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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. Enzymatic Synthesis of Modified Oligonucleotides by PEAR Using Phusion and KOD DNA Polymerases

    OpenAIRE

    Wang, Xuxiang; Zhang, Jianye; Li, Yingjia; Chen, Gang; Wang, Xiaolong

    2015-01-01

    Antisense synthetic oligonucleotides have been developed as potential gene-targeted therapeutics. We previously reported polymerase–endonuclease amplification reaction (PEAR) for amplification of natural and 5′-O-(1-thiotriphosphate) (S)-modified oligonucleotides. Here, we extended the PEAR technique for enzymatic preparation of 2′-deoxy-2′-fluoro-(2′-F) and 2′-F/S double-modified oligonucleotides. The result showed that KOD and Phusion DNA polymerase could synthesize oligonucleotides with on...

  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. Enzymatic synthesis of modified oligonucleotides by PEAR using Phusion and KOD DNA polymerases.

    Science.gov (United States)

    Wang, Xuxiang; Zhang, Jianye; Li, Yingjia; Chen, Gang; Wang, Xiaolong

    2015-02-01

    Antisense synthetic oligonucleotides have been developed as potential gene-targeted therapeutics. We previously reported polymerase-endonuclease amplification reaction (PEAR) for amplification of natural and 5'-O-(1-thiotriphosphate) (S)-modified oligonucleotides. Here, we extended the PEAR technique for enzymatic preparation of 2'-deoxy-2'-fluoro-(2'-F) and 2'-F/S double-modified oligonucleotides. The result showed that KOD and Phusion DNA polymerase could synthesize oligonucleotides with one or two modified nucleotides, and KOD DNA polymerase is more suitable than Phusion DNA polymerase for PEAR amplification of 2'-F and 2'-F/S double modified oligonucleotides. The composition of PEAR products were analyzed by electrospray ionization liquid chromatography mass spectrometry (ESI/LC/MS) detection and showed that the sequence of the PEAR products are maintained at an extremely high accuracy (>99.9%), and after digestion the area percent of full-length modified oligonucleotides reaches 89.24%. PEAR is suitable for synthesis of modified oligonucleotides efficiently and with high purity. PMID:25517220

  5. Rapid Diagnosis of Extrapulmonary Tuberculosis by Ligase Chain Reaction Amplification

    OpenAIRE

    Gamboa, Fredy; Dominguez, José; Padilla, Eduardo; Manterola, José M.; Gazapo, Elena; Lonca, Joan; Matas, Lurdes; Hernandez, Agueda; Cardona, Pere Joan; Ausina, Vicente

    1998-01-01

    A rapid amplification-based test for the diagnosis of extrapulmonary tuberculosis, the LCx Mycobacterium tuberculosis Assay from Abbott Laboratories, was evaluated. Results from the LCx M. tuberculosis Assay were compared with those from culture and the final clinical diagnosis for each patient. A total of 526 nonrespiratory specimens from 492 patients were tested. The specimens included urine; feces; lymph node exudates; pleural, cerebrospinal, articular, and ascitic fluids; tissue biopsies;...

  6. Thermostable Mismatch-Recognizing Protein MutS Suppresses Nonspecific Amplification during Polymerase Chain Reaction (PCR

    Directory of Open Access Journals (Sweden)

    Seiki Kuramitsu

    2013-03-01

    Full Text Available Polymerase chain reaction (PCR-related technologies are hampered mainly by two types of error: nonspecific amplification and DNA polymerase-generated mutations. Here, we report that both errors can be suppressed by the addition of a DNA mismatch-recognizing protein, MutS, from a thermophilic bacterium. Although it had been expected that MutS has a potential to suppress polymerase-generated mutations, we unexpectedly found that it also reduced nonspecific amplification. On the basis of this finding, we propose that MutS binds a mismatched primer-template complex, thereby preventing the approach of DNA polymerase to the 3' end of the primer. Our simple methodology improves the efficiency and accuracy of DNA amplification and should therefore benefit various PCR-based applications, ranging from basic biological research to applied medical science.

  7. Series DNA Amplification Using the Continuous-Flow Polymerase Chain Reaction Chip

    Science.gov (United States)

    Joung, Seung-Ryong; Kang, Chi Jung; Kim, Yong-Sang

    2008-02-01

    We proposed a continuous-flow polymerase chain reaction (PCR) chip that can be used for series DNA amplification. The continuous-flow PCR chip has several advantages such as fast thermal cycling, series of amplifications, cost-effective fabrication, portability, and fluorescence detection. The continuous-flow PCR chip is composed of two parts namely poly(dimethylsiloxane) (PDMS) microchannel for sample injection and indium-tin-oxide (ITO) heater/glass chip for thermal cycling. The fabricated microchannel width and depth are 250 and 200 µm, respectively. Also, the total working length of the PDMS microchannel is 1340 mm which is equivalent for 20 cycles of amplification. A 2:2:3 microchannel length ratio for three different temperature zones namely denaturation, annealing, and extension was assigned, respectively. Upon the operation of the fabricated continuous-flow PCR chip, the amplification of plasmid DNA pKS-GFP with 720 base pairs and PG-noswsi with 300 base pairs were found successfully with a total reaction time of 15 min.

