment and fruit was stored at low temperature (4°C). Pyrimethanil at 0.04 to 400μg/mL did not influence the survival of C. laurentii in vitro, and it only slightly reduced the population growth of C. laurentii after 48h of incubation in the pear fruit wounds. There was no significant difference in quality parameters including total soluble solids, titratable acidity and ascorbic acid of pear fruit wounds among all treatments after 5days of treatment at 25°C. Integration of C. laurentii and pyrimethanil at low concentration might be an effective and safe strategy to control P. expansum infection in pear fruit, especially in an integrated postharvest disease management strategy.

  11. Detection of Entamoeba histolytica by Recombinase Polymerase Amplification

    Science.gov (United States)

    Nair, Gayatri; Rebolledo, Mauricio; White, A. Clinton; Crannell, Zachary; Richards-Kortum, R. Rebecca; Pinilla, A. Elizabeth; Ramírez, Juan David; López, M. Consuelo; Castellanos-Gonzalez, Alejandro

    2015-01-01

    Amebiasis is an important cause of diarrheal disease worldwide and has been associated with childhood malnutrition. Traditional microscopy approaches are neither sensitive nor specific for Entamoeba histolytica. Antigen assays are more specific, but many cases are missed unless tested by molecular methods. Although polymerase chain reaction (PCR) is effective, the need for sophisticated, expensive equipment, infrastructure, and trained personnel limits its usefulness, especially in the resource-limited, endemic areas. Here, we report development of a recombinase polymerase amplification (RPA) method to detect E. histolytica specifically. Using visual detection by lateral flow (LF), the test was highly sensitive and specific and could be performed without additional equipment. The availability of this inexpensive, sensitive, and field-applicable diagnostic test could facilitate rapid diagnosis and treatment of amebiasis in endemic regions. PMID:26123960

  12. Detection of Entamoeba histolytica by Recombinase Polymerase Amplification.

    Science.gov (United States)

    Nair, Gayatri; Rebolledo, Mauricio; White, A Clinton; Crannell, Zachary; Richards-Kortum, R Rebecca; Pinilla, A Elizabeth; Ramírez, Juan David; López, M Consuelo; Castellanos-Gonzalez, Alejandro

    2015-09-01

    Amebiasis is an important cause of diarrheal disease worldwide and has been associated with childhood malnutrition. Traditional microscopy approaches are neither sensitive nor specific for Entamoeba histolytica. Antigen assays are more specific, but many cases are missed unless tested by molecular methods. Although polymerase chain reaction (PCR) is effective, the need for sophisticated, expensive equipment, infrastructure, and trained personnel limits its usefulness, especially in the resource-limited, endemic areas. Here, we report development of a recombinase polymerase amplification (RPA) method to detect E. histolytica specifically. Using visual detection by lateral flow (LF), the test was highly sensitive and specific and could be performed without additional equipment. The availability of this inexpensive, sensitive, and field-applicable diagnostic test could facilitate rapid diagnosis and treatment of amebiasis in endemic regions.

  13. Simplified procedures for applying the polymerase chain reaction to routinely fixed paraffin wax sections.

    Science.gov (United States)

    Coates, P J; d'Ardenne, A J; Khan, G; Kangro, H O; Slavin, G

    1991-02-01

    The polymerase chain reaction was applied to the analysis of DNA contained in archival paraffin wax embedded material. DNA suitable for the reaction was obtained from these tissues by simple extraction methods, without previous dewaxing of tissue sections. When compared with unfixed material, the reaction efficiency was compromised, so that an increased number of amplification cycles were required to produce equivalent amounts of amplified product. This in turn led to an increase in amplification artefacts, which can be minimised by a simple modification of the standard reaction. Amplification of relatively large DNA fragments was not always successful, and it seems prudent to bear this in mind when designing oligonucleotide primers which are to be used for the amplification of archival material. The efficiency of the procedure can be improved by dividing the amplification cycles into two parts: this reduces the amount of reagent needed, is relatively simple and inexpensive, and can be performed in one working day.

  14. Intelligence amplification framework for enhancing scheduling processes

    NARCIS (Netherlands)

    Dobrkovic, Andrej; Liu, Luyao; Iacob, Maria-Eugenia; Hillegersberg, van 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 partn

  15. Social amplification of risk: a conceptual framework

    Energy Technology Data Exchange (ETDEWEB)

    Kasperson, R.E.; Renn, O.; Slovic, P.; Brown, H.S.; Emel, J.; Goble, R.; Kasperson, J.X.; Ratick, S.

    1988-06-01

    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. Desert Amplification in a Warming Climate

    Science.gov (United States)

    Zhou, Liming

    2016-08-01

    Here I analyze the observed and projected surface temperature anomalies over land between 50°S-50°N for the period 1950–2099 by large-scale ecoregion and find strongest warming consistently and persistently seen over driest ecoregions such as the Sahara desert and the Arabian Peninsula during various 30-year periods, pointing to desert amplification in a warming climate. This amplification enhances linearly with the global mean greenhouse gases(GHGs) radiative forcing and is attributable primarily to a stronger GHGs-enhanced downward longwave radiation forcing reaching the surface over drier ecoregions as a consequence of a warmer and thus moister atmosphere in response to increasing GHGs. These results indicate that desert amplification may represent a fundamental pattern of global warming associated with water vapor feedbacks over land in low- and mid- latitudes where surface warming rates depend inversely on ecosystem dryness. It is likely that desert amplification might involve two types of water vapor feedbacks that maximize respectively in the tropical upper troposphere and near the surface over deserts, with both being very dry and thus extremely sensitive to changes of water vapor.

  17. 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

  18. Polimorfismo enzimático nos tecidos de pereira Isoenzymatic variability in pear tissues for fingerprinting

    Directory of Open Access Journals (Sweden)

    JOSÉ CARLOS FACHINELLO

    2000-07-01

    Full Text Available O trabalho foi realizado com o objetivo de avaliar o polimorfismo enzimático em diferentes tecidos de oito cultivares de pereira Pyrus communis L. Os genótipos utilizados fazem parte da coleção de plantas disponíveis na Universidade de Estudos de Bolonha. Para as análises isoenzimáticas foram utilizadas gemas floríferas dormentes no inverno, casca de ramos de um ano obtida de plantas em pleno desenvolvimento, folhas obtidas no início da primavera e folhas de plantas mantidas in vitro. A corrida eletroforética foi realizada em gel de poliacrilamida a gradiente com (5% a 12,5%. Os resultados obtidos com os genótipos utilizados indicaram que o sistema enzimático beta-glucosidase (E.C.3.2.1.21 apresentou atividade apenas nas folhas das plantas in vitro, com uma banda na mesma posição para todas as cultivares, ao passo que os sistemas para as enzimas esterase (E.C.3.1.1.2 e peroxidase (E.C.1.11.1.7 apresentaram elevado polimorfismo. Nas gemas dormentes analisadas, o sistema peroxidase permitiu diferenciar todos os genótipos. As formas isoenzimáticas da esterase permitiram separar todos os genótipos independentemente dos tecidos utilizados.Eight self-rooted genotype of pear (Pyrus communis L. were assessed to assert the patterns of the isoenzymatic systems beta-glucosidase (E.C.3.2.1.21, esterase (E.C.3.1.1.2 and peroxidase (E.C.1.11.1.7. Tissues samples of buds, leaves, shoot bark and in vitro leaves were analyzed by polyacrylamide gel electrophoresis (5% to 12.5%. High variability of esterase isoenzyme was detected among the tissues of each cultivar. The beta-glucosidase activity was detected only in in vitro leaves, otherwise polymorphism was not found. High variability of peroxidase in all pear genotypes was found with isoenzyme extract from buds. The other tissues showed enough variability, although less variability was detected for differentiation of all assayed cultivars. Esterase proved to be most reliable enzyme for

  19. 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.

  20. A Novel Ultrasensitive ECL Sensor for DNA Detection Based on Nicking Endonuclease-Assisted Target Recycling Amplification, Rolling Circle Amplification and Hemin/G-Quadruplex

    Directory of Open Access Journals (Sweden)

    Fukang Luo

    2015-01-01

    Full Text Available In this study, we describe a novel universal and highly sensitive strategy for the electrochemiluminescent (ECL detection of sequence specific DNA at the aM level based on Nt.BbvCI (a nicking endonuclease-assisted target recycling amplification (TRA, rolling circle amplification (RCA and hemin/G-quadruplex. The target DNAs can hybridize with self-assembled capture probes and assistant probes to form “Y” junction structures on the electrode surface, thus triggering the execution of a TRA reaction with the aid of Nt.BbvCI. Then, the RCA reaction and the addition of hemin result in the production of numerous hemin/G-quadruplex, which consume the dissolved oxygen in the detection buffer and result in a significant ECL quenching effect toward the O2/S2O82− system. The proposed strategy combines the amplification ability of TRA, RCA and the inherent high sensitivity of the ECL technique, thus enabling low aM (3.8 aM detection for sequence-specific DNA and a wide linear range from 10.0 aM to 1.0 pM. At the same time, this novel strategy shows high selectivity against single-base mismatch sequences, which makes our novel universal and highly sensitive method a powerful addition to specific DNA sequence detection.

  1. A new evolutionary theory deduced mathematically from entropy amplification

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    A new evolutionary theory which is able to unite the present evolutionary debates is deduced mathematically from the principle of entropy amplification.It suggests that the extensive evolution is driven by the amplification of entropy,or microscopic diversity,and the biological evolution is driven by the amplification of biodiversity.Forming high hierarchies is the most important way for the amplification and brings out spontaneously three kinds of selection.This theory has some positive cultural meanings.

  2. Biological indices of lead exposure in relation to heavy metal residues in sediment and Biota from Prickly Pear Creek and Lake Helena, Montana

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — Biotic and sediment samples were collected from Lake Helena and Prickly Pear Creek upstream and downstream of the East Helena Smelter Superfund Site to assess metals...

  3. 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.

  4. 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.

  5. 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.

  6. Expression of viral EPS-depolymerase reduces fire blight susceptibility in transgenic pear.

    Science.gov (United States)

    Malnoy, Mickaël; Faize, Mohamed; Venisse, Jean-Stéphane; Geider, Klaus; Chevreau, Elisabeth

    2005-02-01

    Erwinia amylovora is the causal agent of fire blight of Maloideae. One of the main pathogenicity factors of this bacterium is the exopolysaccharide (EPS) of its capsule. In this paper, we used genetic transformation tools to constitutively express an EPS-depolymerase transgene in the pear (Pyrus communis L.) cv. Passe Crassane with the aim of decreasing its high susceptibility to fire blight. Expression of the depolymerase gene in 15 independent transgenic clones led, on average, to low depolymerase activity, although relatively high expression was observed at the transcriptional and translational levels. Only two of the transgenic clones (9X and 10M) consistently showed a decrease in fire blight susceptibility in vitro and in the greenhouse. These clones were also among the highest expressers of depolymerase at the RNA and enzyme activity levels. The correlation observed among all transgenic clones between depolymerase expression and fire blight resistance suggested the potential of this strategy.

  7. Biochemical and nutritional characterization of three prickly pear species with different ripening behavior.

    Science.gov (United States)

    Hernández-Pérez, Talia; Carrillo-López, Armando; Guevara-Lara, Fidel; Cruz-Hernández, Andrés; Paredes-López, Octavio

    2005-12-01

    Biochemical and nutritional changes were studied during the ripening process of three Opuntia morphospecies with different ripening behavior: Naranjona (O. ficus-indica), Blanca Cristalina (Opuntia sp.), and Esmeralda (Opuntia sp.) of early, early-intermediate, and intermediate-late ripening, respectively. In loss of fresh weight, Naranjona showed the highest values, while in Blanca Cristalina and Esmeralda, a discrete weight loss was found. No significant differences were found among morphospecies in soluble solids, total titratable acidity and pH during the postharvest days. Blanca Cristalina and Esmeralda showed an increase in the content of carotenoids, while these diminished in Naranjona. The cell wall enzymes evaluated showed particular behaviors during the ripening of each morphospecies suggesting a fine biochemical control and not a clear relationship between fruit softening and enzyme activity. This study provides basic information on prickly pear ripening, in order to understand this process for its control and for improving shelf life.

  8. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates.

    Science.gov (United States)

    Folmer, O; Black, M; Hoeh, W; Lutz, R; Vrijenhoek, R

    1994-10-01

    We describe "universal" DNA primers for polymerase chain reaction (PCR) amplification of a 710-bp fragment of the mitochondrial cytochrome c oxidase subunit I gene (COI) from 11 invertebrate phyla: Echinodermata, Mollusca, Annelida, Pogonophora, Arthropoda, Nemertinea, Echiura, Sipuncula, Platyhelminthes, Tardigrada, and Coelenterata, as well as the putative phylum Vestimentifera. Preliminary comparisons revealed that these COI primers generate informative sequences for phylogenetic analyses at the species and higher taxonomic levels.

  9. Quantitation of viral load using real-time amplification techniques

    NARCIS (Netherlands)

    Niesters, H G

    2001-01-01

    Real-time PCR amplification techniques are currently used to determine the viral load in clinical samples for an increasing number of targets. Real-time PCR reduces the time necessary to generate results after amplification. In-house developed PCR and nucleic acid sequence-based amplification (NASBA

  10. Isothermal DNA amplification in bioanalysis: strategies and applications.

    Science.gov (United States)

    Kim, Joonyul; Easley, Christopher J

    2011-01-01

    Isothermal DNA amplification is an alternative to PCR-based amplification for point-of-care diagnosis. Since the early 1990s, the approach has been refined into a simple, rapid and cost-effective tool by means of several distinct strategies. Input signals have been diversified from DNA to RNA, protein or small organic molecules by translating these signals into input DNA before amplification, thus allowing assays on various classes of biomolecules. In situ detection of single biomolecules has been achieved using an isothermal method, leveraging localized signal amplification in an intact specimen. A few pioneering studies to develop a homogenous isothermal protein assay have successfully translated structure-switching of a probe upon target binding into input DNA for isothermal amplification. In addition to the detection of specific targets, isothermal methods have made whole-genome amplification of single cells possible owing to the unbiased, linear nature of the amplification process as well as the large size of amplified products given by ϕ29 DNA polymerase. These applications have been devised with the four isothermal amplification strategies covered in this review: strand-displacement amplification, rolling circle amplification, helicase-dependent amplification and recombinase polymerase amplification.

  11. Plasmonic Terahertz Amplification in Graphene-Based Asymmetric Hyperbolic Metamaterial

    Directory of Open Access Journals (Sweden)

    Igor Nefedov

    2015-05-01

    Full Text Available We propose and theoretically explore terahertz amplification, based on stimulated generation of plasmons in graphene asymmetric hyperbolic metamaterials (AHMM, strongly coupled to terahertz radiation. In contrast to the terahertz amplification in resonant nanocavities, AHMM provides a wide-band THz amplification without any reflection in optically thin graphene multilayers.

  12. Plasmonic Terahertz Amplification in Graphene-Based Asymmetric Hyperbolic Metamaterial

    OpenAIRE

    Igor Nefedov; Leonid Melnikov

    2015-01-01

    We propose and theoretically explore terahertz amplification, based on stimulated generation of plasmons in graphene asymmetric hyperbolic metamaterials (AHMM), strongly coupled to terahertz radiation. In contrast to the terahertz amplification in resonant nanocavities, AHMM provides a wide-band THz amplification without any reflection in optically thin graphene multilayers.

  13. Adventitious shoot regeneration from the leaves of some pear varieties (Pyrus spp.) grown in vitro

    Institute of Scientific and Technical Information of China (English)

    Bharat Kumar POUDYAL; Yuxing ZHANG; Guoqiang DU

    2008-01-01

    The pear (Pyrus spp.) is one of the most important temperate fruit crops. A complete protocol for adventitious shoot regeneration was developed from the leaves of four pear varieties grown in vitro: Abbe Fetel, Yali, Packham's Triumph and Aikansui, and the Chinese rootstock variety Dull. Shoot explants were collected from the field and cultured in vitro in Murashige and acid (IBA). After four weeks, leaf explants of all 5 varieties grown in vitro were excised and cultured in MS cultures were maintained in darkness for 21 days for shoot induction in the shoot induction medium (IM), then transferred to the shoot expression medium (EM) in room at (25±2)℃ under a 16/8 h light/dark photoperiod regime for 8 weeks. Finally, the shoots were transferred to the MS shoot elongation medium (SEM) supplemented gibberellic acid (GA3). A combination of TDZ and NAA had a significant effect on the number of shoot regenera-tions in all 5 tested varieties. The maximum mean number of shoots and maximum number of shoots per leaf obtained from Yali variety were 11.8 (P≤0.001) and 22, followed by Aikansui with 6.6 (P≤0.001) and 4.6, and Duff with 8 (P≤0.001) and 12, all arising from the For Packham's Triumph and Abbe Fetel, the maximum mean number of shoots and maximum number of shoots per leaf were 5.6 (P≤0.001), 4.8 and 8 (P≤0.001), and 11, which produced significantly higher adventitious shoots problems associated with shoot proliferation and regenera-tion were also observed and discussed in this paper.

  14. Stem Water Potential Monitoring in Pear Orchards through WorldView-2 Multispectral Imagery

    Directory of Open Access Journals (Sweden)

    Jonathan Van Beek

    2013-12-01

    Full Text Available Remote sensing can provide good alternatives for traditional in situ water status measurements in orchard crops, such as stem water potential (Ψstem. However, the heterogeneity of these cropping systems causes significant differences with regards to remote sensing products within one orchard and between orchards. In this study, robust spectral indicators of Ψstem were sought after, independent of sensor viewing geometry, orchard architecture and management. To this end, Ψstem was monitored throughout three consecutive growing seasons in (deficit irrigated and rainfed pear orchards and related to spectral observations of leaves, canopies and WorldView-2 imagery. On a leaf and canopy level, high correlations were observed between the shortwave infrared reflectance and in situ measured Ψstem. Additionally, for canopy measurements, visible and near-infrared wavelengths (R530/R600, R530/R700 and R720/R800 showed significant correlations. Therefore, the Red-edge Normalized Difference Vegetation Index (ReNDVI was applied on fully sunlit satellite imagery and found strongly related with Ψstem (R2 = 0.47; RMSE = 0.36 MPa, undoubtedly showing the potential of WorldView-2 to monitor water stress in pear orchards. The relationship between ReNDVI and Ψstem was independent of management, irrigation setup, phenology and environmental conditions. In addition, results showed that this relation was also independent of off-nadir viewing angle and almost independent of viewing geometry, as the correlation decreased after the inclusion of fully shaded scenes. With further research focusing on issues related to viewing geometry and shadows, high spatial water status monitoring with space borne remote sensing is achievable.