  8. Sequencing of simple sequence repeat anchored polymerase chain reaction amplification products of Biomphalaria glabrata

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    Caldeira Roberta L

    2002-01-01

    Full Text Available Simple sequence repeat anchored polymerase chain reaction amplification (SSR-PCR is a genetic typing technique based on primers anchored at the 5' or 3' ends of microsatellites, at high primer annealing temperatures. This technique has already been used in studies of genetic variability of several organisms, using different primer designs. In order to conduct a detailed study of the SSR-PCR genomic targets, we cloned and sequenced 20 unique amplification products of two commonly used primers, CAA(CT6 and (CA8RY, using Biomphalaria glabrata genomic DNA as template. The sequences obtained were novel B. glabrata genomic sequences. It was observed that 15 clones contained microsatellites between priming sites. Out of 40 clones, seven contained complex sequence repetitions. One of the repeats that appeared in six of the amplified fragments generated a single band in Southern analysis, indicating that the sequence was not widespread in the genome. Most of the annealing sites for the CAA(CT6 primer contained only the six repeats found within the primer sequence. In conclusion, SSR-PCR is a useful genotyping technique. However, the premise of the SSR-PCR technique, verified with the CAA(CT6 primer, could not be supported since the amplification products did not result necessarily from microsatellite loci amplification.

  9. Micromachined polymerase chain reaction system for multiple DNA amplification of upper respiratory tract infectious diseases.

    Science.gov (United States)

    Liao, Chia-Sheng; Lee, Gwo-Bin; Wu, Jiunn-Jong; Chang, Chih-Ching; Hsieh, Tsung-Min; Huang, Fu-Chun; Luo, Ching-Hsing

    2005-01-15

    This paper presents a micro polymerase chain reaction (PCR) chip for the DNA-based diagnosis of microorganism genes and the detection of their corresponding antibiotic-resistant genes. The micro PCR chip comprises cheap biocompatible soda-lime glass substrates with integrated thin-film platinum resistors as heating/sensing elements, and is fabricated using micro-electro-mechanical-system (MEMS) techniques in a reliable batch-fabrication process. The heating and temperature sensing elements are made of the same material and are located inside the reaction chamber in order to ensure a uniform temperature distribution. This study performs the detection of several genes associated with upper respiratory tract infection microorganisms, i.e. Streptococcus pneumoniae, Haemopilus influenze, Staphylococcu aureus, Streptococcus pyogenes, and Neisseria meningitides, together with their corresponding antibiotic-resistant genes. The lower thermal inertia of the proposed micro PCR chip relative to conventional bench-top PCR systems enables a more rapid detection operation with reduced sample and reagent consumption. The experimental data reveal that the high heating and cooling rates of the system (20 and 10 degrees C/s, respectively) permit successful DNA amplification within 15 min. The micro PCR chip is also capable of performing multiple DNA amplification, i.e. the simultaneous duplication of multiple genes under different conditions in separate reaction wells. Compared with the large-scale PCR system, it is greatly advantageous for fast diagnosis of multiple infectious diseases. Multiplex PCR amplification of two DNA segments in the same well is also feasible using the proposed micro device. The developed micro PCR chip provides a crucial tool for genetic analysis, molecular biology, infectious disease detection, and many other biomedical applications. PMID:15590288

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

    Science.gov (United States)

    Liu, Changchun; Sadik, Mohamed M.; Mauk, Michael G.; Edelstein, Paul H.; Bushman, Frederic D.; Gross, Robert; Bau, Haim H.

    2014-01-01

    Real-time amplification and quantification of specific nucleic acid sequences plays a major role in medical and biotechnological applications. In the case of infectious diseases, such as HIV, quantification of the pathogen-load in patient specimens is critical to assess disease progression and effectiveness of drug therapy. Typically, nucleic acid quantification requires expensive instruments, such as real-time PCR machines, which are not appropriate for on-site use and for low-resource settings. This paper describes a simple, low-cost, reaction-diffusion 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. The method was tested for HIV viral load monitoring and performed on par with conventional benchtop methods. The proposed method is suitable for nucleic acid quantification at point of care, compatible with multiplexing and high-throughput processing, and can function instrument-free. PMID:25477046

  11. 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.

  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. Diagnosis of Streptococcus pneumoniae meningitis by polymerase chain reaction amplification of the gene for pneumolysin

    Directory of Open Access Journals (Sweden)

    Juliana de A Matos

    2006-08-01

    Full Text Available Diagnosis of bacterial meningitis has long been based on classical methods of Gram stain, serological tests, and culture of cerebrospinal fluid (CSF. The performance of these methods, especially culture and direct smear, is thwarted by failure to detect bacteria following administration of antimicrobial agents and reluctance to performance lumbar punctures at admission. Indeed, patients with meningitis frequently receive antibiotics orally or by injection before the diagnosis is suspected or established. Thus an alternative method has become necessary to help clinicians and epidemiologists to management and control of bacterial meningitis. We evaluate the application of a polymerase chain reaction-based (PCR assay for amplification of pneumolysin gene (ply to diagnosis of Streptococcus pneumoniae meningitis. The PCR assay sensitivity for CSF was 96% (95% confidence interval, CI, 90-99% compared to a sensitivity of 59% for culture (95% CI 49-69%, 66% for Gram stain (95% CI 56-74%, and 78% for latex agglutination test (95% CI 69-86%; PCR specificity was 100% (95% CI 83-100%. PCR results were available within 4 h of the start of the assay. This molecular approach proved to be reliable and useful to identify this bacterium compared with other classical laboratory methods for identification of bacterial meningitis pathogens.

  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. G-quadruplex based two-stage isothermal exponential amplification reaction for label-free DNA colorimetric detection.

    Science.gov (United States)

    Nie, Ji; Zhang, De-Wen; Tie, Cai; Zhou, Ying-Lin; Zhang, Xin-Xiang

    2014-06-15

    A novel G-quadruplex based two-stage isothermal exponential amplification reaction (GQ-EXPAR) was developed for label-free DNA colorimetric detection in this work. The exponential amplified trigger DNA in the first stage can convert into G-quadruplex sequence EAD2 by a linear amplification circuit in the second stage. Created EAD2 can form G-quadruplex/hemin DNAzyme to act as a direct signal readout element. The GQ-EXPAR combines the exponential amplification of DNA sequence and the peroxidase-mimicking DNAzyme induced signal amplification, which achieves tandem dual-amplification. Taking advantages of isothermal incubation, this label-free homogeneous assay obviates the need of thermal cycling . As no complex synthesis or extra downstream operation is needed, the whole easy handling procedure can be finished in no more than 1h. This assay allows the sensing of the model DNA with the limit of detection to be 2.5pM. Moreover, it demonstrates good discrimination of mismatched sequences. The strategy has also been successfully implemented to sensitively detect Tay-Sachs genetic disorder mutant. PMID:24508547

  16. 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

  17. 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.

  18. Fragrant pear sexuality recognition with machine vision

    Science.gov (United States)

    Ma, Benxue; Ying, Yibin

    2006-10-01

    In this research, a method to identify Kuler fragrant pear's sexuality with machine vision was developed. Kuler fragrant pear has male pear and female pear. They have an obvious difference in favor. To detect the sexuality of Kuler fragrant pear, images of fragrant pear were acquired by CCD color camera. Before feature extraction, some preprocessing is conducted on the acquired images to remove noise and unnecessary contents. Color feature, perimeter feature and area feature of fragrant pear bottom image were extracted by digital image processing technique. And the fragrant pear sexuality was determined by complexity obtained from perimeter and area. In this research, using 128 Kurle fragrant pears as samples, good recognition rate between the male pear and the female pear was obtained for Kurle pear's sexuality detection (82.8%). Result shows this method could detect male pear and female pear with a good accuracy.

  19. Topics in free radical-mediated DNA damage: purines and damage amplification - superoxic reactions - bleomycin, the incomplete radiomimetic

    International Nuclear Information System (INIS)

    Only a small percentage of the DNA damage set by ionizing radiation in the living cell manifests itself as lethal. It is now increasingly accepted that clustered lesions may constitute the kind of damage that the repair enzymes cannot adequately deal with. The question is raised as to whether damage amplification reactions (radical transfer reactions) may contribute to these clustered lesions, and examples of such damage amplification reactions are given. In one example a purine is involved. With 2'-deoxy adenosine and 2'-deoxy guanosine it is shown that these purine nucleosides undergo unexpected radical reactions. Evidence for the radical transfer from the purine to the sugar moiety is provided by the formation of the 5'-aldehydes. These products have been assayed with 2-thiobarbituric acid (TBA), a reagent commonly applied to the detection of malonaldehyde. TBA-reactive material has also been assayed in γ-irradiated DNA, about one-third of this is free malonaldehyde, while the major part of the TBA-reactive material remains bound to the DNA. In contrast, bleomycin-treated DNA yields practically no free malonaldehyde, and the major TBA-reactive products are identified as the thymine and adenine base propenals. (Author)

  20. 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.

  1. 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.

  2. The impact of social amplification and attenuation of risk and the public reaction to mad cow disease in Canada.

    Science.gov (United States)

    Lewis, Roxanne E; Tyshenko, Michael G

    2009-05-01

    Following the detection of bovine spongiform encephalopathy (BSE) in Canada, and subsequently in the United States, confidence in the safety of beef products remained high. Consumers actually increased their consumption of beef slightly after the news of an increased risk from mad cow disease, which has been interpreted as public support for beef farmers and confidence in government regulators. The Canadian public showed a markedly different reaction to the news of domestic BSE than the furious and panicked responses observed in the United Kingdom, Germany, and Japan. Using the social amplification of risk framework, we show that, while other countries displayed social amplification of risk, Canada experienced a social attenuation of risk. The attenuated reaction in Canada toward mad cow disease and increased human health risks from variant Creutzfeldt-Jakob disease (vCJD) was due to the social context at the time when BSE was discovered domestically. Mortality, morbidity, and psychosocial impacts resulting from other major events such as severe acute respiratory syndrome (SARS), West Nile virus (WNV), and the U.S.-Iraq war made the theoretical risks of BSE and vCJD a lower priority, reducing its concern as a risk issue. PMID:19192234

  3. 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.

  4. Species identification of Crassostrea and Ostrea oysters by polymerase chain reaction amplification of the 5S rRNA gene.

    Science.gov (United States)

    Cross, Ismael; Rebordinos, Laureana; Diaz, Edgardo

    2006-01-01

    A specific multiplex polymerase chain reaction (PCR) was developed for the identification of Crassostrea angulata, C. gigas, Ostrea edulis, and O. stentina oyster species. Universal primers were used for the amplification of complete repetition units of 5S rDNA in each of the 4 species. The alignment of the obtained sequences was the basis for the specific design of species-specific primers (ED1, ED2, ST1, ST2, CR1, and CR2) located in the nontranscribed spacer regions. The different sizes of the species-specific amplicons, separated by agarose gel electrophoresis, allowed identification of Crassostrea and Ostrea species. A multiplex PCR with a set of the 6 designed primers showed that they did not interfere with each other and bound specifically to the DNA target. This genetic marker can be very useful for traceability of the species, application in the management of oyster cultures, and conservation of the genetic resources of the species. PMID:16512239

  5. A sensitive electrochemiluminescent aptasensor based on perylene derivatives as a novel co-reaction accelerator for signal amplification.