  15. THE ROLE OF MINERAL NUTRITION ON YIELDS AND FRUIT QUALITY IN GRAPEVINE, PEAR AND APPLE

    Directory of Open Access Journals (Sweden)

    GUSTAVO BRUNETTO

    2015-12-01

    Full Text Available ABSTRACT Fertilization of temperate fruit trees, such as grapevine ( Vitis spp., apple ( Malus domestica, and pear ( Pyrus communis is an important tool to achive maximum yield and fruit quality. Fertilizers are provided when soil fertility does not allow trees to express their genetic potential, and time and rate of application should be scheduled to promote fruit quality. Grapevine berries, must and wine quality are affected principally by N, that regulate the synthesis of some important compounds, such as anthocyanins, which are responsible for coloring of the must and the wine. Fermenation of the must may stop in grapes with low concentration of N because N is requested in high amount by yeasts. An N excess may increase the pulp to peel ratio, diluting the concentration of anthocyanins and promoting the migration of anthocyanins from berries to the growing plant organs; a decrease of grape juice soluble solid concentration is also expected because of an increase in vegetative growth. Potassium is also important for wine quality contributing to adequate berry maturation, concentration of sugars, synthesis of phenols and the regulation of pH and acidity. In apple and pear, Ca and K are important for fruit quality and storage. Potassium is the most important component of fruit, however, any excess should be avoided and an adequate K:Ca balance should be achieved. Adequate concentration of Ca in the fruit prevents pre- and post-harvest fruit disorders and, at the same time, increases tolerance to pathogens. Although N promotes adequate growth soil N availability should be monitored to avoid excessive N uptake that may decrease fruit skin color and storability.

  16. Factors involved in alleviating water stress by partial crop removal in pear trees.

    Science.gov (United States)

    Marsal, Jordi; Mata, Merce; Arbones, Amadeu; Del Campo, Jesus; Girona, Joan; Lopez, Gerardo

    2008-09-01

    We studied the relief of water stress associated with fruit thinning in pear (Pyrus communis L.) trees during drought to determine what mechanisms, other than stomatal adjustment, were involved. Combinations of control irrigation (equal to crop water use less effective rainfall) and deficit irrigation (equal to 20% of control irrigation), fruit load (unthinned and thinned to 40 fruits per tree) and root pruning (pruned and unpruned) treatments were applied to pear (cv. 'Conference') trees during Stage II of fruit development. Daily patterns of midday stem water potential (Psi(stem)) and leaf conductance to water vapor (g(l)) of deficit-irrigated trees differed after fruit thinning. In response to fruit thinning, gl progressively declined with water stress until 30 days after fruit thinning and then leveled off, whereas the effects of decreased fruit load on Psi(stem) peaked 30-40 days after fruit thinning and then tended to decline. Soil water depletion was significantly correlated with fruit load during drought. Our results indicate that stomatal adjustment and the resulting soil water conservation were the factors determining the Psi(stem) response to fruit thinning. However, these factors could not explain differences in daily patterns between g(l) and Psi(stem) after fruit thinning. In all cases, effects of root pruning treatments on Psi(stem) in deficit-irrigated trees were transitory (Psi(stem) recovered from root pruning in less than 30 days), but the recovery of Psi(stem) after root pruning was faster in trees with low fruit loads. This behavior is compatible with the concept that the water balance (reflected by Psi(stem) values) was better in trees with low fruit loads compared with unthinned trees, perhaps because more carbon was available for root growth. Thus, a root growth component is hypothesized as a mechanism to explain the bimodal Psi(stem) response to fruit thinning during drought.

  17. Erwinia piriflorinigrans sp. nov., a novel pathogen that causes necrosis of pear blossoms.

    Science.gov (United States)

    López, María M; Roselló, Montserrat; Llop, Pablo; Ferrer, Sergi; Christen, Richard; Gardan, Louis

    2011-03-01

    Eight Erwinia strains, isolated from necrotic pear blossoms in València, Spain, were compared with reference strains of Erwinia amylovora and Erwinia pyrifoliae, both of which are pathogenic to species of pear tree, and to other species of the family Enterobacteriaceae using a polyphasic approach. Phenotypic analyses clustered the novel isolates into one phenon, distinct from other species of the genus Erwinia, showing that the novel isolates constituted a homogeneous phenotypic group. Rep-PCR profiles, PCR products obtained with different pairs of primers and plasmid contents determined by restriction analysis showed differences between the novel strains and reference strains of E. amylovora and E. pyrifoliae. Phylogenetic analysis of 16S rRNA, gpd and recA gene sequences showed that the eight novel strains could not be assigned to any recognized species. On the basis of DNA-DNA hybridization studies, the novel isolates constituted a single group with relatedness values of 87-100  % to the designated type strain of the group, CFBP 5888(T). Depending on the method used, strain CFBP 5888(T) showed DNA-DNA relatedness values of between 22.7 and 50  % to strains of the closely related species E. amylovora and E. tasmaniensis. The DNA G+C contents of two of the novel strains, CFBP 5888(T) and CFBP 5883, were 51.1 and 50.5 mol%, respectively. On the basis of these and previous results, the novel isolates represent a novel species of the genus Erwinia, for which the name Erwinia piriflorinigrans sp. nov. is proposed. The type strain is CFBP 5888(T) (=CECT 7348(T)).

  18. Evaluation of Susceptibility of Different Pear Hybrid Populations to Fire Blight (Erwinia amylovora

    Directory of Open Access Journals (Sweden)

    Yasemin EVRENOSOĞLU

    2011-05-01

    Full Text Available Fire blight disease caused by pathogenic bacterium Erwinia amylovora, is the serious disease of pear, and there is not a certain chemical management against this disease except antibiotic-type compounds such as streptomycin. It is very important to improve new fire blight resistant cultivars in case of integrated disease management. With this purpose, different crosses have been made between Pyrus communis varieties that have good fruit characteristics and resistant cultigens. Besides, self and open pollination treatments have been carried out in maternal plants. The disease resistance level of the hybrids obtained from these combinations was determined by artificial inoculations by Erwinia amylovora in greenhouse conditions. A total of 3284 hybrids were inoculated, and 2631 of them survived and were distributed to different susceptibility classes. 19.88% of the inoculated hybrids was killed by Erwinia amylovora. Total distribution of the hybrids to susceptibility classes was as 6.18% in class “A- slightly susceptible”, 3.11% in class “B- less susceptible”, 8.89% in class “C- mid-susceptible”, 20.28% in class “D- susceptible”, and 61.54% in class “E- very susceptible”. Majority of class “A- slightly susceptible” hybrids were obtained from ‘Magness’ x ‘Ankara’ combination. ‘Kieffer’ x ‘Santa Maria’, ‘Kieffer’ open pollination, ‘Magness’ x ‘Akça’, ‘Magness’ x ‘Kieffer’, ‘Magness’ x ‘Santa Maria’, ‘Mustafa Bey’ x ‘Moonglow’ treatments displayed good results with respect to “A- slightly susceptible” character. It is very important to evaluate these hybrid pear populations through different fruit and tree characteristics in the future.

  19. 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.

  20. Rehabilitation of Degraded Rangeland in Drylands by Prickly Pear (Opuntia ficus-indica L.) Plantations: Effect on Soil and Spontaneous Vegetation

    OpenAIRE

    Souad Neffar; Haroun Chenchouni; Arifa Beddiar; Noureddine Redjel

    2013-01-01

    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 ...

  1. Development of Fermented Beverage of Pear Juice%发酵型梨汁饮料的研制

    Institute of Scientific and Technical Information of China (English)

    王宁; 艾学东

    2015-01-01

    研究以梨汁和酒精为主要原料,经醋酸发酵后的原液中加入果葡糖浆、蜂蜜等调制发酵型梨汁饮料。利用单因素分析,探讨了梨汁糖度、酒精含量对发酵梨汁原液的影响。结果表明,糖度为6°Brix、酒精添加量1.5g/100mL时发酵梨汁原液效果最好;采用正交试验,以总酸为有效成分指标,研究了有氧发酵的最佳工艺条件。结果表明,菌种接种量10%、发酵温度32℃、发酵时间72h为最佳有氧发酵工艺;确定发酵型梨汁饮料的最佳配方为:发酵梨汁原液80%、果葡糖浆7.5%、蜂蜜1%。%Pear juice and alcohol are used as raw material to produce pear juice beverage by adding fructose syrup and honey into acetic fermentation solution. By single factor analysis,effect of sugar level and alcohol content on fermentation of pear juice concentrate was discussed. The results showed that when sugar is in 6°Brix and alcohol fermentation of 1.5g/100mL,fermentation of pear juice solution is optimal. Using orthogonal experiment and index of total acid as active ingredients,optimum process conditions of aerobic fermentation was studied. The results showed that the species quantity of 10%,32℃fermentation temperature,fermentation time 72h is the best aerobic fermentation process;By means of orthogonal experiment,best formula for the fermentation type of pear juice drink was determined: fermenting pear juice concentrate 80%,fructose syrup 7.5%,and honey 1%.

  2. 洋葱雪梨复合饮料的研制%Compound Beverage of Onion and Pear

    Institute of Scientific and Technical Information of China (English)

    李素; 党超; 刘晨; 祝传望; 王跃猛

    2014-01-01

    为了提高洋葱和雪梨的利用率并增加其附加值,进一步拓宽洋葱和雪梨的市场,以洋葱和雪梨为主要原材料,通过正交试验和感官评价的方法确定最佳生产工艺及配方,复合形成一种风味独特、营养丰富的洋葱雪梨复合饮料。先将洋葱和雪梨分别按料液比为1∶2的比例加水榨汁,并添加0.2%的异抗坏血酸护色,然后按雪梨汁∶洋葱汁5∶1、白砂糖3%、黄原胶0.5%、柠檬酸0.5%的配方制成洋葱雪梨饮料,产品呈乳白色,组织状态均匀,洋葱和雪梨复合出独特的风味,清香爽口。%Compound beverage of onion and pear was made to improve utilization efficiency and increase the additional value of onions and pear, also, broaden the market of onion and pear. With onion and pear as main raw materials, the processing technology and formula were studied by the method of orthogonal test and sensory evaluation. The optimum formula of the compound beverage was that onion juice: pear juice = 5∶1, sugar 3%, xanthan gum 0.5%, citric acid 0.5%, the onion juice and pear juice were made with the water than expected for the 1∶2, added 0.2% ascorbic acid to protect the color. The beverage was of rich nutrition, white, and with unique compound flavor.

  3. Product length, dye choice, and detection chemistry in the bead-emulsion amplification of millions of single DNA molecules in parallel.

    Science.gov (United States)

    Tiemann-Boege, Irene; Curtis, Christina; Shinde, Deepali N; Goodman, Daniel B; Tavaré, Simon; Arnheim, Norman

    2009-07-15

    The amplification of millions of single molecules in parallel can be performed on microscopic magnetic beads that are contained in aqueous compartments of an oil-buffer emulsion. These bead-emulsion amplification (BEA) reactions result in beads that are covered by almost-identical copies derived from a single template. The post-amplification analysis is performed using different fluorophore-labeled probes. We have identified BEA reaction conditions that efficiently produce longer amplicons of up to 450 base pairs. These conditions include the use of a Titanium Taq amplification system. Second, we explored alternate fluorophores coupled to probes for post-PCR DNA analysis. We demonstrate that four different Alexa fluorophores can be used simultaneously with extremely low crosstalk. Finally, we developed an allele-specific extension chemistry that is based on Alexa dyes to query individual nucleotides of the amplified material that is both highly efficient and specific.

  4. Rapid detection of Salmonella Typhi by loop-mediated isothermal amplification (LAMP) method.

    Science.gov (United States)

    Abdullah, J; Saffie, N; Sjasri, F A R; Husin, A; Abdul-Rahman, Z; Ismail, A; Aziah, I; Mohamed, M

    2014-01-01

    An in-house loop-mediated isothermal amplification (LAMP) reaction was established and evaluated for sensitivity and specificity in detecting the presence of Salmonella Typhi (S. Typhi) isolates from Kelantan, Malaysia. Three sets of primers consisting of two outer and 4 inner were designed based on locus STBHUCCB_38510 of chaperone PapD of S. Typhi genes. The reaction was optimised using genomic DNA of S. Typhi ATCC7251 as the template. The products were visualised directly by colour changes of the reaction. Positive results were indicated by green fluorescence and negative by orange colour. The test was further evaluated for specificity, sensitivity and application on field samples. The results were compared with those obtained by gold standard culture method and Polymerase Chain Reaction (PCR). This method was highly specific and -10 times more sensitive in detecting S. Typhi compared to the optimised conventional polymerase chain reaction (PCR) method.

  5. On-Chip Isothermal Nucleic Acid Amplification on Flow-Based Chemiluminescence Microarray Analysis Platform for the Detection of Viruses and Bacteria.

    Science.gov (United States)

    Kunze, A; Dilcher, M; Abd El Wahed, A; Hufert, F; Niessner, R; Seidel, M

    2016-01-01

    This work presents an on-chip isothermal nucleic acid amplification test (iNAAT) for the multiplex amplification and detection of viral and bacterial DNA by a flow-based chemiluminescence microarray. In a principle study, on-chip recombinase polymerase amplification (RPA) on defined spots of a DNA microarray was used to spatially separate the amplification reaction of DNA from two viruses (Human adenovirus 41, Phi X 174) and the bacterium Enterococcus faecalis, which are relevant for water hygiene. By establishing the developed assay on the microarray analysis platform MCR 3, the automation of isothermal multiplex-amplification (39 °C, 40 min) and subsequent detection by chemiluminescence imaging was realized. Within 48 min, the microbes could be identified by the spot position on the microarray while the generated chemiluminescence signal correlated with the amount of applied microbe DNA. The limit of detection (LOD) determined for HAdV 41, Phi X 174, and E. faecalis was 35 GU/μL, 1 GU/μL, and 5 × 10(3) GU/μL (genomic units), which is comparable to the sensitivity reported for qPCR analysis, respectively. Moreover the simultaneous amplification and detection of DNA from all three microbes was possible. The presented assay shows that complex enzymatic reactions like an isothermal amplification can be performed in an easy-to-use experimental setup. Furthermore, iNAATs can be potent candidates for multipathogen detection in clinical, food, or environmental samples in routine or field monitoring approaches.

  6. Gravito-magnetic amplification in cosmology

    CERN Document Server

    Tsagas, Christos G

    2009-01-01

    Magnetic fields interact with gravitational waves in various ways. We consider the coupling between the Weyl and the Maxwell fields in cosmology and study the effects of the former on the latter. The approach is fully analytical and the results are gauge-invariant. We show that the nature and the outcome of the gravito-magnetic interaction depends on the electric properties of the cosmic medium. When the conductivity is high, gravitational waves reduce the standard (adiabatic) decay rate of the B-field, leading to its superadiabatic amplification. In poorly conductive environments, on the other hand, Weyl-curvature distortions can result into the resonant amplification of large-scale cosmological magnetic fields. Driven by the gravitational waves, these B-fields oscillate with an amplitude that is found to diverge when the wavelengths of the two sources coincide. We present technical and physical aspects of the gravito-magnetic interaction and discuss its potential implications.

  7. 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.

  8. Amplification of postwildfire peak flow by debris

    Science.gov (United States)

    Kean, J. W.; McGuire, L. A.; Rengers, F. K.; Smith, J. B.; Staley, D. M.

    2016-08-01

    In burned steeplands, the peak depth and discharge of postwildfire runoff can substantially increase from the addition of debris. Yet methods to estimate the increase over water flow are lacking. We quantified the potential amplification of peak stage and discharge using video observations of postwildfire runoff, compiled data on postwildfire peak flow (Qp), and a physically based model. Comparison of flood and debris flow data with similar distributions in drainage area (A) and rainfall intensity (I) showed that the median runoff coefficient (C = Qp/AI) of debris flows is 50 times greater than that of floods. The striking increase in Qp can be explained using a fully predictive model that describes the additional flow resistance caused by the emergence of coarse-grained surge fronts. The model provides estimates of the amplification of peak depth, discharge, and shear stress needed for assessing postwildfire hazards and constraining models of bedrock incision.

  9. An integrated portable hand-held analyser for real-time isothermal nucleic acid amplification.

    Science.gov (United States)

    Smith, Matthew C; Steimle, George; Ivanov, Stan; Holly, Mark; Fries, David P

    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 (R2 = 0.948) to fluorescein gradients ranging from 0.5 to 10 microM 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.

  10. Detection of total and hemolysin-producing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tl, tdh and trh.

    Science.gov (United States)

    Bej, A K; Patterson, D P; Brasher, C W; Vickery, M C; Jones, D D; Kaysner, C A

    1999-06-01

    Vibrio parahaemolyticus is an important human pathogen which can cause gastroenteritis when consumed in raw or partially-cooked seafood. A multiplex PCR amplification-based detection of total and virulent strains of V. parahaemolyticus was developed by targeting thermolabile hemolysin encoded by tl, thermostable direct hemolysin encoded by tdh, and thermostable direct hemolysin-related trh genes. Following optimization using oligonucleotide primers targeting tl, tdh and trh genes, the multiplex PCR was applied to V. parahaemolyticus from 27 clinical, 43 seafood, 15 environmental, 7 strains obtained from various laboratories and 19 from oyster plants. All 111 V. parahaemolyticus isolates showed PCR amplification of the tl gene; however, only 60 isolates showed amplification of tdh, and 43 isolates showed amplification of the trh gene. Also, 18 strains showed amplification of the tdh gene, but these strains did not show amplification of the trh gene. However, one strain exhibited amplification for the trh but not the tdh gene, suggesting both genes need to be targeted in a PCR amplification reaction to detect all hemolysin-producing strains of this pathogen. The multiplex PCR approach was successfully used to detect various strains of V parahaemolyticus in seeded oyster tissue homogenate. Sensitivity of detection for all three target gene segments was at least between 10(1)-10(2) cfu per 10 g of alkaline peptone water enriched seeded oyster tissue homogenate. This high level of sensitivity of detection of this pathogen within 8 h of pre-enrichment is well within the action level (10(4) cfu per 1 g of shell stock) suggested by the National Seafood Sanitation Program guideline. Compared to conventional microbiological culture methods, this multiplex PCR approach is rapid and reliable for accomplishing a comprehensive detection of V. parahaemolyticus in shellfish.

  11. Internal entanglement amplification by external interactions

    OpenAIRE

    2007-01-01

    We propose a scheme to control the level of entanglement between two fixed spin-1/2 systems by interaction with a third particle. For specific designs, entanglement is shown to be "pumped" into the system from the surroundings even when the spin-spin interaction within the system is small or nonexistent. The effect of the external particle on the system is introduced by including a dynamic spinor in the Hamiltonian. Controlled amplification of the internal entanglement to its maximum value is...

  12. Erwinia amylovora loop-mediated isothermal amplification (LAMP) assay for rapid pathogen detection and on-site diagnosis of fire blight.