    Science.gov (United States)

    Yu, Yan-Qing; Zhang, Hai-Yu; Chai, Ya-Qin; Yuan, Ruo; Zhuo, Ying

    2016-11-15

    Herein, a novel signal amplification strategy was designed using the perylene derivative as the co-reaction accelerator toward graphene-CdTe quantum dots (G-CdTe)/S2O8(2-) system to construct a highly sensitive electrochemiluminescent (ECL) aptasensor for thrombin (TB) detection. Firstly, the G-CdTe nanocomposites were prepared by one-step method of in situ generating CdTe quantum dots onto the surface of the graphene oxide by using 3-mercaptopropionic acid as the CdTe QDs stabilizer. Then, a kind of perylene derivative (PTC-Lys), was synthesized by covalently binding L-lysine to 3,4,9,10-perylenetetracarboxylic acid, which was further immobilized onto the G-CdTe by the π-π* stacking and cross-linked the detection thrombin aptamer (TBA II) to obtain the TBA II/PTC-Lys/G-CdTe signal probes. It is worth pointing out that PTC-Lys acting as an efficient co-reaction accelerator interacted with the co-reactant of S2O8(2-) rather than G-CdTe to promote the more oxidant mediators of SO4(•-), which could further react with G-CdTe to produce excited state species G-CdTe* for emitting light. Compared with the G-CdTe/S2O8(2-) ECL system, our proposed strategy with the introduction of co-reaction accelerator of PTC-Lys exhibited ultra-high sensitivity to quantify the concentration of TB from 1.0×10(-7)nM to 10nM with a detection limit of 34aM. PMID:27148827

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2008-06-15

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

  7. 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.

  8. 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.

  9. Trials to control European pear sucker

    OpenAIRE

    Scheer, Christian; Trautmann, Martin

    2006-01-01

    European pear sucker causes financial penalties in many pear orchards. Honey dew exudations reduce quality. Another problem is the transmission of pear decline. In years of high infestation chemical treatments are complicated. Neither cutisan- nor sugar-treatments showed an appreciable reduction of the pest in the year 2005.

  10. 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.

  11. Optimisation of an asymmetric polymerase chain reaction assay for the amplification of single-stranded DNA from Wuchereria bancrofti for electrochemical detection

    OpenAIRE

    Vasuki Venkatesan; Sugeerappa Laxmanappa Hoti; Nagalakshmi Kamaraj; Somnath Ghosh; Kaushik Rajaram

    2013-01-01

    Single-stranded DNA (ssDNA) is a prerequisite for electrochemical sensor-based detection of parasite DNA and other diagnostic applications. To achieve this detection, an asymmetric polymerase chain reaction method was optimised. This method facilitates amplification of ssDNA from the human lymphatic filarial parasite Wuchereria bancrofti. This procedure produced ssDNA fragments of 188 bp in a single step when primer pairs (forward and reverse) were used at a 100:1 molar ratio in the presence ...

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. Comparative study of sensitivity, linearity, and resistance to inhibition of digital and nondigital polymerase chain reaction and loop mediated isothermal amplification assays for quantification of human cytomegalovirus.

    Science.gov (United States)

    Nixon, Gavin; Garson, Jeremy A; Grant, Paul; Nastouli, Eleni; Foy, Carole A; Huggett, Jim F

    2014-05-01

    Performing nucleic acid amplification techniques (NAATs) in digital format using limiting dilution provides potential advantages that have recently been demonstrated with digital polymerase chain reaction (dPCR). Key benefits that have been claimed are the ability to quantify nucleic acids without the need of an external calibrator and a greater resistance to inhibitors than real-time quantitative PCR (qPCR). In this study, we evaluated the performance of four NAATs, qPCR, dPCR, real-time quantitative loop mediated isothermal amplification (qLAMP), and digital LAMP (dLAMP), for the detection and quantification of human cytomegalovirus (hCMV). We used various DNA templates and inhibitors to compare the performance of these methods using a conventional real-time thermocycler platform (Bio-Rad CFX96) and a chip based digital platform (Fluidigm Biomark 12.765 Digital Array). dPCR performed well and demonstrated greater resistance to inhibitors than the other methods although this resistance did not apply equally to all inhibitors tested. dLAMP was found to be less sensitive than dPCR, but its quantitative performance was better than qLAMP, the latter being unable to quantify below 1000 copies. dLAMP was also more resistant to inhibitors than qLAMP. Unlike qPCR, both digital methods were able to quantify viral genomes without requiring a calibrator; however, neither can currently compete with the large reaction volumes, and thus the greater absolute sensitivity, of qPCR. With the introduction of digital instrumentation that will enable larger reaction volumes, digital amplification methods such as those evaluated in this study could potentially offer a robust alternative to qPCR for nucleic acid quantification. PMID:24684191

  17. TPR-MET oncogenic rearrangement: Detection by polymerase chain reaction amplification of the transcript and expression in human tumor cells lines

    International Nuclear Information System (INIS)

    Activation of the MET protooncogene by a rearrangement involving the fusion of TPR and MET specific gene sequences has been observed in a human osteosarcoma cell line (HOS) treated in vitro with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). No information has been available about the possible occurrence of this rearrangement in human tumors. To facilitate rapid screening of human cell lines and tumor samples for this specific gene rearrangement; the authors developed a sensitive detection method based on polymerase chain reaction (PCR) amplification of TPR-MET mRNA. cDNA was generated from cellular transcripts by using one of the PCR primers, which was then used as a template for PCR amplification of a 205-base-pair region carrying the breakpoint. An end-labeled internal probe was hybridized in solution to an aliquot of the PCR product for detecting amplification. Cells could be directly screened by the assay without prior isolation of RNA. A 205-base-pair DNA fragment characteristic of the TRP-MET rearrangement was detected in cell lines previously known to contain this altered sequence. The rearrangement was also detected at very low levels in the parental (nontransformed) cell line, HOS TE-85. A preliminary survey of cell lines derived from a variety of human tumors indicates that TPR-MET rearrangement occurred and was expressed at very low frequencies by cells from 7 of 14 tumors of nonhematopoietic origin

  18. Videodermatoscopy of pearly penile papules. Case reports

    Directory of Open Access Journals (Sweden)

    Hristo Petrov Dobrev

    2015-01-01

    Full Text Available Pearly penile papules are harmless dermatological condition characterized by small dome-shaped to filiform skin-colored papules located in one to several rows along the sulcus or corona of the glans penis. We describe six male patients with pearly penile papules whose diagnosis was assisted by means of videodermatoscopy. All lesions showed a characteristic whitish pink cobblestone appearance arranged in one or several rows. Each papule contained central dotted or commalike vessels and was surrounded by crescent-shaped whitish structures. Videodermatoscopy may assist the diagnosis of pearly penile papules and successfully differentiate them from genital warts and molluscum contagiosum.