    Science.gov (United States)

    Bühlmann, Andreas; Pothier, Joël F; Rezzonico, Fabio; Smits, Theo H M; Andreou, Michael; Boonham, Neil; Duffy, Brion; Frey, Jürg E

    2013-03-01

    Several molecular methods have been developed for the detection of Erwinia amylovora, the causal agent of fire blight in pear and apple, but none are truly applicable for on-site use in the field. We developed a fast, reliable and field applicable detection method using a novel target on the E. amylovora chromosome that we identified by applying a comparative genomic pipeline. The target coding sequences (CDSs) are both uniquely specific for and all-inclusive of E. amylovora genotypes. This avoids potential false negatives that can occur with most commonly used methods based on amplification of plasmid gene targets, which can vary among strains. Loop-mediated isothermal AMPlification (LAMP) with OptiGene Genie II chemistry and instrumentation proved to be an exceptionally rapid (under 15 min) and robust method for detecting E. amylovora in orchards, as well as simple to use in the plant diagnostic laboratory. Comparative validation results using plant samples from inoculated greenhouse trials and from natural field infections (of regional and temporal diverse origin) showed that our LAMP had an equivalent or greater performance regarding sensitivity, specificity, speed and simplicity than real-time PCR (TaqMan), other LAMP assays, immunoassays and plating, demonstrating its utility for routine testing.

  13. 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.

  14. The Role of DNA Amplification and Cultural Growth in Complicated Acute Appendicitis

    Science.gov (United States)

    Tocchioni, Francesca; Tani, Chiara; Bartolini, Laura; Moriondo, Maria; Nieddu, Francesco; Pecile, Patrizia; Azzari, Chiara; Messineo, Antonio; Ghionzoli, Marco

    2016-01-01

    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. PMID:27777701

  15. Integrated platform with magnetic purification and rolling circular amplification for sensitive fluorescent detection of ochratoxin A.

    Science.gov (United States)

    Yao, Li; Chen, Yinji; Teng, Jun; Zheng, Wanli; Wu, Jingjing; Adeloju, Samuel B; Pan, Daodong; Chen, Wei

    2015-12-15

    In this article, we report the detection of ochratoxin A (OTA) with excellent sensitivity with the two-aspect signal amplification treatments. Combining the unique property of magnetic nanoparticles and the high efficiency of the in vitro amplification of rolling circular amplification (RCA), the competitive sensing protocol for ultrasensitive detection of OTA was achieved in about 80 min. The excellent magnetic separation treatment could effectively avoid the interference of background fluorescent noise in the sensing system while the RCA could tremendously increase the hybridization sequence for the quantum dot labeled probes and further increase the sensing response signal. Afterwards, two factors affecting the final detection limit, concentration of RCA components and RCA reaction time, were all systematically optimized for the best sensing performance. The response of the optimized protocol for OTA detection is highly linear over the wider range from 10(-3) to 10 ppb, which is 3 orders improvement in sensing range, and the limit of detection is calculated to be as low as 0.13 ppt, which is 10,000 folds improvement compared with the traditional methods. More importantly, given the selected aptamer, this universal signal amplification protocol could be widely applied to other fields by just change the recognition sequence of the aptamer.

  16. Fermentation and Aroma Analysis of Jingbai Pear Wine%京白梨酒发酵与香气分析

    Institute of Scientific and Technical Information of China (English)

    宋柬; 李德美; 邓小明; 张莉华; 陈尚武; 马会勤

    2012-01-01

    在京白梨酒发酵工艺研究的基础上,利用固相微萃取-气相色谱-嗅辨-质谱法进一步对京白梨酒特征性香气成分进行了分析.接种发酵前,分别采用浓缩梨汁、白砂糖、秋梨膏调整京白梨汁的含糖量,对比原汁和果渣的发酵方式、感官评价结果、挥发性成分种类和经济成本等综合因素,最终确定添加白砂糖来调整京白梨汁的糖度到16°Brix.结合前期的实验条件确定了京白梨酒的中试工艺参数.京白梨酒可能的特征香气物质包括:乙酸异戊酯、己酸乙酯、乙酸己酯、辛酸甲酯、丁二酸二乙酯、辛酸乙酯、乙酸苯乙酯、9-癸烯酸乙酯、癸酸乙酯、异戊醇、正己醇、β-苯乙醇和紫罗兰酮.本研究通过不同发酵处理对香气成分的分析,为进一步提高京白梨酒的品质提供了新的依据.%The aromatic compounds in perry were evaluated by combining SPME with GC - MS and GC - O methods on the basis of fermentation in Jingbai pear. Before fermentation, the sugar degree of Jingbai pear juice was adjusted by pear juice concentrate, granulated sugar and autumn pear grease separately- It was confirmed that pear wine adjusted to 16癇rix by granulated sugar, gained the best sensory quality and more affluent volatile components, comparing to the raw juice and pomace. The Jingbai pear wine pilot scale fermentation was carried on based on the laboratory parameters. The key aroma components included l-butanol, 3-methyl-, acetate; hexanoic acid, ethyl ester; acetic acid, hexyl ester; octanoic acid, methyl ester; butanedioic acid, diethyl ester; octanoic acid, ethyl ester; acetic acid, 2-phenylethyl ester; ethyl 9-decenoate; decanoic acid, ethyl ester; l-butanol, 3-methyl-; 1-hexanol; phenylethyl alcohol; 3-buten-2-one, 4- (2,6,6- trimethyl-2- cyclohexen-1-yl). These aroma components were compared in different pear wines. The results illustrated a new way for the quality Jingbai pear wine improvement

  17. Pilot-scale production of cloudy juice from low-quality pear fruit under low-oxygen conditions.

    Science.gov (United States)

    De Paepe, Domien; Coudijzer, Katleen; Noten, Bart; Valkenborg, Dirk; Servaes, Kelly; De Loose, Marc; Diels, Ludo; Voorspoels, Stefan; Van Droogenbroeck, Bart

    2015-04-15

    In this study, a process for the production of premium quality yellowish, cloudy pear juice from low-quality fruit under low-oxygen conditions was developed. The production process consisted of (1) shredding, (2) pressing with spiral-filter technology including a vacuumised extraction cell, (3) holding in an inert gas buffer tank, (4) pasteurisation, (5) and refrigerated storage. First, the system parameters of a spiral-filter press were optimised with the aim of producing a yellowish, cloudy pear juice with the highest possible juice yield. A maximum juice yield of 78% could be obtained. Enzymatic browning during juice extraction could be suppressed as a result of the fast processing and the low air (oxygen) levels in the extraction chamber of the spiral-filter press. Furthermore, we observed that instantaneous pasteurisation at 107 °C for 6s, subsequent aluminium laminate packaging and cold storage had only a minimum effect on the phenolic composition.

  18. Study on Water Consumption Rule of Potted Pear%盆栽梨树的耗水规律

    Institute of Scientific and Technical Information of China (English)

    程福厚; 赵岩丽; 张纪英; 宋惠月; 赵志军; 王庆江

    2011-01-01

    In order to study the water consumption characteristics of pear in different soil moisture conditions and different seasons, using potted cultural Huangguan pear of four-year old as the experiment materials, water consumption of different soil water status pear (Pyrus bretschneidery Rehd) was studied by weighting method.The correlation coefficient between the consumption amount and the climatic element and area of leaf was analyzed. The results showed that adequate soil water treatment was significantly higher than moderate and severe drought treatment in water consumption. The water consumption during the canopy formation was small (0.25-0.58 L/day). Early June was the biggest (10.1 L/ten days) among every ten days. The summit in a day was at 10:00-14:00. With the decline in soil moisture, photosynthetic rate of pear was significantly lower in the mid growing season of pear, other factors were not significantly different. The regression analysis results showed that average air temperature was the biggest influence factor. There were distinguished positive correlation between the water consumption and average air temperature (P=0.0075), wind speed (P=0.0135), leaf area on pear (r=0.9516), distinguished negative correlation between the water consumption and air humidity (P=0.0462). It is suggested that development phase of pear, soil water, leaf area, air temperature, air humidity and wind speed are the bases to make the consumption water model of pear.%为探讨梨树在不同土壤水分条件下和不同季节的耗水规律,以4年生盆栽黄冠梨为试材,用称重法研究了黄冠梨在5-10月不同土壤水分条件下的日耗水量,分析了耗水量与气象因子及叶面积的相关性.结果表明:在适宜水分条件下梨树的耗水量显著大于中度亏缺和重度亏缺条件下的耗水量.梨树叶幕形成阶段日耗水量较小,在0.25~0.58 L/日.从旬耗水量的变化来看,以6月上旬耗水量最大,达到10.1L/旬,一

  19. Analysis of six organophosphorus pesticide residues in apples and pears using cloud-point extraction coupled with HPLC-UV.

    Science.gov (United States)

    Zhang, Lijin; Chen, Fang; Zhang, Wenhuan; Pan, Canping

    2014-01-01

    A cloud-point extraction (CPE) method with Triton X-114 has been developed for analysis of six organophosphorus pesticides (OPPs) in apples and pears. In this CPE procedure, the effects of the surfactant volume, mass of sodium chloride, equilibrium temperature, equilibrium time, and pH on the extraction procedure were investigated. Under the optimal CPE conditions, the analytes were enriched 20-fold and the LODs dropped to 0.44-5.20 microg/kg. Furthermore, the proposed extraction method was validated by the correlation coefficient (R2) of the calibration curve, repeatability (RSD, n = 6), and fortified recoveries, which were 0.9967-0.9993, 2.7-6.5, and 74.7-104.5%, respectively. Based on these results, it could be concluded that the proposed CPE method with Triton X-114 was suitable for the effective extraction and enrichment of OPP residues in the apple and pear samples.

  20. γ-Aminobutyric acid induces resistance against Penicillium expansum by priming of defence responses in pear fruit.

    Science.gov (United States)

    Yu, Chen; Zeng, Lizhen; Sheng, Kuang; Chen, Fangxia; Zhou, Tao; Zheng, Xiaodong; Yu, Ting

    2014-09-15

    The results from this study showed that treatment with γ-aminobutyric acid (GABA), at 100-1000 μg/ml, induced strong resistance against blue mould rot caused by Penicillium expansum in pear fruit. Moreover, the activities of five defence-related enzymes (including chitinase, β-1,3-glucanase, phenylalnine ammonialyase, peroxidase and polyphenol oxidase) and the expression of these corresponding genes were markedly and/or promptly enhanced in the treatment with GABA and inoculation with P. expansum compared with those that were treated with GABA or inoculated with pathogen alone. In addition, the treatment of pear with GABA had little adverse effect on the edible quality of the fruit. To the best of our knowledge, this is the first report that GABA can effectively reduce fungal disease of harvested fruit. Its mechanisms may be closely correlated with the induction of fruit resistance by priming activation and expression of defence-related enzymes and genes upon challenge with pathogen.

  1. 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

  2. 块菌梨醋加工工艺研究%Study on Processing Technology of Truffle Pear Vinegar

    Institute of Scientific and Technical Information of China (English)

    王艳丽; 邓杰; 卫春会; 钟姝霞; 祝云飞; 黄治国

    2016-01-01

    以梨为原料,采用发酵方法制取梨醋,分析发酵过程中糖度、酒精度、酸度的变化规律,结果表明:酵母发酵过程中随时间的增加,梨汁的糖度下降,酒精度上升;醋酸发酵过程中随时间增加,酒精度逐渐减少,酸度不断增加。调配采用正交试验,结果表明:最佳的调配配方为梨醋20 mL、块菌提取液20 mL、蔗糖4 g、柠檬酸0.06 g,调配出的复合饮料色泽亮丽,具有块菌特殊香气。%In this paper,pear is used as raw material to make pear vinegar by fermentation method, analyze the variation of sugar degree,alcoholic strength and acidity in the process of fermentation,the results show that with the increase of time,the sugar degree of pear juice declines and alcoholic strength rises.During the process of acetic acid fermentation,with the increase of time,the alcoholic strength declines gradually,the acidity rises continuously.The orthogonal experiment results show that under the conditions of pear vinegar of 20 mL,truffle extraction liquid of 20 mL,sucrose of 4 g, citric acid of 0.06 g,the compound beverage has bright color and special truffle aroma.

  3. Research on the Brand Development of Dangshan Pear%砀山酥梨品牌发展研究

    Institute of Scientific and Technical Information of China (English)

    胡月英

    2014-01-01

    本文简要阐述了砀山酥梨品牌的发展现状,较为深入地分析了砀山酥梨品牌发展中存在问题,提出了要提高砀山酥梨生产经营者的品牌意识、树立砀山酥梨品牌的绿色水果形象、加强砀山酥梨品牌整合营销传播、完善砀山酥梨的标准化体系提高产品标准化程度、健全品牌管理制度等品牌发展策略,以促进砀山酥梨品牌很好发展,为果农带来更多收益,带动地方经济快速发展。%This paper briefly expounded the recent situation of the brand development of Dangshan Pears,thor-oughly analyzed the problems in the development of the brand and raised that to improve the Dangshan Pears op-eratorˊs brand awareness,strengthen the integrated marketing communication,perfect the standardization system of Dangshan Pears and improve the degree of product standardization,we should complete the brand development strategy as the brand management system,in order to promote the development of Dangshan Pear brand,bring more income for orchardman,and drive the local economy to develop rapidly.

  4. Relationship between shelf-life and optical properties of Yuanhuang pear in the region of 400-1150 nm

    Science.gov (United States)

    He, Xueming; Fu, Xiaping; Rao, Xiuqin; Fang, Zhenhuan

    2016-05-01

    The main goals of this study are to investigate the potential of absorption coefficient for the prediction of water contents in `Yuanhuang' pear and analyze the relationship between the shelf-life and bulk optical properties in the range of 900-1050 nm. An automated integrating sphere (AIS) system was used to measure the total reflectance, total transmittance of pear flesh tissues in visible-Near infrared (Vis-NIR) range. These two measurements were used to estimate the absorption coefficient μa and reduced scattering coefficient μ's of pear samples by using an inverse adding doubling (IAD) light propagation model. The detection accuracy of the AIS system was verified by using both liquid (Intralipid-20% as scatterer) and solid phantom (TiO2 as scatterer, carbon black as absorber). The relative error of measurement of μ's of liquid phantom with four different concentration (0.5%,1%,1.5%,2%) at 632.8 nm, 751 nm, 833 nm are less than 10% except for 2% concentration at 833 nm, and the relative error of measurement μa and μ's of solid phantom at 525.4 nm, 632.1 nm, 710.3 nm and 780.1 nm are less than 5% except for the μa at 525.4 nm. A total of 140 samples were used to conduct the moisture measurement, and drying method was used. Predictive models for moisture content from μa data were constructed using partial least squares regression (PLSR). The coefficient of correlation of calibration set (Rc) and validation set (Rp) were 0.50 and 0.45 respectively. The relationship between the shelf-life and optical properties was analyzed by dividing pear samples into three categories according to the actual shelf-life, and calculating classification accuracy by using actual and calculated shelf-life grade.

  5. Pistil-function breakdown in a new S-allele of European pear, S21*, confers self-compatibility.

    Science.gov (United States)

    Sanzol, Javier

    2009-03-01

    European pear exhibits RNase-based gametophytic self-incompatibility controlled by the polymorphic S-locus. S-allele diversity of cultivars has been extensively investigated; however, no mutant alleles conferring self-compatibility have been reported. In this study, two European pear cultivars, 'Abugo' and 'Ceremeño', were classified as self-compatible after fruit/seed setting and pollen tube growth examination. S-genotyping through S-PCR and sequencing identified a new S-RNase allele in the two cultivars, with identical deduced amino acid sequence as S(21), but differing at the nucleotide level. Test-pollinations and analysis of descendants suggested that the new allele is a self-compatible pistil-mutated variant of S(21), so it was named S(21)*. S-genotypes assigned to 'Abugo' and 'Ceremeño' were S(10)S(21)* and S(21)*S(25) respectively, of which S(25) is a new functional S-allele of European pear. Reciprocal crosses between cultivars bearing S(21) and S(21)* indicated that both alleles exhibit the same pollen function; however, cultivars bearing S(21)* had impaired pistil-S function as they failed to reject either S(21) or S (21)* pollen. RT-PCR analysis showed absence of S(21)* -RNase gene expression in styles of 'Abugo' and 'Ceremeño', suggesting a possible origin for S(21)* pistil dysfunction. Two polymorphisms found within the S-RNase genomic region (a retrotransposon insertion within the intron of S(21)* and indels at the 3'UTR) might explain the different pattern of expression between S(21) and S(21)*. Evaluation of cultivars with unknown S-genotype identified another cultivar 'Azucar Verde' bearing S(21)*, and pollen tube growth examination confirmed self-compatibility for this cultivar as well. This is the first report of a mutated S-allele conferring self-compatibility in European pear.

  6. Study on the technology of the pear vinegar drink%雪梨醋饮的工艺研究

    Institute of Scientific and Technical Information of China (English)

    张智维; 张海群; 刘婷

    2014-01-01

    Through single factor experiment and orthogonal experiment ,using concentrated sydney juice as main material ,obtained the optimal process of alcoholic fermentation and ace-tic acid fermentation;then from the sydney vinegar ,Sydney juice ,honey and sucrose as raw material ,the optimum formulation of the pear vinegar drink are determined .The best process of alcohol fermentation is :amount of yeast 10% ,sugar 15% ,fermentation time 5 d .The opti-mum acetic acid fermentation is:amount of acetic acid bacteria 15% ,temperature of fermen-tation 28 ℃ ,The initial alcohol 6 .0% ,fermentation time 7 d .The best formula pear vinegar drink:pear vinegar 20% ,sucrose 4% ,pear juice 15% ,honey 6% .%通过单因素实验和正交试验设计,以浓缩雪梨汁为主要原料,得到了酒精发酵和醋酸发酵的最佳工艺;再以雪梨醋、雪梨汁、蜂蜜和蔗糖为原料,得到了雪梨醋饮的最佳配方.酒精发酵的最佳工艺为:酵母接种量10%,含糖量15%,发酵时间5 d .醋酸发酵的最佳工艺为:醋酸菌的接种量15%,发酵温度28℃,初始酒精度6.0%,发酵时间7 d .雪梨醋饮的最佳配方:雪梨醋20%,蔗糖4%,雪梨汁15%,蜂蜜6%.

  7. Improving the optimization efficiency and precision of least squares support vector regression (LSSVR) for pear property prediction

    Science.gov (United States)

    Hao, Yong; Liu, Yande; Zhang, Hailiang; Liu, Xuemei; Pan, Yuanyuan

    2010-10-01

    In this study, Visible/near-infrared (Vis/NIR) diffuse reflectance spectroscopy at 530-1560 nm region was investigated for the analysis of the soluble solids content (SSC) and color of pear. Least squares support vector regression (LSSVR) has been proven to be a powerful tool for modeling complex samples through the use of adapted kernel functions. However, one of the major drawbacks of LSSVR is that the optimization of the regularization and kernel meta-parameters is time-consuming during training the model, and the modeling results are sensitive to spectral noise. Wavelet compression pretreatment is an effective method for spectral information extraction and noise elimination. The calibration set was composed of 75 pear samples and 32 pear samples were used as the validation set. The raw and pretreated spectra by wavelet compression were modeled using LSSVR, It was shown that wavelet compression procedure not only shortened the modeling time, but also improved the predictive precision. The correlation coefficient (r) was improved from 0.78 to 0.93 for SSC, and from 0.95 to 0.96 for color, respectively. The root mean square error of prediction (RMSEP), optimization time and calibration variables were reduced from 0.68, 0.33s and 1031 to 0.41, 0.03s and 24 for SSC, while from 1.10, 0.33s and 1031 to 1.07, 0.03s and 40 for color. The results indicated that Vis/NIR spectroscopy combined with wavelet compression procedure and LSSVR is a reliable approach for predicting the SSC and color of pear.

  8. Determination of Quantities of Host Protein after Infection with Erwinia amylovora of Apple, Pear And Quince Cultivars

    Directory of Open Access Journals (Sweden)

    Şerife Çetin

    2014-10-01

    Full Text Available Fire blight disease caused by Erwinia amylovora is a destructive bacterial pathogen mainly on pears, apples and quinces from Rosaceae family. In this study, it was aimed determination of total protein amounts in different apple cultivars (Braeburn, Fuji, Gala and Golden, pear cultivars (Santa Maria and Williams and quince cultivars (Eşme and Ekmek in the infections of two virulent E. amylovora strains (Ea234-1 and Ea240-3 according as the time. It was taken leaf samples after leaf inoculation with E. amylovora (108 CFU ml-1 at 24th, 36th and 72nd hours. For verification of the infections, re-isolations were made from bacteria inoculated plants and the agent was identified as E. amylovora by biochemical, physiological and molecular tests. In determining the amounts of total protein and in the SDS-PAGE analyses were used Bradford and Laemmli methods, respectively, and absorbance values of protein extracts derived from the leaf samples taken, were obtained at 595 nm wavelength. According to the findings obtained; after infection of E. amylovora in the apple varieties comparing to controls, total protein concentrations at 24th hours increased and a decrease in the amount of 36th to 72nd hours and Braeburn has the highest protein content was determined. In the pear varieties, while total protein concentrations at 24th and 36th hours increased, a decrease in the amount of 72nd hour, and Santa Maria variety has the highest protein content was detected. In the quince varieties, total protein concentrations at 72th hour increased and Eşme variety has the highest protein content was identified. As a result of SDS-PAGE analysis, protein fractions which have different molecular weights were obtained. The protein bands were defined approximately 55-70 kDa and 35-55 kDa molecule weight on apple and quince varieties, respectively and also approx. 55-70 kDa in pear varieties.

  9. 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.

  10. Post-Fragmentation Whole Genome Amplification-Based Method

    Science.gov (United States)

    Benardini, James; LaDuc, Myron T.; Langmore, John

    2011-01-01

    This innovation is derived from a proprietary amplification scheme that is based upon random fragmentation of the genome into a series of short, overlapping templates. The resulting shorter DNA strands (genomic hybridization microarray, SNP analysis, and sequencing. The standard reaction can be performed with minimal hands-on time, and can produce amplified DNA in as little as three hours. Post-fragmentation whole genome amplification-based technology provides a robust and accurate method of amplifying femtogram levels of starting material into microgram yields with no detectable allele bias. The amplified DNA also facilitates the preservation of samples (spacecraft samples) by amplifying scarce amounts of template DNA into microgram concentrations in just a few hours. Based on further optimization of this technology, this could be a feasible technology to use in sample preservation for potential future sample return missions. The research and technology development described here can be pivotal in dealing with backward/forward biological contamination from planetary missions. Such efforts rely heavily on an increasing understanding of the burden and diversity of microorganisms present on spacecraft surfaces throughout assembly and testing. The development and implementation of these technologies could significantly improve the comprehensiveness and resolving power of spacecraft-associated microbial population censuses, and are important to the continued evolution and advancement of planetary protection capabilities. Current molecular procedures for assaying spacecraft-associated microbial burden and diversity have inherent sample loss issues at practically every step, particularly nucleic acid extraction. In engineering a molecular means of amplifying nucleic acids directly from single cells in their native state within the sample matrix, this innovation has circumvented entirely the need for DNA extraction regimes in the sample processing scheme.

  11. Evaluation of yeasts from Tibetan fermented products as agents for biocontrol of blue mold of Nashi pear fruits.

    Science.gov (United States)

    Hu, Hao; Xu, Yang; Lu, Huang-ping; Xiao, Rui; Zheng, Xiao-dong; Yu, Ting

    2015-04-01

    A total of 20 strains of yeast isolated from Tibetan fermented products were screened for antagonism against blue mold of pear caused by Penicillium expansum. Six isolates that inhibited incidence of postharvest decay by 35% or more were selected for further screening. Among them, the most effective was Rhodotorula mucilaginosa. The results showed that washed cell suspensions of R. mucilaginosa yielded better antagonistic efficacy than unwashed cell-culture mixtures, cell-free culture filtrates, and autoclaved cell cultures. Biocontrol activity improved with increasing concentrations of incubated cells. The best concentration was 1×10(8) cells/ml, at which the incidence of decay was only 16.7% after 6 d of incubation. The germination of conidia of P. expansum in vitro was significantly inhibited by both washed cell-suspensions and unwashed cell-culture mixtures. Rapid colonization by yeast at different concentrations showed a relationship between yeast-cell concentration and biocontrol activity. Although the titratable acidity of pear fruits increased after treatment, R. mucilaginosa did not affect the total soluble solids or ascorbic acid content. This is the first study to report that the yeast R. mucilaginosa from Tibet Autonomous Region of China may have potential as an antagonist to control the postharvest decay of pear fruits.

  12. 参梨活力饮工艺的研究%Participation Process of Pear Energy Drinks

    Institute of Scientific and Technical Information of China (English)

    刘文辉; 张炳盛; 王金凤; 陈莉; 刘娟娟

    2011-01-01

    采用西洋参提取液和莱阳梨汁作为原料,生产一种新型功能饮料,研究结果表明,两者合用能够补气养阴、润肺生津、化痰止咳、清火利咽,对于改善机体免疫力、增强体质有较好的辅助治疗作用。参梨活力饮的最佳配方为,西洋参须根1 kg、茎叶2 kg、莱阳梨汁192 kg,制备量966 L。%Study the use of American ginseng extract and Laiyang pear juice as raw material to produce a new functional beverage.Participation pear recipe for the best energy drink,1 kg American ginseng fibrous roots,stems and leaves of 2 kg,Laiyang pear juice 192kg

  13. The Ratio between Leave and Fruit Parameters on ‘William’ Pear Orchard Affected by Regulated Deficit Irrigation and Mulching

    Directory of Open Access Journals (Sweden)

    LAVDIM LEPAJA

    2016-03-01

    Full Text Available This field experiment was designed to assess the ratio between leave and fruit parameters on young ‘William’ pear trees after applied regulated deficit irrigation (RDI and mulching. Experiments related to deficit irrigation and particularly regulated deficit irrigation (RDI or partial rootzone drying depend heavily on weather conditions. Using a water budget methodology, four levels of irrigation, specifically 100% of evapotranspiration (ET as control and deficits of 80%, 60% and 40%, were applied to 10 trees during the season, 5 of which were mulched with wood chips at a 10 cm layer in first year of experiment while, 20 cm in second year. The experiment was conducted in Kosovo during 2013-2015 on a pear orchard of 10 ha using a nested experimental design. Using two-way ANOVA we found significant changes in a series of leave and fruit parameters. Our results confirmed that a moderate water stress increase yield while, reducing excessive vegetative growth. Regulated deficit irrigation (40 % has contributed to the reduction on leaf surface, leaf area, LAI. In addition, RDI affected to increase fruit numbers but decreasing fruit size. Compared with first year of experiment during 2015 in treatment 40 % were achieved 5 kg more than 2013 year. Except this, mulching had a positive effect on all parameter values measured compared to non-mulched trees. Our result indicated that regulated deficit irrigation can be successfully applied to pear also, RDI is an ideal water saving technique.

  14. 梨乳酸菌饮料的研制%Study on Pear Lactic Acid Bacteria Beverage

    Institute of Scientific and Technical Information of China (English)

    张旭光; 刘春芬; 慕金超

    2015-01-01

    以砀山梨汁和奶粉为原料,杀菌后接种乳酸菌混合发酵剂进行乳酸发酵,成为凝固型酸奶,进而调配成活性乳酸菌饮料。通过单因素和正交试验,得出梨乳酸菌饮料的优化条件:梨汁添加量为20%,加糖量10%,酸味剂用量0.15%,稳定剂用量0.4%。%The health care lactic acid bacteria beverage was fermented with lactobacilli by the raw materials of Dangshan pear juice and milk powder. The sterilization after inoculated lactic acid fermentation lactic acid bacteria culture blends.By single factor and orthogonal test , and the optimized conditions for pear lactic acid bacteria beverage:pear juice content is 20%, sugar 10%, citric acid 0.15%dosage, dosage of stabilizer 0.4%.

  15. Increasing water stress negatively affects pear fruit growth by reducing first its xylem and then its phloem inflow.

    Science.gov (United States)

    Morandi, Brunella; Losciale, Pasquale; Manfrini, Luigi; Zibordi, Marco; Anconelli, Stefano; Galli, Fabio; Pierpaoli, Emanuele; Corelli Grappadelli, Luca

    2014-10-15

    Drought stress negatively affects many physiological parameters and determines lower yields and fruit size. This paper investigates on the effects of prolonged water restriction on leaf gas exchanges, water relations and fruit growth on a 24-h time-scale in order to understand how different physiological processes interact to each other to face increasing drought stress and affect pear productive performances during the season. The diurnal patterns of tree water relations, leaf gas exchanges, fruit growth, fruit vascular and transpiration flows were monitored at about 50, 95 and 145 days after full bloom (DAFB) on pear trees of the cv. Abbé Fétel, subjected to two irrigation regimes, corresponding to a water restitution of 100% and 25% of the estimated Etc, respectively. Drought stress progressively increased during the season due to lower soil tensions and higher daily vapour pressure deficits (VPDs). Stem water potential was the first parameter to be negatively affected by stress and determined the simultaneous reduction of fruit xylem flow, which at 95 DAFB was reflected by a decrease in fruit daily growth. Leaf photosynthesis was reduced only from 95 DAFB on, but was not immediately reflected by a decrease in fruit phloem flow, which instead was reduced only at 145 DAFB. This work shows how water stress negatively affects pear fruit growth by reducing first its xylem and then its phloem inflow. This determines a progressive increase in the phloem relative contribution to growth, which lead to the typical higher dry matter percentages of stressed fruit.

  16. 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.

  17. 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.

  18. Detection and Sequence Analysis of the Apple Scar Skin Viroid Isolated from Pear Trees in Xinjiang%新疆梨树上苹果锈果类病毒的检测与全序列分析

    Institute of Scientific and Technical Information of China (English)

    王钰婷; 金珠; 董芳园; 和世玉; 张莉; 牛建新

    2013-01-01

    [目的]检测并分析新疆和静县223团场多年生早熟梨树上的苹果锈果类病毒(ASSVd).[方法]对采自新疆和静县223团多年生早熟梨的枝条和叶片样品进行RNA抽提,经RT-PCR技术鉴定检测.[结果](1)多年生早熟梨树检测到苹果锈果类病毒,得到2条ASSVd核酸序列(JX861258~JX861259),所得序列与G enBank中已报道的国内外ASSVd序列同源性达86%以上.(2)原位RT-PCR检测进一步表明ASSVd的RNA阳性信号主要位于叶片叶肉组织的细胞核内.[结论]通过序列分析比对,各寄主上的ASSVd分离物核酸序列变异较小,地域和品种间核酸序列无明显差异.建立了优化的RT-PCR检测及原位RT-PCR检测方法,为果树ASSVd的快速检测奠定了良好的基础.%[ Objective and Method] In order to detect and analyze the ASSVd in precocious pear trees, low molecular weight RNAs were extracted from tender leaves and shoots of precocious pear trees which were collected from No. 223 Farming & Herding Regiment in Hejing County, Xinjiang Uygur Autonomous Region. Then the samples were detected by reverse transcription - polymerase chain reaction ( RT - PCR). [ Result] The results showed; (1 )Two of the precocious pear trees were infected with Apple scar skin viroid (ASSVd) and the 2 isolates (accession numbers JX861258 - JX861259) had over 86% nucleotide sequence identity with previously published sequences in the GenBank. (2) In situ RT - PCR further confirmed the existence of ASSVd in the leaf tissues and indicated that ASSVd was mainly distributed in the nucleus. [ Conclusion ] There was no geographic and variety differentiation between ASSVd isolates. Optimized detection methods of RT - PCR and In situ RT - PCR were established, thus providing the basis for the rapid identification of ASSVd in fruit trees.

  19. Development of PCR internal controls for DNA profiling with the AmpFℓSTR® SGM Plus® amplification kit.

    Science.gov (United States)

    Nathalie, Zahra; Hadi, Sibte; Goodwin, William

    2012-09-01

    Forensic DNA profiling uses a series of commercial kits that co-amplify several loci in one reaction; the products of the PCR are fluorescently labelled and analysed using CE. Before CE, an aliquot of the PCR is mixed with formamide and an internal lane size standard. Using the SGM Plus amplification kit, we have developed two internal non-amplified controls of 80 bp and 380 bp that are labelled with ROX fluorescent dye and added to the PCR. Combined with two internal amplification controls of 90 bp and 410 bp, they provide additional controls for the PCR, electrokinetic injection, and CE and also function as an internal size standard.

  20. 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

  1. Preparation of Health Drink of Balsam Pear and Apple%苦瓜苹果保健饮料的研制

    Institute of Scientific and Technical Information of China (English)

    黄运凤; 刘国凌

    2009-01-01

    [Objective] The study was to determine the optimal technical formula for health drink of balsam pear and apple. [ Method] With balsam pear and apple as the main materials, the health drink of balsam pear and apple was prepared. The effects of amounts of balsam pear juice, apple juice, honey and citric acid on quality of the product were studied through orthogonal test. [ Result ] The effects of main factors on quality of the product from big to small in order was balsam pear juice amount > citric acid amount > honey amount > apple juice amount. The optimal technical formula for health drink of balsam pear and apple was 20% balsam pear juice + 10% apple juice + 1. 5 g honey +0. 035 g citric acid. [ Conclusion ] Under the optimal technological conditions, the product was refreshing and sweet with rich nutrition and suitable taste.%[目的]确定苦瓜苹果保健饮料的最佳工艺配方.[方法]以苦瓜和苹果为主要原料制备苦瓜苹果保健饮料,采用正交试验研究苦瓜汁、苹果汁、蜂蜜和柠檬酸用量对产品品质的影响.[结果]影响产品品质的主要因素由大到小依次为:苦瓜汁用量>柠檬酸用量>蜂蜜用量>苹果汁用量;苦瓜苹果保健饮料的最佳工艺配方为:20%苦瓜汁+10%苹果汁+1.5 g蜂蜜+0.035 g柠檬酸.[结论]在最佳工艺条件下所制混合饮料清新甘甜、营养丰富、口感适宜.

  2. A cascade signal amplification strategy for surface enhanced Raman spectroscopy detection of thrombin based on DNAzyme assistant DNA recycling and rolling circle amplification.

    Science.gov (United States)

    Gao, Fenglei; Du, Lili; Tang, Daoquan; Lu, Yao; Zhang, Yanzhuo; Zhang, Lixian

    2015-04-15

    A sensitive protocol for surface enhanced Raman spectroscopy (SERS) detection of thrombin is designed with R6G-Ag NPs as a signal tag by combining DNAzyme assistant DNA recycling and rolling circle amplification (RCA). Molecular beacon (MB) as recognition probe immobilizes on the glass slides and performs the amplification procedure. After thrombin-induced structure-switching DNA hairpins of probe 1, the DNAzyme is liberated from the caged structure, which hybridizes with the MB for cleavage of the MB in the presence of cofactor Zn(2+) and initiates the DNA recycling process, leading to the cleavage of a large number of MB and the generation of numerous primers for triggering RCA reaction. The long amplified RCA product which contained hundreds of tandem-repeat sequences, which can bind with oligonucleotide functionalized Ag NPs reporters. The attached signal tags can be easily read out by SERS. Because of the cascade signal amplification, these newly designed protocols provides a sensitive SERS detection of thrombin down to the femolar level (2.3fM) with a linear range of 5 orders of magnitude (from 10(-14) to 10(-9)M) and have high selectivity toward its target protein. The proposed method is expected to be a good clinical tool for the diagnosis of a thrombotic disease.

  3. Study on the mating compatibility of part pear varieties and wild types of Pyrus ussuriensis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    To understand the mating compatibility of Pyrus ussuriensis Maxim.,we studied the fertility of pollen and conducted a hand-pollination trial in the field on some pear varieties and wild types.The results showed that about 53% of varieties among 32 tested genotypes were male sterile.Not only did the pollen vitalities in normal varieties show distinct differences,but pollen vitalities from flower forcing in a glasshouse were found to be lower than those from natural flowering in the field,which had no apparent effect on fruit setting of tested varieties.Most of the tested genotypes such as Nanguoli,Pingxiangli,and Hanxiangli showed selfincompatibility (SI).Honghuagaili could bear fruit after hand pollination,but there were abnormal seeds in its fruits.So we suggested it was a recessive SI that happened during embryo development.Longxiangli has the capacity of self-compatibility (SC) to some extent,its fruit setting rate of inflorescence could reach 23.3%.Manual self-pollination during bud flowering could improve the fruit setting rate of part tested genotypes with SI,but had no effect on the fruit setting rate 3 days after flowering.Mating between female parents with the variety selected from F1 generation showed that the majority of their combinations were compatible.There was one-way SC when Nanguoli was crossed with Hanhongli,while no fruits could be found after Hanhongli was crossed with Nanguoli.It may be related to the S-genotype or haplotype of Nanguoli.In addition,mating between the varieties derived from bud mutation with the female parent appeared incompatible.We concluded that P.ussuriensis Maxim.is similar to other grown pear systems with the characteristics of SI,the fruit setting rate of self pollination in some varieties and wild types can be improved by artificial self-pollination during bud flowering,and fruit cannot be developed through pollination between the varieties from bud mutation and the female parent.

  4. 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.