  19. 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.

  20. 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.

  1. Isoform identification, recombinant production and characterization of the allergen lipid transfer protein 1 from pear (Pyr c 3).

    Science.gov (United States)

    Ramazzina, Ileana; Amato, Stefano; Passera, Elisabetta; Sforza, Stefano; Mistrello, Gianni; Berni, Rodolfo; Folli, Claudia

    2012-01-10

    Non-specific lipid transfer proteins belonging to LTP1 family represent the most important allergens for non pollen-related allergies to Rosaceae fruits in the Mediterranean area. Peach LTP1 (Pru p 3) is a major allergen and is considered the prototypic allergenic LTP. On the contrary, pear allergy without pollinosis seems to be under-reported when compared to other Rosaceae fruits suggesting that the as-yet-uncharacterized pear LTP1 (Pyr c 3) has in vivo a low allergenicity. We report here on the identification of four cDNAs encoding for LTP1 in pear fruits. The two isoforms exhibiting amino acid sequences most similar to those of peach and apple homologues were obtained as recombinant proteins. Such isoforms exhibited CD spectra and lipid binding ability typical of LTP1 family. Moreover, pear LTP1 mRNA was mainly found in the peel, as previously shown for other Rosaceae fruits. By means of IgE ELISA assays a considerable immunoreactivity of these proteins to LTP-sensitive patient sera was detected, even though allergic reactions after ingestion of pear were not reported in the clinical history of the patients. Finally, the abundance of LTP1 in protein extracts from pear peel, in which LTP1 from Rosaceae fruits is mainly confined, was estimated to be much lower as compared to peach peel. Our data suggest that the two isoforms of pear LTP1 characterized in this study possess biochemical features and IgE-binding ability similar to allergenic LTPs. Their low concentrations in pear might be the cause of the low frequency of LTP-mediated pear allergy. PMID:22015956

  2. Oligosaccharide formation during commercial pear juice processing.

    Science.gov (United States)

    Willems, Jamie L; Low, Nicholas H

    2016-08-01

    The effect of enzyme treatment and processing on the oligosaccharide profile of commercial pear juice samples was examined by high performance anion exchange chromatography with pulsed amperometric detection and capillary gas chromatography with flame ionization detection. Industrial samples representing the major stages of processing produced with various commercial enzyme preparations were studied. Through the use of commercially available standards and laboratory scale enzymatic hydrolysis of pectin, starch and xyloglucan; galacturonic acid oligomers, glucose oligomers (e.g., maltose and cellotriose) and isoprimeverose were identified as being formed during pear juice production. It was found that the majority of polysaccharide hydrolysis and oligosaccharide formation occurred during enzymatic treatment at the pear mashing stage and that the remaining processing steps had minimal impact on the carbohydrate-based chromatographic profile of pear juice. Also, all commercial enzyme preparations and conditions (time and temperature) studied produced similar carbohydrate-based chromatographic profiles. PMID:26988479

  3. 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.

  4. Videodermatoscopy of pearly penile papules. Case reports

    OpenAIRE

    Hristo Petrov Dobrev; Desislava Georgieva Nocheva

    2015-01-01

    Pearly penile papules are harmless dermatological condition characterized by small dome-shaped to filiform skin-colored papules located in one to several rows along the sulcus or corona of the glans penis. We describe six male patients with pearly penile papules whose diagnosis was assisted by means of videodermatoscopy. All lesions showed a characteristic whitish pink cobblestone appearance arranged in one or several rows. Each papule contained central dotted or commalike vessels...

  5. 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。

  6. 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.

  7. 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…

  8. 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

  9. 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.

  10. Polyphenolic Profile of Pear Leaves with Different Resistance to Pear Psylla (Cacopsylla pyri).

    Science.gov (United States)

    Fotirić Akšić, Milica M; Dabić, Dragana Č; Gašić, Uroš M; Zec, Gordan N; Vulić, Todor B; Tešić, Živoslav Lj; Natić, Maja M

    2015-09-01

    The European pear psylla, Cacopsylla pyri L. (Hemiptera: Psyllidae), is one of the most serious arthropod pests of pear. Since proper control of this pest is essential, better understanding of the complex plant-pest relationship is mandatory. This research deals with constitutive polyphenolic profiles in leaves of 22 pear cultivars of diverse origin (P. communis, P. pyrifolia, and P. pyrifolia × P. communis) and different resistance to psylla. The study was designed to show which differences in the polyphenolic profile of leaves from resistant and susceptible pear cultivars could be utilized as information in subsequent breeding programs. The results demonstrated that the leaves of Oriental pear cultivars contained much higher amounts of p-hydroxybenzoic acid, ferulic acid, aesculin, and naringin, that, together with detected 3-O-(6″-O-p-coumaroyl)-hexoside, apigenin, apigenin 7-O-rutinoside, and hispidulin, indicated a clear difference between the species and might represent phenolics responsible for psylla resistance. PMID:26278376

  11. 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.

  12. 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...

  13. 76 FR 78168 - Importation of Chinese Sand Pears From China

    Science.gov (United States)

    2011-12-16

    ... of Chinese Sand Pears From China AGENCY: Animal and Plant Health Inspection Service, USDA. ACTION... would set forth shipping requirements for sand pears from China. It would require sealed containers of... importation of Chinese sand pears (Pyrus pyrifolia) from China into the United States. As a condition of...

  14. Optimisation of an asymmetric polymerase chain reaction assay for the amplification of single-stranded DNA from Wuchereria bancrofti for electrochemical detection

    Directory of Open Access Journals (Sweden)

    Vasuki Venkatesan

    2013-09-01

    Full Text Available Single-stranded DNA (ssDNA is a prerequisite for electrochemical sensor-based detection of parasite DNA and other diagnostic applications. To achieve this detection, an asymmetric polymerase chain reaction method was optimised. This method facilitates amplification of ssDNA from the human lymphatic filarial parasite Wuchereria bancrofti. This procedure produced ssDNA fragments of 188 bp in a single step when primer pairs (forward and reverse were used at a 100:1 molar ratio in the presence of double-stranded template DNA. The ssDNA thus produced was suitable for immobilisation as probe onto the surface of an Indium tin oxide electrode and hybridisation in a system for sequence-specific electrochemical detection of W. bancrofti. The hybridisation of the ssDNA probe and target ssDNA led to considerable decreases in both the anodic and the cathodic currents of the system's redox couple compared with the unhybridised DNA and could be detected via cyclic voltammetry. This method is reproducible and avoids many of the difficulties encountered by conventional methods of filarial parasite DNA detection; thus, it has potential in xenomonitoring.

  15. Optimisation of an asymmetric polymerase chain reaction assay for the amplification of single-stranded DNA from Wuchereria bancrofti for electrochemical detection.

    Science.gov (United States)

    Venkatesan, Vasuki; Hoti, Sugeerappa Laxmanappa; Kamaraj, Nagalakshmi; Ghosh, Somnath; Rajaram, Kaushik

    2013-09-01

    Single-stranded DNA (ssDNA) is a prerequisite for electrochemical sensor-based detection of parasite DNA and other diagnostic applications. To achieve this detection, an asymmetric polymerase chain reaction method was optimised. This method facilitates amplification of ssDNA from the human lymphatic filarial parasite Wuchereria bancrofti. This procedure produced ssDNA fragments of 188 bp in a single step when primer pairs (forward and reverse) were used at a 100:1 molar ratio in the presence of double-stranded template DNA. The ssDNA thus produced was suitable for immobilisation as probe onto the surface of an Indium tin oxide electrode and hybridisation in a system for sequence-specific electrochemical detection of W. bancrofti. The hybridisation of the ssDNA probe and target ssDNA led to considerable decreases in both the anodic and the cathodic currents of the system's redox couple compared with the unhybridised DNA and could be detected via cyclic voltammetry. This method is reproducible and avoids many of the difficulties encountered by conventional methods of filarial parasite DNA detection; thus, it has potential in xenomonitoring. PMID:24037206

  16. 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.

  17. 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.

  18. 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.

  19. 21 CFR 145.175 - Canned pears.

    Science.gov (United States)

    2010-04-01

    ... Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION CANNED FRUITS Requirements for Specific Standardized Canned Fruits § 145.175 Canned pears. (a...) Water. (b) Fruit juice(s) and water. (c) Fruit juice(s). (d) Clarified juice. Such packing media may...

  20. 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.

  1. 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. PMID:27498326

  2. 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...

  3. Study of the Drying Kinetics of Pears from cultivar D. Joaquina

    OpenAIRE

    Barroca, Maria João; Guiné, Raquel

    2013-01-01

    Drying is one of the most widely used methods for preserving foods allowing extending their shelf life by minimizing the moisture content in the food so that the deterioration reactions will not be able to occur. The objective of the present work was to determine the drying kinetics of pears from cultivar D. Joaquina, and then use the experimental data to model the drying kinetics by means of different thin layer equations found in the literature. For that t...

  4. Dynamics and control of DNA sequence amplification

    International Nuclear Information System (INIS)

    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

  5. 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.

  6. Revealed Comparative Advantage and Competitiveness in Pear

    OpenAIRE

    Jaime De Pablo Valenciano; Giancinti, Miguel A.; Juan Uribe

    2012-01-01

    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.

  7. 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.

  8. Market Quality of Pacific Northwest Pears

    OpenAIRE

    Gallardo, Rosa Karina; Kupferman, Eugene M.; Beaudry, Randolph M.; Blankenship, Sylvia M.; Mitcham, Elizabeth J; Watkins, Christopher B

    2011-01-01

    This study uses data collected from retail grocery chains during marketing season 2003-2004 to examine the external quality and price variations of Pacific Northwest pears. Quality refers to overall fruit appearance and presence of external disorders. Results from a bivariate probit model show that fruit weight and firmness had a positive effect on overall appearance. Results from a hedonic price model show that the recurrence of external disorders is not necessarily negatively correlated wit...

  9. 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

    DEFF Research Database (Denmark)

    Hillig, Thore; Thode, Jørgen; Breinholt, Ellen Marie;

    2012-01-01

    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....... 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...

  10. 77 FR 75007 - Importation of Sand Pears From China

    Science.gov (United States)

    2012-12-19

    ..., 2011, we published a proposed rule \\2\\ in the Federal Register (76 FR 78168- 78172, Docket No. APHIS... 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...

  11. Postharvest Decay of Apples and Pears in The Netherlands

    NARCIS (Netherlands)

    Wenneker, M.; Köhl, J.