  5. Development of a Loop-Mediated Isothermal Amplification Assay for Porcine Circovirus Type 2

    Institute of Scientific and Technical Information of China (English)

    Ye-bing Liu; Lei Zhang; Qin-hong Xue; Yi-bao Ning; Zhi-gang Zhang

    2011-01-01

    In this study,the loop-mediated isothermal amplification(LAMP)method was used to develop a rapid and simple detection system for porcine circovirus type 2(PCV2).According to the PCV2 sequences published in GenBank,multiple LAMP primers were designed targeting conserved sequences of PCV2.Using the DNA extracted from PCV2 isolates HUN-09 and SD-09 as the template,LAMP reactions in a PCV2 LAMP system was performed,the amplification products were detected by adding SYBR Green I and could be observed directly by the naked eye.The results showed highly-efficient and specific amplification in 30 min at 63℃ with a LAMP real-time turbidimeter.Furthermore,PCV2 DNA templates,with a detection limit of 5.5×10-5ng of nucleic acid,indicated that this assay was highly sensitive.The results obtained with the naked eye after SYBR Green I staining were consistent with those detected by the real-time turbidimeter,showing the potential simplicity of interpretation of the assay results.The LAMP assay appeared to have greater accuracy than PCR and virus isolation for the analysis of 18 clinical samples.In addition it offers higher specificity and sensitivity,shorter reaction times and simpler procedures than the currently available methods of PCV2 detection.It is therefore a promising tool for the effective and efficient detection of PCV2.

  6. A new subtype of high-grade mandibular osteosarcoma with RASAL1/MDM2 amplification.

    Science.gov (United States)

    Guérin, Maxime; Thariat, Juliette; Ouali, Mounia; Bouvier, Corinne; Decouvelaere, Anne-Valérie; Cassagnau, Elisabeth; Aubert, Sébastien; Lepreux, Sébastien; Coindre, Jean-Michel; Valmary-Degano, Séverine; Larousserie, Frédérique; Meilleroux, Julie; Projetti, Fabrice; Stock, Nathalie; Galant, Christine; Marie, Béatrice; Peyrottes, Isabelle; de Pinieux, Gonzague; Gomez-Brouchet, Anne

    2016-04-01

    In contrast to long bone osteosarcoma, mandibular osteosarcoma is highly heterogeneous and morphologically overlaps with benign tumors, obscuring diagnosis and treatment selection. Molecular characterization is difficult due to the paucity of available specimens of this rare disease. We aimed to characterize the spectrum of mandibular osteosarcoma using immunohistochemistry and molecular techniques (quantitative polymerase chain reaction and sequencing) and compare them with benign fibro-osseous lesions. Forty-nine paraffin-embedded mandible osteosarcoma tissue samples were collected retrospectively and compared with 10 fibrous dysplasia and 15 ossifying fibroma cases. These were analyzed for molecular markers thought to differ between the different diseases and subtypes: MDM2 (murine double-minute type 2) overexpression, GNAS (guanine nucleotide-binding protein/α subunit) mutations, and amplification of MDM2 and/or RASAL1 (RAS protein activator like 1). Five fibroblastic high-grade osteosarcoma subtypes showed MDM2 amplification, including 2 with a microscopic appearance of high-grade osteosarcoma with part low-grade osteosarcoma (differentiated/dedifferentiated osteosarcoma) and MDM2 overexpression. The other 3 contained a coamplification of MDM2 and RASAL1, a signature also described for juvenile ossifying fibroma, with no overexpression of MDM2. These were of the giant cell-rich high-grade osteosarcoma, with areas mimicking juvenile ossifying fibroma (ossifying fibroma-like osteosarcoma). Our results show that some diagnosed high-grade osteosarcomas are differentiated/dedifferentiated osteosarcomas and harbor an overexpression and amplification of MDM2. In addition, juvenile ossifying fibromas can potentially evolve into giant cell-rich high-grade osteosarcomas and are characterized by a RASAL1 amplification (osteosarcoma with juvenile ossifying fibroma-like genotype). Thus, the presence of a RASAL1 amplification in ossifying fibroma may indicate a requirement

  7. Amplification Without Inversion in Semiconductor Quantum Dot

    Science.gov (United States)

    Hajibadali, A.; Abbasian, K.; Rostami, A.

    In this paper, we have realized amplification without inversion (AWI) in quantum dot (QD). A Y-type four-level system of InxGa1-xN quantum dot has been obtained and investigated for AWI. It has been shown that, with proper setting of control fields' amplitude, we can obtain reasonable gain. With proper setting of phase difference of control fields and probe field, we can obtain considerable gain in resonant wavelength. We have designed this system by solving the Schrödinger-Poisson equations for InxGa1-xN quantum dot in GaN substrate, self-consistently.

  8. 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.

  9. Amplification and characterization of eukaryotic structural genes.

    Science.gov (United States)

    Maniatis, T; Efstratiadis, A; Sim, G K; Kafatos, F

    1978-05-01

    An approach to the study of eukaryotic structural genes which are differentially expressed during development is described. This approach involves the isolation and amplification of mRNA sequences by in vitro conversion of mRNA to double-stranded cDNA followed by molecular cloning in bacterial plasmids. This procedure provides highly specific hybridization probes that can be used to identify genes and their contiguous DNA sequences in genomic DNA, and to detect specific RNA transcripts during development. The nature of the method allows the isolation of individual mRNA sequences from a complex population of molecules at different stages of development.

  10. A novel whole genome amplification method using type IIS restriction enzymes to create overhangs with random sequences.

    Science.gov (United States)

    Pan, Xiaoming; Wan, Baihui; Li, Chunchuan; Liu, Yu; Wang, Jing; Mou, Haijin; Liang, Xingguo

    2014-08-20

    Ligation-mediated polymerase chain reaction (LM-PCR) is a whole genome amplification (WGA) method, for which genomic DNA is cleaved into numerous fragments and then all of the fragments are amplified by PCR after attaching a universal end sequence. However, the self-ligation of these fragments could happen and may cause biased amplification and restriction of its application. To decrease the self-ligation probability, here we use type IIS restriction enzymes to digest genomic DNA into fragments with 4-5nt long overhangs with random sequences. After ligation to an adapter with random end sequences to above fragments, PCR is carried out and almost all present DNA sequences are amplified. In this study, whole genome of Vibrio parahaemolyticus was amplified and the amplification efficiency was evaluated by quantitative PCR. The results suggested that our approach could provide sufficient genomic DNA with good quality to meet requirements of various genetic analyses.

  11. Cascaded multiple amplification strategy for ultrasensitive detection of HIV/HCV virus DNA.

    Science.gov (United States)

    Wang, Kun; Fan, Daoqing; Liu, Yaqing; Dong, Shaojun

    2017-01-15

    Ultrasensitive detection of HIV and HCV virus DNA is of great importance for early accurate diagnostics and therapy of HIV virus-infected patients. Herein, to our best knowledge, it is the first to use DNA cascaded multiple amplification strategy for ultrasensitive detection of HIV virus DNA with G-quadruplex-specific fluorescent or colorimetric probes as signal carriers. The developed strategy also exhibited universal applicability for HCV virus DNA detection. After reaction for about 4h, high sensitivity and specificity can be achieved at both fluorescent and colorimetric strategies (limit of detection (LOD) of 10 fM and 0.5pM were reached for fluorescent and colorimetric detection, respectively). And the single-based mismatched DNA even can be distinguished by naked eyes. It is believed that the cascaded multiple amplification strategy presents a huge advance in sensing platform and potential application in future clinical diagnosis.

  12. ANALYSIS OF C-HA-RAS GENE AMPLIFICATION AND MUTATION IN LARYNGEAL CARCINOMA

    Institute of Scientific and Technical Information of China (English)

    刘世喜; 林代诚; 洪邦泰; 黄光琦

    1995-01-01

    In order to study the ahered molecular events during laryngeal carcinogenesis and elucidate the role of Ha-ras oncogene amplification and mutation, we have examined their profile by polymerase chain reaction (PCR) and selective oligonucleoride hybridization. We analyzed the mutational status of codon 12 of Ha-ras in 22 laryngeal carcinomas and 10 normal tissues, and found that 7 of 22 laryngeal carcinomas con-tained a Ha-ras mutation at codon 12. The frequency of mutation was 32%. None of the normal tissues re-vealed mutation. Moreover, no amplification was found in cancers when compared to the normal. Our findings indicated that the aefivmed Ha-ras gene existed in laryngeal carcinoma, and activation of the Ha-ras gene by mutation at codon 12 might play a key role in laryngeal carcinogenesis.

  13. A simple molecular beacon with duplex-specific nuclease amplification for detection of microRNA.

    Science.gov (United States)

    Li, Yingcun; Zhang, Jiangyan; Zhao, Jingjing; Zhao, Likun; Cheng, Yongqiang; Li, Zhengping

    2016-02-01

    MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene activity, promoting or inhibiting cell proliferation, migration and apoptosis. Abnormal expression of miRNAs is associated with many diseases. Therefore, it is essential to establish a simple, rapid and sensitive miRNA detection method. In this paper, based on a simple molecular beacon (MB) and duplex-specific nuclease (DSN), we developed a target recycling amplification method for miRNA detection. By controlling the number of stem bases to 5, the MB probe used in this method can be prevented from hydrolysis by DSN without special modification. This assay is direct and simple to quantitatively detect miRNA with high sensitivity and specificity. The MB probe design provides a new strategy for nuclease-based amplification reaction.

  14. Rapid amplification of genetically modified organisms using a circular ferrofluid-driven PCR microchip.

    Science.gov (United States)

    Sun, Yi; Kwok, Yien-Chian; Foo-Peng Lee, Peter; Nguyen, Nam-Trung

    2009-07-01

    The use of genetically modified organisms (GMOs) as food and in food products is becoming more and more widespread. Polymerase chain reaction (PCR) technology is extensively used for the detection of GMOs in food products in order to verify compliance with labeling requirements. In this paper, we present a novel close-loop ferrofluid-driven PCR microchip for rapid amplification of GMOs. The microchip was fabricated in polymethyl methacrylate by CO2 laser ablation and was integrated with three temperature zones. PCR solution was contained in a circular closed microchannel and was driven by magnetic force generated by an external magnet through a small oil-based ferrofluid plug. Successful amplification of genetically modified soya and maize were achieved in less than 13 min. This PCR microchip combines advantages of cycling flexibility and quick temperature transitions associated with two existing microchip PCR techniques, and it provides a cost saving and less time-consuming way to conduct preliminary screening of GMOs.

  15. Mechanism of seasonal Arctic sea ice evolution and Arctic amplification

    OpenAIRE

    Kim, Kwang-Yul; Hamlington, Benjamin D.; Na, Hanna; Kim, Jinju

    2016-01-01

    Sea ice loss is proposed as a primary reason for the Arctic amplification, although the physical mechanism of the Arctic amplification and its connection with sea ice melting is still in debate. In the present study, monthly ERA-Interim reanalysis data are analyzed via cyclostationary empirical orthogonal function analysis to understand the seasonal mechanism of sea ice loss in the Arctic Ocean and the Arctic amplification. While sea ice loss is widespread over much of the p...

  16. Microscale Mechanism of Age Dependent Wetting Properties of Prickly Pear Cacti (Opuntia).

    Science.gov (United States)

    Rykaczewski, Konrad; Jordan, Jacob S; Linder, Rubin; Woods, Erik T; Sun, Xiaoda; Kemme, Nicholas; Manning, Kenneth C; Cherry, Brian R; Yarger, Jeffery L; Majure, Lucas C

    2016-09-13

    Cacti thrive in xeric environments through specialized water storage and collection tactics such as a shallow, widespread root system that maximizes rainwater absorption and spines adapted for fog droplet collection. However, in many cacti, the epidermis, not the spines, dominates the exterior surface area. Yet, little attention has been dedicated to studying interactions of the cactus epidermis with water drops. Surprisingly, the epidermis of plants in the genus Opuntia, also known as prickly pear cacti, has water-repelling characteristics. In this work, we report that surface properties of cladodes of 25 taxa of Opuntia grown in an arid Sonoran climate switch from water-repelling to superwetting under water impact over the span of a single season. We show that the old cladode surfaces are not superhydrophilic, but have nearly vanishing receding contact angle. We study water drop interactions with, as well as nano/microscale topology and chemistry of, the new and old cladodes of two Opuntia species and use this information to uncover the microscopic mechanism underlying this phenomenon. We demonstrate that composition of extracted wax and its contact angle do not change significantly with time. Instead, we show that the reported age dependent wetting behavior primarily stems from pinning of the receding contact line along multilayer surface microcracks in the epicuticular wax that expose the underlying highly hydrophilic layers.

  17. 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.

  18. Temporal Dependency of Yield and Quality Estimation through Spectral Vegetation Indices in Pear Orchards

    Directory of Open Access Journals (Sweden)

    Jonathan Van Beek

    2015-08-01

    Full Text Available Yield and quality estimations provide vital information to fruit growers, yet require accurate monitoring throughout the growing season. To this end, the temporal dependency of fruit yield and quality estimations through spectral vegetation indices was investigated in irrigated and rainfed pear orchards. Both orchards were monitored throughout three consecutive growing seasons, including spectral measurements (i.e., hyperspectral canopy reflectance measurements as well as yield determination (i.e., total yield and number of fruits per tree and quality assessment (i.e., fruit firmness, total soluble solids and fruit color. The results illustrated a clear association between spectral vegetation indices and both fruit yield and fruit quality (|r| > 0.75; p < 0.001. However, the correlations between vegetation indices and production variables varied throughout the growing season, depending on the phenological stage of fruit development. In the irrigated orchard, index values showed a strong association with production variables near time of harvest (|r| > 0.6; p < 0.001, while in the rainfed orchard, index values acquired during vegetative growth periods presented stronger correlations with fruit parameters (|r| > 0.6; p < 0.001. The improved planning of remote sensing missions during (rainfed orchards and after (irrigated orchards vegetative growth periods could enable growers to more accurately predict production outcomes and improve the production process.

  19. 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.

  20. 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.

  1. 红梨新品种'红月梨'%A New Red Pear Cultivar'Hongyue'

    Institute of Scientific and Technical Information of China (English)

    李俊才; 刘成; 王家珍; 蔡忠民; 沙守峰

    2011-01-01

    '红月梨'是由'红茄梨'和'苹果梨'杂交育成.果实圆锥形,平均单果质量245 g.表面红色,果皮薄.果肉白色,后熟后肉质细腻多汁,风味酸甜,微香,石细胞少,含可溶性固形物14.4%,总糖10.33%,可滴定酸0.34%,维生素C 0.0248 mg·g-1,品质优.在辽宁熊岳地区9月上旬成熟,早产,旱丰,优质,易栽培管理,抗病,抗寒.%‘Hongyue’ pear is bred by the cross ‘Red Clapp's’ × ‘Apple-pear’ . Fruit is conical and single fruit weigh is 245 g with red skin. Fruit skin is thin. Flesh is white and juicy after ripening. Flavor is sour-sweet with a little aroma, and less stone cell. The soluble solids content is 14.4%, total sugar conternt is 10.33%, titratable acid content is 0.34%, vitamin C content is 0.0248 mg· g-1. Fruit quality is high. It ripens in early September in Xiongyue, Liaoning. It is a variety with high and early productive, high quality, resistance to diseases and chilling.

  2. Free amino acids in the xylem sap of pear trees during dormancy

    Directory of Open Access Journals (Sweden)

    Anderson Carlos Marafon

    2016-01-01

    Full Text Available ABSTRACT: Storage and remobilization are considered key processes for the effective use of nitrogen in temperate fruit trees. As dormancy begins, storage proteins are synthesized, coinciding with a reduction in the levels of free amino acids. Consequently, as dormancy breaks, these storage proteins are degraded, and an increase in the concentrations of amino acids occurs, in order to support new growth. The objective of this study was to evaluate water content of different vegetative tissues (buds, bark, and bole wood, volume of xylem sap, and free amino acid concentrations of xylem sap, during winter dormancy of Hosui Japanese pear trees (VL. Plant material was obtained from the Embrapa Temperate Climate experimental orchard at Pelotas, in the state of Rio Grande do Sul, Brazil. Xylem sap was extracted from the branches with the aid of a vacuum pump, and the free amino acids were determined by gas chromatography, using the EZ kit: Faast GC/FID (Phenomenex. Water content of buds, as well as the volume of sap and concentrations of both aspartic acid and asparagine, substantially increased over time, reaching maximum values in the phase preceding sprouting.

  3. Yield and vegetative growth of cactus pear at different spacings and under chemical fertilizations

    Directory of Open Access Journals (Sweden)

    João A. da Silva

    2016-06-01

    Full Text Available ABSTRACT The objective was to evaluate the effect of different spacings and mineral fertilizations on cactus pear growth and production in a randomized block design, with three replicates, in a 3 x 4 factorial scheme: three spacings, 1.00 x 0.50 m, 2.00 x 0.25 m and 3.00 x 1.00 x 0.25 m, and four fertilizations, 000-000-000, 000-150-000, 200-150-000 and 200-150-100 kg ha-1 of N, P2O5 and K2O, respectively. Plant growth was evaluated between 90 and 390 days and production and growth were evaluated at 620 days after planting. There were significant interactions between spacing and fertilization for plant height, number of cladodes and cladode area index from 90 to 390 days and for production of fresh and dry matter at 620 days after planting. Spacing influenced cladode area index, while fertilization influenced plant height, number of cladodes and cladode area index at 620 days after planting. Plant height showed cubic effect for the days after planting. Number of cladodes and cladode area index were dependent on spacing, fertilization and plant age, and fitted to cubic models. The best results of growth and production of fresh and dry matter are associated with NPK and NP fertilizations and the spacing of 1.00 x 0.50 m.

  4. Where does Grapholita molesta (Busck) (Lepidoptera: Tortricidae) overwinter in adjacent peach, pear and apple orchards?

    Science.gov (United States)

    Yang, X-F; Fan, F; Wang, C; Wei, G-S

    2016-02-01

    The Oriental fruit moth, Grapholita molesta (Lepidoptera: Tortricidae), is a major pest of tree fruits worldwide, and the diapausing larvae overwinter in cryptic habitats. Investigations of overwintering G. molesta were conducted in adjacent peach, pear and apple orchards in Northern China over three consecutive winters to determine the overwintering site and habitat preferences of the moth. Counts of overwintering larvae in the different orchards demonstrated that the late-maturing peach orchard ('Shenzhou honey peach') was the most preferred overwintering habitat with more than 90% of the collected larvae. Larvae were more abundant in host trees, and they very rarely overwintered in the soil. The overwintering site preferences on the host trees were significantly different; over 50% larvae were located in the tree trunks, and followed by main branches. Most of the G. molesta overwintered on the sunny side of the host trees at or below 60 cm from the ground; a few were cocooned on the shaded sides of the trees or greater than 60 cm from the ground. G. molesta began overwintering between August and October, mid- to late September was the peak period for entering winter diapause during 2011-2013 (77.78, 67.59 and 71.15%, respectively). Our findings improve understanding of the orchard habitat and overwintering site preferences of G. molesta and would be useful in the development of efficient forecasting and pest-management strategies for orchards during the winter and early spring.