    2014-01-01

    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 60% of fruit losses are recorded). Over 125 heavily affected lots of apples (mainly ‘Elstar’) and pears (mainly ‘Conference’)

  12. 77 FR 72245 - Pears Grown in Oregon and Washington; Committee Membership Reapportionment for Processed Pears

    Science.gov (United States)

    2012-12-05

    ... behalf. Thus, both statutes have small entity orientation and compatibility. There are approximately 1... order's information collection requirements have been previously approved by the Office of Management...; ] DEPARTMENT OF AGRICULTURE Agricultural Marketing Service 7 CFR Part 927 Pears Grown in Oregon and...

  13. 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...

  14. Hybrid Chirped Pulse Amplification

    Energy Technology Data Exchange (ETDEWEB)

    Jovanovic, I; Barty, C P J

    2002-05-07

    We present a novel chirped pulse amplification method which combines optical parametric amplification and laser amplification. We have demonstrated this hybrid CPA concept with a combination of beta-barium borate and Ti:sapphire. High-efficiency, multi-terawatt compatible amplification is achieved without gain narrowing and without electro-optic modulators using a simple commercial pump laser.

  15. Effects of fixative and fixation time on the extraction and polymerase chain reaction amplification of RNA from paraffin-embedded tissue. Comparison of two housekeeping gene mRNA controls.

    Science.gov (United States)

    Foss, R D; Guha-Thakurta, N; Conran, R M; Gutman, P

    1994-09-01

    A number of reports have indicated that RNA recovered from paraffin-embedded tissue can be used as a substrate in the polymerase chain reaction (PCR). Although it is established that RNA in paraffin-embedded tissue undergoes significant degradation, the specific contributions of different fixatives and fixation times to this degradation are not known. Mouse splenic tissue was harvested and fixed immediately for 2, 8, or 24 h in either formalin, Omnifix II, or Carnoy's fixative and then processed and embedded in paraffin. RNA was extracted from deparaffinized cubes of tissue using an adaptation of the technique described by Chomczynski and Sacchi. RNA was reverse transcribed using a random hexamer primed reaction. PCR amplification for cDNAs of the housekeeping genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and hypoxanthine phosphoribosyltransferase (HPRT) mRNAs was then performed. Although GAPDH amplification is used routinely on fresh and frozen tissues, we show that the presence of DNA contamination in the RNA preparations limits its usefulness in paraffin-embedded tissue. Amplifiable HPRT mRNA sequences were detected in nine of 12 samples fixed in Omnifix II, in four of 12 samples fixed in Carnoy's fixative, and in none of 12 formalin-fixed samples. Because of primer selection to preclude amplification of genomic HPRT, DNA contamination is not an issue when HPRT is amplified. Thus, HPRT represents the control system of choice for the evaluation of RNA in PET. The techniques described provide a rapid, uniform, and reproducible method of obtaining RNA from PET for molecular analysis, but they indicate limited utility for retrospective analysis of archival tissues. PMID:7981889

  16. Isothermal DNA amplification in vitro: the helicase-dependent amplification system.

    Science.gov (United States)

    Jeong, Yong-Joo; Park, Kkothanahreum; Kim, Dong-Eun

    2009-10-01

    Since the development of polymerase chain reaction, amplification of nucleic acids has emerged as an elemental tool for molecular biology, genomics, and biotechnology. Amplification methods often use temperature cycling to exponentially amplify nucleic acids; however, isothermal amplification methods have also been developed, which do not require heating the double-stranded nucleic acid to dissociate the synthesized products from templates. Among the several methods used for isothermal DNA amplification, the helicase-dependent amplification (HDA) is discussed in this review with an emphasis on the reconstituted DNA replication system. Since DNA helicase can unwind the double-stranded DNA without the need for heating, the HDA system provides a very useful tool to amplify DNA in vitro under isothermal conditions with a simplified reaction scheme. This review describes components and detailed aspects of current HDA systems using Escherichia coli UvrD helicase and T7 bacteriophage gp4 helicase with consideration of the processivity and efficiency of DNA amplification. PMID:19629390

  17. Label-free cascade amplification strategy for sensitive visual detection of thrombin based on target-triggered hybridization chain reaction-mediated in situ generation of DNAzymes and Pt nanochains.

    Science.gov (United States)

    Zhang, Ying; Ren, Wang; Luo, Hong Qun; Li, Nian Bing

    2016-06-15

    A new magnetic bead-based cascade amplification strategy for highly sensitive visual detection of proteins (thrombin as a model analyte) was developed by coupling target-triggered hybridization chain reaction (HCR) with the synergistic catalysis of DNA concatemer-mediated hemin/G-quadruplex DNAzymes and Pt nanozymes. Initially, the biotinylated primer DNA (P-DNA) was complementary with aptamer to form dsDNA which was further linked to streptavidin-coated magnetic bead (MB), thereby fabricating the expected MB-based aptasensor. In the presence of target TB, the aptamer was taken away from the aptasensor, and the free P-DNA immediately triggered HCR to spontaneously form DNA concatemer-directed nanochains with numerous DNAzymes and Pt nanoclusters (PtNCs) to achieve cascades signal amplification. The dual peroxidase mimetics catalyzed the H2O2-mediated oxidation of colorless 3,3',5,5'-tetramethylbenzidine (TMB) into the colored TMB oxides (oxTMB), causing intensified color change of the chromogenic solution for the highly sensitive naked-eye detection of as low as 100.0 pM TB. In this strategy, the employment of magnetic separation and exonuclease III (Exo III)-assisted digestion of residual dsDNA minimized the background noise and avoided the false positive results, greatly improving the detection accuracy and sensitivity with a low limit of detection (LOD=15.0 pM). The proposed visual platform has promise for detecting various types of proteins with careful DNA sequence designs. PMID:26878483