  5. 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.

  6. Erwinia uzenensis sp. nov., a novel pathogen that affects European pear trees (Pyrus communis L.).

    Science.gov (United States)

    Matsuura, Takayuki; Mizuno, Akifumi; Tsukamoto, Takanori; Shimizu, Yoshiaki; Saito, Norihiko; Sato, Shigeyoshi; Kikuchi, Shigemi; Uzuki, Tsuneyasu; Azegami, Koji; Sawada, Hiroyuki

    2012-08-01

    Bacteria were isolated from black lesions on shoots of European pear trees (Pyrus communis L.) in an orchard in Japan. Previous characterization of this novel pathogen by phenotypic and genotypic methods suggested that it should belong to the genus Erwinia but might not correspond to either Erwinia amylovora or Erwinia pyrifoliae. Here, phylogenetic analyses of the 16S rRNA gene, gyrB, and rpoD gene sequences indicated that it could not be assigned to any recognized species of the genus Erwinia. DNA-DNA hybridization confirmed that the bacterial strains represented a novel species. The DNA G+C contents, the fatty acid profile and phenotypic characteristics resembled those previously reported for members of the genus Erwinia. On the basis of these and previous results, the pathogen represents a novel species of the genus Erwinia, for which the name Erwinia uzenensis sp. nov. (type strain: YPPS 951(T) = LMG 25843(T) = NCPPB 4475(T)) is proposed.

  7. Identification of Erwinia species isolated from apples and pears by differential PCR.

    Science.gov (United States)

    Gehring, I; Geider, K

    2012-04-01

    Many pathogenic and epiphytic bacteria isolated from apples and pears belong to the genus Erwinia; these include the species E. amylovora, E. pyrifoliae, E. billingiae, E. persicina, E. rhapontici and E. tasmaniensis. Identification and classification of freshly isolated bacterial species often requires tedious taxonomic procedures. To facilitate routine identification of Erwinia species, we have developed a PCR method based on species-specific oligonucleotides (SSOs) from the sequences of the housekeeping genes recA and gpd. Using species-specific primers that we report here, differentiation was done with conventional PCR (cPCR) and quantitative PCR (qPCR) applying two consecutive primer annealing temperatures. The specificity of the primers depends on terminal Single Nucleotide Polymorphisms (SNPs) that are characteristic for the target species. These PCR assays enabled us to distinguish eight Erwinia species, as well as to identify new Erwinia isolates from plant surfaces. When performed with mixed bacterial cultures, they only detected a single target species. This method is a novel approach to classify strains within the genus Erwinia by PCR and it can be used to confirm other diagnostic data, especially when specific PCR detection methods are not already available. The method may be applied to classify species within other bacterial genera.

  8. Developmental changes in composition and quality of prickly pear cactus cladodes (nopalitos).

    Science.gov (United States)

    Rodriguez-Felix, A; Cantwell, M

    1988-01-01

    The composition and quality of edible tender stems or cladodes of 3 Prickly Pear Cactus species (Opuntia amyclaea, O. ficus-indica, and O. inermis) were studied at different stages of development. This traditional Mexican vegetable is called "nopalitos" in Spanish and "cactus leaves" in English. Cladodes harvested when 20 cm in length have the following average composition per 100 g: 91.7 g of water, 1.1 g of protein, 0.2 g of lipid, 1.3 g of ash, 1.1 g of crude fiber, 4.6 g of complex carbohydrates and 0.82 g of simple sugars, 12.7 mg of ascorbic acid and 28.9 micrograms of carotenes. The cladode's juice has an average pH of 4.6, 0.45% titratable acidity and 6.9% soluble solids. The components which varied most during development of the cladodes were: carotenes, acidity and total carbohydrates which increased, and protein and crude fiber (acid-detergent) which decreased. The nutritive value of the tender cladodes in the stages of growth at which they are commonly harvested and consumed (15 to 25 cm long weighing 50 to 80 g per stem), was similar for the 3 species.

  9. Cactus pear cladodes powders as a source of dietary fibre: purification and properties.

    Science.gov (United States)

    Saenz, Carmen; Yoong, Maylin; Figuerola, Fernando; Chiffelle, Italo; María Estevez, Ana

    2012-05-01

    Cactus pear cladodes of 2-3 years were used to obtain a natural purified dietary fibre and their physical, chemical and technological properties were determined. The effect of particle size and washing temperature on the technological properties was studied. Purification produces a decrease in green colour (a*) and an increase in total dietary fibre but reduces the total phenolic compounds, mainly when cladodes are washed at higher temperatures. Technological properties did not present changes in the water retention capacity (WRC), water adsorption capacity and cationic exchange capacity, but it did in swelling capacity (SC), oil absorption capacity, apparent density and setting density, which were influenced by the particle size of the cactus powders. The purified fibre shows a high WRC between 5.20 and 5.86 g g(- 1) and a high SC (7.02-8.27 mL g(- 1)). Purified fibre with a particle size between 600 and 1200 μm, independent of the washing temperature had better insoluble to soluble dietary fibre ratio, total phenolic content and technological properties.

  10. 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.

  11. Kaposi肉瘤中人疱疹病毒8的检测及其临床病理学意义%Detection of human herpes virus 8 in Kaposi sarcoma using polymerase chain reaction and single stranded probe in situ hybridization with a tyramide signal amplification system

    Institute of Scientific and Technical Information of China (English)

    周小鸽; 张小平; 等

    2002-01-01

    目的了解Kaposi肉瘤与人疱疹病毒8(HHV8)或Kaposi肉瘤相关病毒的关系;建立适合于临床病理诊断的原位杂交方法.方法采用地高辛标记的单链HHV8 DNA探针,辅以增强探测敏感性的信号扩增系统(tyramide signal amplification)对14例甲醛固定石蜡包埋的Kaposi肉瘤组织标本进行了HHV8原位检测,为了比较,同时也采用了聚合酶链反应(PCR)检测技术.结果用原位杂交和PCR两种方法共发现10例(71%)Kaposi肉瘤组织中存在HHV8感染.其中8例在两种技术检测中均呈阳性. HHV8阳性信号见于肿瘤梭形细胞和内皮细胞,也见于各期病变(早中期的斑片和斑块病变,晚期的结节病变),以及原发,复发和(或)转移病变.所有阴性对照在两种方法中均呈阴性.结论 Kaposi肉瘤与HHV8存在密切关系;这种地高辛标记单链DNA探针,辅以信号扩增系统的原位杂交技术检测该病毒具有用于临床病理诊断的价值.

  12. 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.

  13. Magnetic Field Amplification in Young Galaxies

    CERN Document Server

    Schober, Jennifer; Klessen, Ralf S

    2013-01-01

    The Universe at present is highly magnetized, with fields of the order of a few 10^-5 G and coherence lengths larger than 10 kpc in typical galaxies like the Milky Way. We propose that the magnetic field was amplified to this values already during the formation and the early evolution of the galaxies. Turbulence in young galaxies is driven by accretion as well as by supernova (SN) explosions of the first generation of stars. The small-scale dynamo can convert the turbulent kinetic energy into magnetic energy and amplify very weak primordial magnetic seed fields on short timescales. The amplification takes place in two phases: in the kinematic phase the magnetic field grows exponentially, with the largest growth on the smallest non-resistive scale. In the following non-linear phase the magnetic energy is shifted towards larger scales until the dynamo saturates on the turbulent forcing scale. To describe the amplification of the magnetic field quantitatively we model the microphysics in the interstellar medium ...

  14. Experimental noiseless linear amplification using weak measurements

    Science.gov (United States)

    Ho, Joseph; Boston, Allen; Palsson, Matthew; Pryde, Geoff

    2016-09-01

    The viability of quantum communication schemes rely on sending quantum states of light over long distances. However, transmission loss can degrade the signal strength, adding noise. Heralded noiseless amplification of a quantum signal can provide a solution by enabling longer direct transmission distances and by enabling entanglement distillation. The central idea of heralded noiseless amplification—a conditional modification of the probability distribution over photon number of an optical quantum state—is suggestive of a parallel with weak measurement: in a weak measurement, learning partial information about an observable leads to a conditional back-action of a commensurate size. Here we experimentally investigate the application of weak, or variable-strength, measurements to the task of heralded amplification, by using a quantum logic gate to weakly couple a small single-optical-mode quantum state (the signal) to an ancilla photon (the meter). The weak measurement is carried out by choosing the measurement basis of the meter photon and, by conditioning on the meter outcomes, the signal is amplified. We characterise the gain of the amplifier as a function of the measurement strength, and use interferometric methods to show that the operation preserves the coherence of the signal.

  15. Seismic Wave Amplification in 3D Alluvial Basins: 3D/1D Amplification Ratios from Fast Multipole BEM Simulations

    CERN Document Server

    Fajardo, Kristel C Meza; Chaillat, Stéphanie; Lenti, Luca

    2016-01-01

    In this work, we study seismic wave amplification in alluvial basins having 3D standard geometries through the Fast Multipole Boundary Element Method in the frequency domain. We investigate how much 3D amplification differs from the 1D (horizontal layering) case. Considering incident fields of plane harmonic waves, we examine the relationships between the amplification level and the most relevant physical parameters of the problem (impedance contrast, 3D aspect ratio, vertical and oblique incidence of plane waves). The FMBEM results show that the most important parameters for wave amplification are the impedance contrast and the so-called equivalent shape ratio. Using these two parameters, we derive simple rules to compute the fundamental frequency for various 3D basin shapes and the corresponding 3D/1D amplification factor for 5% damping. Effects on amplification due to 3D basin asymmetry are also studied and incorporated in the derived rules.

  16. Relationship between Immunological Abnormalities in Rat Models of Diabetes Mellitus and the Amplification Circuits for Diabetes

    Science.gov (United States)

    Shimomura, Tomoko; Asao, Hironobu; Wakabayashi, Ichiro

    2017-01-01

    A better understanding of pathogenic mechanisms is required in order to treat diseases. However, the mechanisms of diabetes mellitus and diabetic complications are extremely complex. Immune reactions are involved in the pathogenesis of diabetes and its complications, while diabetes influences immune reactions. Furthermore, both diabetes and immune reactions are influenced by genetic and environmental factors. To address these issues, animal models are useful tools. So far, various animal models of diabetes have been developed in rats, which have advantages over mice models in terms of the larger volume of tissue samples and the variety of type 2 diabetes models. In this review, we introduce rat models of diabetes and summarize the immune reactions in diabetic rat models. Finally, we speculate on the relationship between immune reactions and diabetic episodes. For example, diabetes-prone Biobreeding rats, type 1 diabetes model rats, exhibit increased autoreactive cellular and inflammatory immune reactions, while Goto-Kakizaki rats, type 2 diabetes model rats, exhibit increased Th2 reactions and attenuation of phagocytic activity. Investigation of immunological abnormalities in various diabetic rat models is useful for elucidating complicated mechanisms in the pathophysiology of diabetes. Studying immunological alterations, such as predominance of Th1/17 or Th2 cells, humoral immunity, and innate immune reactions, may improve understanding the structure of amplification circuits for diabetes in future studies.

  17. 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.

  18. Generation of recombinant pestiviruses using a full genome amplification strategy

    DEFF Research Database (Denmark)

    Rasmussen, Thomas Bruun; Reimann, Ilona; Uttenthal, Åse

    pestiviruses. Methods Pestivirus genomes were amplified from either total RNA preparations using long RT-PCR or from infectious cDNA clones using long PCR. Viral RNA was extracted from cell cultures inoculated with pestivirus (e.g. BDV “Gifhorn” or BVDV “CP7”) using a combined Trizol/RNeasy protocol. Total RNA...... was reverse transcribed to cDNA at 50C for 90 minutes using SuperScript III reverse transcriptase (Invitrogen). Full-length PCR amplification was performed using primers specific for the extreme 5’- and 3’-ends of the viral genomes. A T7 promoter was incorporated in the 5’-primers for direct in vitro...... transcription of the amplicons. Long (RT)-PCR was performed using Accuprime High Fidelity or Elongase enzyme mix (Invitrogen), which consists of mixtures of Taq and proofreading Pyrococcus GB-D DNA polymerases. Reactions containing 2 l cDNA were amplified using 94C for 30 seconds followed by 35 cycles of 94°C...

  19. 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.)

  20. Emission-Line Galaxies from the Hubble Space Telescope Probing Evolution and Reionization Spectroscopically (PEARS) Grism Survey. II: The Complete Sample

    CERN Document Server

    Pirzkal, Nor; Ly, Chun; Malhotra, Sangeeta; Rhoads, James E; Grogin, Norman A; Dahlen, Tomas; Meurer, Gerhardt R; Walsh, Jeremy R; Hathi, Nimish P; Cohen, Seth H; Bellini, Andrea; Holwerda, Benne W; Straughn, Amber N; Mechtley, Matthew

    2012-01-01

    We present a full analysis of the Probing Evolution And Reionization Spectroscopically (PEARS) slitess grism spectroscopic data obtained with 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 1e9} M_sun decreases by an order of magnitude at z<0.5 relative to the number at 0.5

  1. Broadening and Amplification of an Infrared Femtosecond Pulse for Optical Parametric Chirped-Pulse Amplification

    Institute of Scientific and Technical Information of China (English)

    WANG He-Lin; YANG Ai-Jun; LENG Yu-Xin

    2011-01-01

    A high-average-power diode-pumped narrowband regenerative chirped pulse amplifier is developed using the thin-rod Nd:YAG laser architecture for optical parametric chirped-pulse amplification (OPCPA).The effect of the etalons on the amplified pulse in the regenerative cavity is studied experimentally and theoretically.By inserting glass etalons of thickness 1 mm and 5 mm into the regenerative cavity,the pre-stretching pulse from an (O)ffner stretcher is further broadened to above 200ps,which matches the amplification windows of the signal pulses in OPCPA and is suitable for use as a pump source in the OPCPA system.The bandwidth of the amplified pulse is 1.5 nm,and an output energy of 2mJ is achieved at a repetition rate of 10 Hz.Optical parametric chirped pulse amplification (OPCPA)[1-4] has attracted a great deal of attention as the most promising technique for generating ultrashort ultrahigh-peak-power laser pulses because of its very broad gain bandwidth,negligible thermal load on the nonlinear crystal,and extremely high singlepass gain as compared to amplifiers based on laser gain media.For efficient amplification and high fidelity of dispersion compensation in OPCPA,a femtosecond seed pulse is first stretched to several tens of picoseconds with a bulk grating stretcher or a fiber stretcher.%A high-average-power diode-pumped narrowband regenerative chirped pulse amplifier is developed using the thin-rod Nd:YAG laser architecture for optical parametric chirped-pulse amplification (OPCPA). The effect of the etalons on the amplified pulse in the regenerative cavity is studied experimentally and theoretically. By inserting glass etalons of thickness 1 mm and 5 mm into the regenerative cavity, the pre-stretching pulse from an (O)finer stretcher is further broadened to above 200 ps, which matches the amplification windows of the signal pulses in OPCPA and is suitable for use as a pump source in the OPCPA system. The bandwidth of the amplified pulse is 1.5 nm, and an

  2. Loop‑mediated Isothermal Amplification assay (LAMP) based detection of Pasteurella multocida in cases of haemorrhagic septicaemia and fowl cholera.

    Science.gov (United States)

    Bhimani, Mayurkumar; Bhanderi, Bharat; Roy, Ashish

    2015-01-01

    Twenty two isolates of Pasteurella multocida were obtained from different tissues of dead birds and animals (cattle, buffalo, sheep, and goat) suspected of fowl cholera and haemorrhagic septicaemia. The isolates were confirmed as P. multocida by various biochemical tests and PM PCR. An attempt was made to standardize Loop mediated isothermal amplification (LAMP) using newly designed primer sequences of KMT1 gene. Loop mediated isothermal amplification was conducted using 6 sets of primers at 65°C for 30 minutes and the result was confirmed by visual observation using SYBR green fluorescence dye as marker of positive reaction under UV transilluminator. On electrophoretic analysis of the products on 2% agarose gel, a ladder like pattern was observed, which suggested a positive amplification, whereas no amplification was observed in negative controls. Additionally, product of positive reaction yielded a green fluorescence following addition of SYBR green under UV transilluminator. It was observed that LAMP is a more sensitive test than polymerase chain reaction (PCR), as the former could detect DNA to lower limit of 22.8 pg/µl, while the latter could detect DNA to lower limit of 2.28 ng/ µl, thus LAMP could detect 100 times lesser concentration of DNA in comparison to PCR. Loop mediated isothermal amplification is a rather newer molecular technique, which can be used for rapid detection of infectious agent at field level and which does not require sophisticated instrument, i.e. thermal cycler. Furthermore, unlike the conventional PCR technique, LAMP requires lesser time to perform and result can be read visually.

  3. Label-free thioflavin T/G-quadruplex-based real-time strand displacement amplification for biosensing applications.

    Science.gov (United States)

    Du, Yi-Chen; Zhu, Li-Na; Kong, De-Ming

    2016-12-15

    To promote application of strand-displacement amplification (SDA) techniques in biosensing, a label-free, real-time monitoring strategy for isothermal nucleic acid amplification reactions was designed. G-quadruplex structures were introduced into SDA products using specific recognition of G-quadruplexes by the fluorogenic dye thioflavin T. Performance was good for real-time monitoring of traditional SDA by a linear-amplification mechanism and for exponential cross-triggered SDA amplification. The strategy worked on a commercial real-time PCR instrument, making it suitable for biosensing platforms. As examples, two highly sensitive and specific biosensors were designed for analysis of the activity of uracil-DNA glycosylase (UDG) and the restriction endonuclease EcoRI. Detection limits were 6×10(-5)U/mL for UDG and 0.016U/mL for EcoRI. Detection of corresponding targets in complex matrices such as cell lysates or human serum was also demonstrated. Compared to traditional end-point detection methods, real-time SDA-based approaches have the advantages of simple, fast operation; high sensitivity; low risk of carryover contamination; and very high throughput. The introduction of real-time monitoring strategies may promote application of SDA reactions in biosensor design.

  4. In vitro morphogenetic response of apple (Malus domestica Borkh. and pear (Pyrus communis L. to the elevated levels of copper and myo-inositol

    Directory of Open Access Journals (Sweden)

    Rafail S. Toma

    2012-10-01

    Full Text Available The elevated levels of copper and myo-inositol in the MS medium were demonstrated to enhance culture growth and morphogenetic response of apple and pear explants. The results revealed that the highest number of branches per explant (2.80 for apple was obtained from the levels of 0.0+ 100 and 0.050+400 mg/l of both copper and myo-inositol, respectively (C1M2 and C4M4, while for pear 3.40 branches per explant were achieved from the same treatment. The mean length of branches was significantly lower in the case of the control treatment (the absence of copper and inositol. The highest number of leaves per explant (29.73 and 29.80 for both apple and pear, respectively, was recorded for treatment C4M4 (0.050+ 400 mg/l of both copper and myo-inositol, respectively. At the rooting stage, the elevated levels of copper and myo-inositol were very effective in stimulating root formation in both apple and pear shoots. The highest number of roots in apple (2.00 roots/ explant was achieved while using 0.100+ 800 (C5M5 of both copper and myo-inositol, whereas the highest number of roots for pear (3.17 roots/ explant was recorded for C6M6 (0.200+ 1600. The highest mean length of roots for apple reached 1.23 cm in treatment C3M3 and 1.10 cm for pear in treatment C6M6. These data suggest that the higher levels of copper and myo-inositol enabled shoot and root formation in the explants, and it might be necessary to use higher levels of these two medium components in order to enhance morphogenetic potential of explants.

  5. Characterization of CIPK family in Asian pear (Pyrus bretschneideri Rehd and co-expression analysis related to salt and osmotic stress responses

    Directory of Open Access Journals (Sweden)

    Jun Tang

    2016-09-01

    Full Text Available Asian pear (Pyrus bretschneideri is one of the most important fruit crops in the world, and its growth and productivity are frequently affected by abiotic stresses. Calcineurin B-like interacting protein kinases (CIPKs as caladium-sensor protein kinases interact with Ca2+-binding CBLs to extensively mediate abiotic stress responses in plants. Although the pear genome sequence has been released, little information is available about the CIPK genes in pear, especially in response to salt and osmotic stresses. In this study, we systematically identified 28 CIPK family members from the sequenced pear genome and analyzed their organization, phylogeny, gene structure, protein motif, and synteny duplication divergences. Most duplicated PbCIPKs underwent purifying selection, and their evolutionary divergences accompanied with the pear whole genome duplication. We also investigated stress -responsive expression patterns and co-expression networks of CIPK family under salt and osmotic stresses, and the distribution of stress-related cis-regulatory elements in promoter regions. Our results suggest that most PbCIPKs could play important roles in the abiotic stress responses. Some PbCIPKs, such as PbCIPK22, -19, -18, -15, -8, and -6 can serve as core regulators in response to salt and osmotic stresses based on co-expression networks of PbCIPKs. Some sets of genes that were involved in response to salt did not overlap with those in response to osmotic responses, suggesting the sub-functionalization of CIPK genes in stress responses. This study revealed some candidate genes that play roles in early responses to salt and osmotic stress for further characterization of abiotic stress responses medicated by CIPKs in pear.

  6. 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.

  7. Multiplex isothermal solid-phase recombinase polymerase amplification for the specific and fast DNA-based detection of three bacterial pathogens.

    Science.gov (United States)

    Kersting, Sebastian; Rausch, Valentina; Bier, Frank F; von Nickisch-Rosenegk, Markus

    2014-01-01

    We report on the development of an on-chip RPA (recombinase polymerase amplification) with simultaneous multiplex isothermal amplification and detection on a solid surface. The isothermal RPA was applied to amplify specific target sequences from the pathogens Neisseria gonorrhoeae, Salmonella enterica and methicillin-resistant Staphylococcus aureus (MRSA) using genomic DNA. Additionally, a positive plasmid control was established as an internal control. The four targets were amplified simultaneously in a quadruplex reaction. The amplicon is labeled during on-chip RPA by reverse oligonucleotide primers coupled to a fluorophore. Both amplification and spatially resolved signal generation take place on immobilized forward primers bount to expoxy-silanized glass surfaces in a pump-driven hybridization chamber. The combination of microarray technology and sensitive isothermal nucleic acid amplification at 38 °C allows for a multiparameter analysis on a rather small area. The on-chip RPA was characterized in terms of reaction time, sensitivity and inhibitory conditions. A successful enzymatic reaction is completed in isothermal nucleic acid amplification with RPA and spatially-resolved signal generation on specific immobilized oligonucleotides.

  8. Control and amplification of cortical neurodynamics

    Science.gov (United States)

    Liljenstroem, Hans; Aronsson, P.

    1999-03-01

    We investigate different mechanisms for the control and amplification of cortical neurodynamics, using a neural network model of a three layered cortical structure. We show that different dynamical states can be obtained by changing a control parameter of the input-output relation, or by changing the noise level. Point attractor, limit cycle, and strange attractor dynamics occur at different values of the control parameter. For certain, optimal noise levels, system performance is maximized, analogous to stochastic resonance phenomena. Noise can also be used to induce different dynamical states. A few noisy network units distributed in a network layer can result in global synchronous oscillations, or waves of activity moving across the network. We further demonstrate that fast synchronization of network activity can be obtained by implementing electromagnetic interactions between network units.

  9. Amplification sans bruit d'images optiques

    Science.gov (United States)

    Gigan, S.; Delaubert, V.; Lopez, L.; Treps, N.; Maitre, A.; Fabre, C.

    2004-11-01

    Nous utilisons un Oscillateur Paramétrique Optique (OPO) pompé sous le seuil dans le but d'amplifier une image multimode transverse sans dégradation du rapport signal à bruit. Le dispositif expérimental met en œuvre un OPO de type II triplement résonant et semi-confocal pour le faisceau amplifié. L'existence d'effets quantiques lors de l'amplification multimode dans un tel dispositif a été montrée expérimentalement. Plus généralement, ceci nous a amené à étudier les propriétés quantiques transverses des faisceaux lumineux amplifiés. Une telle étude peut trouver des applications non seulement en imagerie, mais également dans le traitement quantique de l'information.

  10. Dispersion compensation in chirped pulse amplification systems

    Science.gov (United States)

    Bayramian, Andrew James; Molander, William A.

    2014-07-15

    A chirped pulse amplification system includes a laser source providing an input laser pulse along an optical path. The input laser pulse is characterized by a first temporal duration. The system also includes a multi-pass pulse stretcher disposed along the optical path. The multi-pass pulse stretcher includes a first set of mirrors operable to receive input light in a first plane and output light in a second plane parallel to the first plane and a first diffraction grating. The pulse stretcher also includes a second set of mirrors operable to receive light diffracted from the first diffraction grating and a second diffraction grating. The pulse stretcher further includes a reflective element operable to reflect light diffracted from the second diffraction grating. The system further includes an amplifier, a pulse compressor, and a passive dispersion compensator disposed along the optical path.

  11. Magnetic field amplification in turbulent astrophysical plasmas

    CERN Document Server

    Federrath, Christoph

    2016-01-01

    Magnetic fields play an important role in astrophysical accretion discs, and in the interstellar and intergalactic medium. They drive jets, suppress fragmentation in star-forming clouds and can have a significant impact on the accretion rate of stars. However, the exact amplification mechanisms of cosmic magnetic fields remain relatively poorly understood. Here I start by reviewing recent advances in the numerical and theoretical modelling of the 'turbulent dynamo', which may explain the origin of galactic and inter-galactic magnetic fields. While dynamo action was previously investigated in great detail for incompressible plasmas, I here place particular emphasis on highly compressible astrophysical plasmas, which are characterised by strong density fluctuations and shocks, such as the interstellar medium. I find that dynamo action works not only in subsonic plasmas, but also in highly supersonic, compressible plasmas, as well as for low and high magnetic Prandtl numbers. I further present new numerical simu...

  12. Anisotropic metamaterials with simultaneous attenuation and amplification

    CERN Document Server

    Mackay, Tom G

    2015-01-01

    Anisotropic metamaterials that are neither wholly dissipative nor wholly active at a specific frequency are permitted by classical electromagnetic theory. Well-established formalisms for the homogenization of particulate composite materials indicate that such a metamaterial may be conceptualized quite simply as a random mixture of electrically small spheroidal particles of at least two different isotropic dielectric materials, one of which must be dissipative but the other active. The realization of this metametarial is influenced by the volume fraction, spatial distribution, particle shape and size, and the relative permittivities of the component materials. Metamaterials displaying both dissipation and amplification at the same frequency with more complicated linear as well as nonlinear constitutive properties are possible.

  13. Integrated Amplification Microarrays for Infectious Disease Diagnostics

    Directory of Open Access Journals (Sweden)

    Darrell P. Chandler

    2012-11-01

    Full Text Available This overview describes microarray-based tests that combine solution-phase amplification chemistry and microarray hybridization within a single microfluidic chamber. The integrated biochemical approach improves microarray workflow for diagnostic applications by reducing the number of steps and minimizing the potential for sample or amplicon cross-contamination. Examples described herein illustrate a basic, integrated approach for DNA and RNA genomes, and a simple consumable architecture for incorporating wash steps while retaining an entirely closed system. It is anticipated that integrated microarray biochemistry will provide an opportunity to significantly reduce the complexity and cost of microarray consumables, equipment, and workflow, which in turn will enable a broader spectrum of users to exploit the intrinsic multiplexing power of microarrays for infectious disease diagnostics.

  14. Magnetic Field Amplification and Blazar Flares

    CERN Document Server

    Chen, Xuhui; Fossati, Giovanni; Pohl, Martin

    2013-01-01

    Recent multiwavelength observations of PKS 0208-512 by SMARTS, Fermi, and Swift revealed that gamma-ray and optical light curves of this flat spectrum radio quasars are highly correlated, but with an exception of one large optical flare having no corresponding gamma-ray activity or even detection. On the other hand, recent advances in SNRs observations and plasma simulations both reveal that magnetic field downstream of astrophysical shocks can be largely amplified beyond simple shock compression. These amplifications, along with their associated particle acceleration, might contribute to blazar flares, including the peculiar flare of PKS 0208-512. Using our time dependent multizone blazar emission code, we evaluate several scenarios that may represent such phenomena. This code combines Monte Carlo method that tracks the radiative processes including inverse Compton scattering, and Fokker-Planck equation that follows the cooling and acceleration of particles. It is a comprehensive time dependent code that ful...

  15. Mutation of the Erwinia amylovora argD gene causes arginine auxotrophy, nonpathogenicity in apples, and reduced virulence in pears.

    Science.gov (United States)

    Ramos, Laura S; Lehman, Brian L; Peter, Kari A; McNellis, Timothy W

    2014-11-01

    Fire blight is caused by Erwinia amylovora and is the most destructive bacterial disease of apples and pears worldwide. In this study, we found that E. amylovora argD(1000)::Tn5, an argD Tn5 transposon mutant that has the Tn5 transposon inserted after nucleotide 999 in the argD gene-coding region, was an arginine auxotroph that did not cause fire blight in apple and had reduced virulence in immature pear fruits. The E. amylovora argD gene encodes a predicted N-acetylornithine aminotransferase enzyme, which is involved in the production of the amino acid arginine. A plasmid-borne copy of the wild-type argD gene complemented both the nonpathogenic and the arginine auxotrophic phenotypes of the argD(1000)::Tn5 mutant. However, even when mixed with virulent E. amylovora cells and inoculated onto immature apple fruit, the argD(1000)::Tn5 mutant still failed to grow, while the virulent strain grew and caused disease. Furthermore, the pCR2.1-argD complementation plasmid was stably maintained in the argD(1000)::Tn5 mutant growing in host tissues without any antibiotic selection. Therefore, the pCR2.1-argD complementation plasmid could be useful for the expression of genes, markers, and reporters in E. amylovora growing in planta, without concern about losing the plasmid over time. The ArgD protein cannot be considered an E. amylovora virulence factor because the argD(1000)::Tn5 mutant was auxotrophic and had a primary metabolism defect. Nevertheless, these results are informative about the parasitic nature of the fire blight disease interaction, since they indicate that E. amylovora cannot obtain sufficient arginine from apple and pear fruit tissues or from apple vegetative tissues, either at the beginning of the infection process or after the infection has progressed to an advanced state.

  16. Over-expression of the apple spermidine synthase gene in pear confers multiple abiotic stress tolerance by altering polyamine titers.

    Science.gov (United States)

    Wen, Xiao-Peng; Pang, Xiao-Ming; Matsuda, Narumi; Kita, Masayuki; Inoue, Hiromichi; Hao, Yu-Jin; Honda, Chikako; Moriguchi, Takaya

    2008-04-01

    An apple spermidine synthase (SPDS) gene (MdSPDS1) was verified to encode a functional protein by the complementation of the spe3 yeast mutant, which lacks the SPDS gene. To justify our hypothesis that apple SPDS is involved in abiotic stress responses and to obtain transgenic fruit trees tolerant to abiotic stresses as well, MdSPDS1-over-expressing transgenic European pear (Pyrus communis L. 'Ballad') plants were created by Agrobacterium-mediated transformation. A total of 21 transgenic lines showing various spermidine (Spd) titers and MdSPDS1 expression levels were obtained. Selected lines were exposed to salt (150 mM NaCl), osmosis (300 mM mannitol), and heavy metal (500 microM CuSO4) stresses for evaluating their stress tolerances. Transgenic line no. 32, which was revealed to have the highest Spd accumulation and expression level of MdSPDS1, showed the strongest tolerance to these stresses. When growth increments, electrolyte leakage (EL), and values of thiobarbituric acid reactive substances (TBARS) were monitored, line no. 32 showed the lowest growth inhibition and the least increase in EL or TBARS under stress conditions. Spd titers in wild-type and transgenic lines showed diverse changes upon stresses, and these changes were not consistent with the changes in MdSPDS1 expressions. Moreover, there were no differences in the sodium concentration in the shoots between the wild type and line no. 32, whereas the copper concentration was higher in the wild type than in line no. 32. Although the mechanism(s) underlying the involvement of polyamines in stress responses is not known, these results suggest that the over-expression of the SPDS gene substantially increased the tolerance to multiple stresses by altering the polyamine titers in pear. Thus, MdSPDS1-over-expressing transgenic pear plants could be used to improve desert land and/or to repair polluted environments.

  17. Oxidative stress associated with rootstock-scion interactions in pear/quince combinations during early stages of graft development.

    Science.gov (United States)

    Irisarri, Patricia; Binczycki, Piotr; Errea, Pilar; Martens, Helle Juel; Pina, Ana

    2015-03-15

    Exposing a plant to stress situations, such as grafting, generally triggers antioxidant defense systems. In fruit tree grafting, quince (Cydonia oblonga) is widely used as a rootstock for pear (Pyrus communis L.), but several economically important pear cultivars are incompatible with available quince rootstocks. In this study, grafts were established using an in vitro callus graft system mimicking the events taking place in fruit trees. In vitro grown callus from pear [P. communis L. cv. 'Conference' (Co) and cv. 'William' (Wi)] and quince (C. oblonga Mill. clone 'BA29') was used to establish the compatible homografts 'Co/Co', 'Wi/Wi' and 'BA29/BA29', the compatible heterograft 'Co/BA29' and the incompatible heterograft 'Wi/BA29'. The main objective was to determine whether specific isoforms of genes involved in oxidative stress [superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT)] are differentially expressed at the graft interface from compatible and incompatible unions throughout 3 weeks after grafting. Reactive oxygen species (ROS) levels and programmed cell death were also evaluated in the course of graft development. Genes differentially expressed between compatible and incompatible heterografts were identified. Transcript levels of six antioxidant genes (SOD1, SOD3, APX3, APX6, CAT1 and CAT3) were down-regulated 10 days after grafting (DAG) in the incompatible heterograft in comparison to the compatible one. Likewise, SOD enzymatic activities were significantly higher at 1 and 10 days after wounding in the compatible cultivar 'Co' than in the incompatible one 'Wi'. These findings, together with live cell imaging of ROS-specific probes, ultrastructural mitochondrial changes and DNA fragmentation related to apoptotic processes, give indications that within incompatible rootstock/scion interfaces, either the level of ROS is increased or there is a less efficient detoxification system.

  18. Occurrence of indole-3-acetic Acid-producing bacteria on pear trees and their association with fruit russet.

    Science.gov (United States)

    Lindow, S E; Desurmont, C; Elkins, R; McGourty, G; Clark, E; Brandl, M T

    1998-11-01

    ABSTRACT A relatively high percentage of epiphytic bacteria on pear leaf and fruit surfaces had the ability to produce indole-3-acetic acid (IAA) in culture media supplemented with tryptophan. While over 50% of the strains produced at least small amounts of IAA in culture, about 25% of the strains exhibited high IAA production as evidenced by both colorimetric and high-performance liquid chromatography analysis of culture supernatants. A majority of the strains that produced high amounts of IAA were identified as Erwinia herbicola (Pantoea agglomerans), while some strains of Pseudomonas syringae, Pseudomonas viridiflava, Pseudomonas fluorescens, Pseudomonas putida, and Rahnella aquaticus that produced high amounts of IAA also were found on pear. Fruit russeting was significantly increased in 39 out of 46 trials over an 8-year period in which IAA-producing bacteria were applied to trees compared with control trees. A linear relationship was observed between fruit russet severity and the logarithm of the population size of different IAA-producing bacteria on trees in the 30 days after inoculation, when normalized for the amount of IAA produced by each strain in culture. On average, the severity of fruit russet was only about 77% that on control trees when trees were treated at the time of bloom with Pseudomonas fluorescens strain A506, which does not produce IAA. Both total bacterial populations on pear in the 30-day period following full bloom and fruit russet severity varied greatly from year to year and in different commercial orchards over a 10-year period. There was a strong linear correlation between the logarithm of total bacterial population sizes and fruit russet severity.

  19. Short-Pulse Amplification by Strongly-Coupled Brillouin Scattering

    CERN Document Server

    Edwards, Matthew R; Mikhailova, Julia M; Fisch, Nathaniel J

    2016-01-01

    We examine the feasibility of strongly-coupled stimulated Brillouin scattering as a mechanism for the plasma-based amplification of sub-picosecond pulses. In particular, we use fluid theory and particle-in-cell simulations to compare the relative advantages of Raman and Brillouin amplification over a broad range of achievable parameters.

  20. A Theoretical Evaluation of Optical Parametric Amplification in BBO Crystal

    Institute of Scientific and Technical Information of China (English)

    邵敏; 薛绍林; 林尊琪

    2005-01-01

    The noncollinear optical parametric amplification in BBO crystal is theoretically investigated. The phase matching angle, gain bandwidth, optimal noncollinear angle and conversion efficiency for both type-Ⅰ and type-Ⅱ BBO are simulated. The numerical simulation results are important to the practical optical parametric amplification experiments with BBO crystal.

  1. Targeting HER2 amplifications in gastric cancer

    Directory of Open Access Journals (Sweden)

    Ung L

    2014-01-01

    Full Text Available Lawson Ung, Terence C Chua, Neil D Merrett Department of Surgery, South Western Sydney Upper GI Surgical Unit, Bankstown Hospital, University of Western Sydney, Sydney, NSW, Australia Abstract: While multimodality treatments, including neoadjuvant and adjuvant chemotherapy or chemoradiation, have become the global standard of care in patients with locally advanced and metastatic gastric cancers (GCs, long-term outcomes for patients remain poor. This reflects the aggressive tumor biology of GCs and occult nature of the disease, often presenting in its advanced stages, as well as the challenges of developing effective targeted therapy to treat this disease. The Trastuzumab for Gastric Cancer trial demonstrates that the addition of human epidermal growth factor 2 (HER2 monoclonal antibody trastuzumab to standard chemotherapy regimen consisting of 5-fluorouracil (5-FU or capecitabine with cisplatin results in significant improvement in overall and progression-free survival. Although questions remain regarding the best methods by which to determine HER2 mutation positivity and amplification, through immunohistochemistry or in situ hybridization, and whether trastuzumab is effective for locally advanced, nonmetastatic GC in an adjuvant setting, the trial has led to a surge of clinical trials investigating the potential role of other HER2- and non-HER2-targeted therapies to improve patient outcomes. This review will discuss our current understanding of GC pathogenesis, current available treatments, and the potential impact that targeting HER2 amplifications may have in our efforts to individualize and optimize cancer care in GC individuals. Keywords: Personalized cancer therapy, surgical oncology, gastrectomy, adjuvant treatment, targeted therapies

  2. Mutualism breakdown by amplification of Wolbachia genes.

    Science.gov (United States)

    Chrostek, Ewa; Teixeira, Luis

    2015-02-01

    Most insect species are associated with vertically transmitted endosymbionts. Because of the mode of transmission, the fitness of these symbionts is dependent on the fitness of the hosts. Therefore, these endosymbionts need to control their proliferation in order to minimize their cost for the host. The genetic bases and mechanisms of this regulation remain largely undetermined. The maternally inherited bacteria of the genus Wolbachia are the most common endosymbionts of insects, providing some of them with fitness benefits. In Drosophila melanogaster, Wolbachia wMelPop is a unique virulent variant that proliferates massively in the hosts and shortens their lifespan. The genetic bases of wMelPop virulence are unknown, and their identification would allow a better understanding of how Wolbachia levels are regulated. Here we show that amplification of a region containing eight Wolbachia genes, called Octomom, is responsible for wMelPop virulence. Using Drosophila lines selected for carrying Wolbachia with different Octomom copy numbers, we demonstrate that the number of Octomom copies determines Wolbachia titers and the strength of the lethal phenotype. Octomom amplification is unstable, and reversion of copy number to one reverts all the phenotypes. Our results provide a link between genotype and phenotype in Wolbachia and identify a genomic region regulating Wolbachia proliferation. We also prove that these bacteria can evolve rapidly. Rapid evolution by changes in gene copy number may be common in endosymbionts with a high number of mobile elements and other repeated regions. Understanding wMelPop pathogenicity and variability also allows researchers to better control and predict the outcome of releasing mosquitoes transinfected with this variant to block human vector-borne diseases. Our results show that transition from a mutualist to a pathogen may occur because of a single genomic change in the endosymbiont. This implies that there must be constant selection on

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

    OpenAIRE

    Wenneker, M.; Pham, K.T.K.; Lemmers, M.E.C.; De Boer, MR; Leeuwen, van, M.; 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 agar, and at low temperatures development of a fast-growing whitish mycelium was observed. Growth of this fungus was observed between 1 and 20 °C with an optimum at 15 °C, while incubation of mycelium a...

  4. 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 orchard environment.

  5. Promotion of flowering by Apple latent spherical virus vector and virus elimination at high temperature allow accelerated breeding of apple and pear

    Directory of Open Access Journals (Sweden)

    Noriko eYamagishi

    2016-02-01

    Full Text Available Plant viral vectors are superior tools for genetic manipulation, allowing rapid induction or suppression of expression of a target gene in plants. This is a particularly effective technology for use in breeding fruit trees, which are difficult to manipulate using recombinant DNA technologies. We reported previously that if apple seed embryos (cotyledons are infected with an Apple latent spherical virus (ALSV vector (ALSV-AtFT/MdTFL1 concurrently expressing the Arabidopsis thaliana florigen (AtFT gene and suppressing the expression of the apple MdTFL1-1 gene, the period prior to initial flowering (generally lasts 5–12 years will be reduced to about two months. In this study, we examined whether or not ALSV vector technology can be used to promote flowering in pear, which undergoes a very long juvenile period (germination to flowering similar to that of apple. The MdTFL1 sequence in ALSV-AtFT/MdTFL1 was replaced with a portion of the pear PcTFL1-1 gene. The resulting virus (ALSV-AtFT/PcTFL1 and ALSV-AtFT/MdTFL1 were used individually for inoculation to pear cotyledons immediately after germination in two inoculation groups. Those inoculated with ALSV-AtFT/MdTFL1 and ALSV-AtFT/PcTFL1 then initiated flower bud formation starting one to three months after inoculation, and subsequently exhibited continuous flowering and fruition by pollination. Conversely, Japanese pear exhibited extremely low systemic infection rates when inoculated with ALSV-AtFT/MdTFL1, and failed to exhibit any induction of flowering. We also developed a simple method for eliminating ALSV vectors from infected plants. An evaluation of the method for eliminating the ALSV vectors from infected apple and pear seedlings revealed that a four-week high-temperature (37˚C incubation of ALSV-infected apples and pears disabled the movement of ALSV to new growing tissues. This demonstrates that only high-temperature treatment can easily eliminate ALSV from infected fruit trees. A method

  6. 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.

  7. [Nondestructive measurement of SSC in western pear using genetic algorithms and FT-NIR spectroscopy].

    Science.gov (United States)

    Wang, Jia-Hua; Pan, Lu; Sun, Qian; Li, Peng-Fei; Han, Dong-Hai

    2009-03-01

    An improved genetic algorithm was used to implement an automated wavelength selection procedure for use in building multivariate calibration models based on partial least squares regression (PLS). The region selecting by genetic algorithms (R-SGA) was applied in building calibration model of soluble solid content (SSC) of Western pear, and the numbers of latent variables used to build calibration model were further reduced. The Fourier transform near infrared reflectance (FT-NIR) spectra were processed by GA after MSC or SNV, and four PLS calibration models were built by using the optimal combinations of these sub-regions. Meanwhile, the full region selecting PLS (Fr-PLS) models were developed. The R-SGA models variables were 434, 496, 310 and 496, for Early Red Comice, Wujiuxiang, Cascade and Kang Buddha, respectively. Despite the complexity of the spectral data, the R-SGA procedure was found to perform well (RMSEP = 0.428, 0.567 for Early Red Comice and Kang Buddha, respectively), leading to calibration models that significantly outperform those based on full-spectrum analyses (RM-SEP = 0.518, 0.633). The prediction precision of GA-PLS models was similar to that of Fr-PLS for Wujiuxiang and Cascade, with RMSEP of 0.696/0.694 and 0.425/0.421 respectively. This work proved that the R-SGA could find optimal values for several disparate variables associated with the calibration model and that the PLS procedure could be integrated into the objective function driving the optimization.

  8. Autonomous Biological Control of Dactylopius opuntiae (Hemiptera: Dactyliiopidae) in a Prickly Pear Plantation With Ecological Management.

    Science.gov (United States)

    Cruz-Rodríguez, J A; González-Machorro, E; Villegas González, A A; Rodríguez Ramírez, M L; Mejía Lara, F

    2016-04-07

    It is broadly known that the conservation of biological diversity in agricultural ecosystems contributes to pest control. This process was studied in a prickly pear plantation (Opuntia megacanthaandOpuntia ficus-indica) located in central Mexico. No insecticides have been used on this plantation since 2000, and local farmers believe that the presence of different species of insects limits the growth of the wild cochineal (Dactylopius opuntiaeCockerell), which is one of the main pests in this crop. From August 2012 to November 2013, we estimated the number of cochineal per stem in the plantation and determined its spatial distribution pattern. In order to identify signs of population regulation, we obtained histograms of the frequency distribution of the size of the clusters and determined if distribution is adjusted to a power function (power law). We identified the cochineal predators and determined the correlation in their abundances. The greater abundance of cochineal occurred between summer and autumn while the minimum value was recorded in spring. The frequency distribution of the cochineal clusters had a high level of adjustment to a power function, suggesting the presence of population regulation processes. Six species that prey on cochineal were identified.Laetilia coccidivoraandHyperaspis trifurcatawere the most active and their abundance was significantly correlated with the abundance of cochineal. We found that the probability of extinction of these insects in a cladode increases with its density, since the density and predator activity also increased. It is likely that, under these conditions, the cochineal have established an autonomous control.

  9. Parametric Analog Signal Amplification Applied to Nanoscale CMOS Technologies

    CERN Document Server

    Oliveira, João P

    2012-01-01

    This book is dedicated to the analysis of parametric amplification with special emphasis on the MOS discrete-time implementation. This implementation is demonstrated by the presentation of several circuits where the MOS parametric amplifier cell is used: small gain amplifier, comparator with embedded pre-amplification, discrete-time mixer/IIR-Filter, and analog-to-digital converter (ADC).  Experimental results are shown to validate the overall design technique. Provides the complete theoretical analysis, supported by electrical simulations, of the parametric amplification technique in both continuous time and discrete time domains; Describes the design flow of an ADC fully based on discrete-time parametric amplification in CMOS technology; Presents a high speed time-interleaved pipeline ADC, based on parametric MOS amplification techniques described, complementing theory discussed with experimental results.

  10. Development of loop-mediated isothermal amplification for rapid detection of Entamoeba histolytica

    Institute of Scientific and Technical Information of China (English)

    Windell L Rivera; Vanissa A Ong

    2013-01-01

    Objective: To develop a loop-mediated isothermal amplification (LAMP) assay for the detection of Entamoeba histolytica (E. histolytica), the causative agent of amebiasis. Methods: The LAMP primer set was designed from E. histolytica hemolysin gene HLY6. Genomic DNA of E. histolytica trophozoites strain HK9 was used to optimize the LAMP mixture and conditions. Amplification of DNA in the LAMP mixture was monitored through visual inspection for turbidity of the LAMP mix as well as addition of fluorescent dye. Results: Positive LAMP reactions turned turbid while negative ones remained clear. Upon addition of a fluorescent dye, all positive reactions turned green while the negative control remained orange under ambient light. After elecrophoresis in 1.5%agarose gels, a ladder of multiple bands of different sizes can be observed in positive samples while no bands were detected in the negative control. The sensitivity of the assay was found to be 5 parasites per reaction which corresponds to approximately 15.8 ng/μL DNA. The specificity of the assay was verified by the absence of amplified products when DNA from other gastrointestinal parasites such as the morphologically similar but non-pathogenic species, Entamoeba dispar, and other diarrhea-causing organisms such as Blastocystis hominis and Escherichia coli were used. Conclusions: The LAMP assay we have developed enables the detection of E. histolytica with rapidity and ease, therefore rendering it is suitable for laboratory and field diagnosis of amebiasis.

  11. 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.

  12. RNA amplification of bromodeoxyuridine labeled newborn neurons in the monkey hippocampus.

    Science.gov (United States)

    Counts, Scott E; Chen, Er-Yun; Ginsberg, Stephen D; Kordower, Jeffrey H; Mufson, Elliott J

    2005-06-15

    Neurogenesis has been demonstrated in the adult mammalian hippocampus by the immunohistochemical identification of cells co-labeled with the neuronal marker NeuN and bromodeoxyuridine (BrdU), a marker for DNA synthesis. Whether these newly born neurons exhibit a genetic signature similar to that of existing hippocampal cells remains unknown. Recent advances in single cell RNA amplification techniques provide a unique method for profiling the mRNA complement of cells developed during adult neurogenesis. Standard protocols for identifying BrdU-positive cells requires an acid denaturation step that may preclude the amplification of cellular RNA for expression analysis. We first tested whether the BrdU reaction product was visible in monkey hippocampal tissue following treatment with dilutions of HCl (2-0.2 M) or citric acid (1.0-0.1 M). BrdU-labeled cells were visible only in tissue sections treated with 2 M HCl. RNA amplification was not compromised in cells dual-labeled for BrdU and NeuN using the 2 M HCl acid denaturation step. These cells express mRNAs encoding a wide variety of functional protein subclasses including glutamate receptors. The present study demonstrates for the first time that BrdU immunohistochemisty is compatable with gene array technology in the primate hippocampus to evaluate subclasses of genes in newborn neurons.

  13. 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.

  14. Chemiluminescence imaging for microRNA detection based on cascade exponential isothermal amplification machinery.

    Science.gov (United States)

    Xu, Yongjie; Li, Dandan; Cheng, Wei; Hu, Rong; Sang, Ye; Yin, Yibing; Ding, Shijia; Ju, Huangxian

    2016-09-14

    A novel G-quadruplex DNAzyme-driven chemiluminescence (CL) imaging method was developed for ultrasensitive and specific detection of miRNA based on the cascade exponential isothermal amplification reaction (EXPAR) machinery. A structurally tailored hairpin probe switch was designed to selectively recognise miRNA and form hybridisation products to trigger polymerase and nicking enzyme machinery, resulting in the generation of product I, which was complementary to a region of the functional linear template. Then, the response of the functional linear template to the generated product I further activated the exponential isothermal amplification machinery, leading to synthesis of numerous horseradish peroxidase mimicking DNAzyme units for CL signal transduction. The amplification paradigm generated a linear response from 10 fM to 100 pM, with a low detection limit of 2.91 fM, and enabled discrimination of target miRNA from a single-base mismatched target. The developed biosensing platform demonstrated the advantages of isothermal, homogeneous, visual detection for miRNA assays, offering a promising tool for clinical diagnosis.

  15. Real-time quantitative nicking endonuclease-mediated isothermal amplification with small molecular beacons.

    Science.gov (United States)

    Xu, Wentao; Wang, Chenguang; Zhu, Pengyu; Guo, Tianxiao; Xu, Yuancong; Huang, Kunlun; Luo, Yunbo

    2016-04-21

    Techniques of isothermal amplification have recently made great strides, and have generated significant interest in the field of point-of-care detection. Nicking endonuclease-mediated isothermal amplification (NEMA) is an example of simple isothermal technology. In this paper, a real-time quantitative nicking endonuclease-mediated isothermal amplification with small molecular beacons (SMB-NEMA) of improved specificity and sensitivity is described. First, we optimized the prohibition of de novo synthesis by choosing Nt·BstNBI endonuclease. Second, the whole genome was successfully amplified with Nt·BstNBI (6 U), betaine (1 M) and trehalose (60 mM) for the first time. Third, we achieved 10 pg sensitivity for the first time after adding a small molecular beacon that spontaneously undergoes a conformational change when hybridizing to target, and the practical test validated the assay's application. The small molecular beacon has a similar melting temperature to the reaction temperature, but is approximately 10 bp shorter than the length of a traditional molecular beacon. A new threshold regulation was also established for isothermal conditions. Finally, we established a thermodynamic model for designing small molecular beacons. This multistate model is more correct than the traditional algorithm. This theoretical and practical basis will help us to monitor SMB-NEMA in a quantitative way. In summary, our SMB-NEMA method allows the simple, specific and sensitive assessment of isothermal DNA quantification.

  16. Whole genome amplification and de novo assembly of single bacterial cells.

    Directory of Open Access Journals (Sweden)

    Sébastien Rodrigue

    Full Text Available BACKGROUND: Single-cell genome sequencing has the potential to allow the in-depth exploration of the vast genetic diversity found in uncultured microbes. We used the marine cyanobacterium Prochlorococcus as a model system for addressing important challenges facing high-throughput whole genome amplification (WGA and complete genome sequencing of individual cells. METHODOLOGY/PRINCIPAL FINDINGS: We describe a pipeline that enables single-cell WGA on hundreds of cells at a time while virtually eliminating non-target DNA from the reactions. We further developed a post-amplification normalization procedure that mitigates extreme variations in sequencing coverage associated with multiple displacement amplification (MDA, and demonstrated that the procedure increased sequencing efficiency and facilitated genome assembly. We report genome recovery as high as 99.6% with reference-guided assembly, and 95% with de novo assembly starting from a single cell. We also analyzed the impact of chimera formation during MDA on de novo assembly, and discuss strategies to minimize the presence of incorrectly joined regions in contigs. CONCLUSIONS/SIGNIFICANCE: The methods describe in this paper will be useful for sequencing genomes of individual cells from a variety of samples.

  17. Recombinase polymerase amplification as a promising tool in hepatitis C virus diagnosis

    Institute of Scientific and Technical Information of China (English)

    Hosam; Zaghloul; Mahmoud; El-shahat

    2014-01-01

    Hepatitis C virus(HCV)infection represents a significant health problem and represents a heavy load on some countries like Egypt in which about 20%of the total population are infected.Initial infection is usually asymptomatic and result in chronic hepatitis that give rise to complications including cirrhosis and hepatocellular carcinoma.The management of HCV infection should not only be focus on therapy,but also to screen carrier individuals in order to prevent transmission.In the present,molecular detection and quantification of HCV genome by real time polymerase chain reaction(PCR)represent the gold standard in HCV diagnosis and plays a crucial role in the management of therapeutic regimens.However,real time PCR is a complicated approach and of limited distribution.On the other hand,isothermal DNA amplification techniques have been developed and offer molecular diagnosis of infectious dieses at point-of-care.In this review we discuss recombinase polymerase amplification technique and illustrate its diagnostic value over both PCR and other isothermal amplification techniques.

  18. Establishment and Optimization of SRAP Amplification System in Lonicara caerulea L.

    Institute of Scientific and Technical Information of China (English)

    SUN Feng; HUO Junwei; QIN Dong

    2011-01-01

    A single factor design was applied to optimize five factors influencing SRAP system, including Taq DNA polymerase, template DNA concentration, dNTPs, primer and Mg2+, each at four levels. The optimal SRAP-PCR system for Lonicera caerulea L. was 20 ktL SRAP-PCR amplification reaction solution containing 2.0 μL 10×PCR buffer, 1.0 U Taq DNA polymerase, 30 ng template DNA, 0.2 mmol·L-1 dNTPs, 2.0 mmol·L-1 Mg2+ and 0.2μmol·L-1 primer. The suitable amplification procedure consisted of an initial denaturation at 94℃ for 5 min; denaturation at 94℃ for 1 min, annealing at 35℃ for 1 rain, extension at 72℃ for 90 s and in total five cycles; denaturation at 94℃ for 1 min, annealing at 50℃ for 1 min, extension at 72℃ for 90 s and in total 35 cycles; extension at 72℃ for 8 rain; preservation at 4℃. The procedures and systems could meet the demand for SRAP amplification of Lonicera caerulea L. and would play an important role in Lonicera caerulea L. germplasm identification and genetic diversity analysis.

  19. Polymerase Chain Reaction (PCR)-based methods for detection and identification of mycotoxigenic Penicillium species using conserved genes

    Science.gov (United States)

    Polymerase chain reaction amplification of conserved genes and sequence analysis provides a very powerful tool for the identification of toxigenic as well as non-toxigenic Penicillium species. Sequences are obtained by amplification of the gene fragment, sequencing via capillary electrophoresis of d...

  20. 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.