  18. Genomics of pear and other Rosaceae fruit trees.

    Science.gov (United States)

    Yamamoto, Toshiya; Terakami, Shingo

    2016-01-01

    The family Rosaceae includes many economically important fruit trees, such as pear, apple, peach, cherry, quince, apricot, plum, raspberry, and loquat. Over the past few years, whole-genome sequences have been released for Chinese pear, European pear, apple, peach, Japanese apricot, and strawberry. These sequences help us to conduct functional and comparative genomics studies and to develop new cultivars with desirable traits by marker-assisted selection in breeding programs. These genomics resources also allow identification of evolutionary relationships in Rosaceae, development of genome-wide SNP and SSR markers, and construction of reference genetic linkage maps, which are available through the Genome Database for the Rosaceae website. Here, we review the recent advances in genomics studies and their practical applications for Rosaceae fruit trees, particularly pear, apple, peach, and cherry. PMID:27069399

  19. 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.

  20. Properties of pears dried with different drying processes.

    OpenAIRE

    Guiné, Raquel; Barroca, Maria João; Lopes, Paulo; Silva, Vitor; Lima, Maria João; Ferreira, Dulcineia

    2010-01-01

    Pears of S. Bartolomeu variety have been used over the years in Portugal to produce a traditional dried pear using the cheapest practice with environmental friendly characteristics: open-sun drying. However, the obvious disadvantages concerning the drying efficiency and safety of the dried product have been an incentive to design or improve solar drying methods as an alternative to the traditional one. The present work has tested three types of drying processes...

  1. 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.

  2. Quadruple signal amplification strategy based on hybridization chain reaction and an immunoelectrode modified with graphene sheets, a hemin/G-quadruplex DNAzyme concatamer, and alcohol dehydrogenase: ultrasensitive determination of influenza virus subtype H7N9

    International Nuclear Information System (INIS)

    We report on a new amplification strategy for use in an immunoassay for influenza virus subtype H7N9. Graphene sheets were first placed on a glassy carbon electrode (GCE), and gold nanoparticles were then electrodeposited as a support for a layer of alcohol dehydrogenase (ADH) in a sol–gel containing thiol groups. Protein A was used to properly orientate immobilized antibody against H7N9 on the sol–gel, and this is shown to result in strongly improved specificity of the antigen-antibody binding. Thus, a sensitive and specific immunosensor was obtained in which a quadruple signal amplification strategy is employed, viz. (a) via the use of graphene sheets, (b) via a hybridization chain reaction, (c) the use of hemin/G-quadruplex DNAzyme concatamers, and (d) the use of ADH. The hemin/G-quadruplex is a typical DNAzyme, which simultaneously acts as NADH oxidase and HRP-mimicking DNAzyme. The hybridization chain reaction-based DNAzyme concatamers assembled on multi-walled carbon nanotubes (MWCNTs) and the ADH represent a triple electrocatalytic enzyme cascade system. Sandwich immunoreactions occurred between the capture antibody on the electrode and the secondary antibody labeled with MWCNTs. Positively charged Methylene Blue (MB) was then used as an intercalator to detect the DNAzyme concatamer formed. The differential pulse voltammetric signals for MB are related to the concentration of H7N9 in the range from 8 to 60 pg · mL−1, and the detection limit is 0.81 pg · mL−1 (at an S/N ratio of 3). This immunoassay is very sensitive, specific and robust. (author)

  3. 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.

  4. 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.

  5. 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

  6. 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

  7. Amplification-Free Detection of Circulating microRNA Biomarkers from Body Fluids Based on Fluorogenic Oligonucleotide-Templated Reaction between Engineered Peptide Nucleic Acid Probes: Application to Prostate Cancer Diagnosis.

    Science.gov (United States)

    Metcalf, Gavin A D; Shibakawa, Akifumi; Patel, Hinesh; Sita-Lumsden, Ailsa; Zivi, Andrea; Rama, Nona; Bevan, Charlotte L; Ladame, Sylvain

    2016-08-16

    Highly abundant in cells, microRNAs (or miRs) play a key role as regulators of gene expression. A proportion of them are also detectable in biofluids making them ideal noninvasive biomarkers for pathologies in which miR levels are aberrantly expressed, such as cancer. Peptide nucleic acids (PNAs) are engineered uncharged oligonucleotide analogues capable of hybridizing to complementary nucleic acids with high affinity and high specificity. Herein, novel PNA-based fluorogenic biosensors have been designed and synthesized that target miR biomarkers for prostate cancer (PCa). The sensing strategy is based on oligonucleotide-templated reactions where the only miR of interest serves as a matrix to catalyze an otherwise highly unfavorable fluorogenic reaction. Validated in vitro using synthetic RNAs, these newly developed biosensors were then shown to detect endogenous concentrations of miR in human blood samples without the need for any amplification step and with minimal sample processing. This low-cost, quantitative, and versatile sensing technology has been technically validated using gold-standard RT-qPCR. Compared to RT-qPCR however, this enzyme-free, isothermal blood test is amenable to incorporation into low-cost portable devices and could therefore be suitable for widespread public screening. PMID:27498854

  8. 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)

  9. 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.

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

    Science.gov (United States)

    2010-01-01

    ... apples or pears that are offered for entry into the United States. If inspection of the sample discloses... 7 Agriculture 5 2010-01-01 2010-01-01 false Apples and pears from Australia (including Tasmania... Fruits and Vegetables § 319.56-20 Apples and pears from Australia (including Tasmania) and New...

  11. 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....

  12. 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...

  13. 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.

  14. Two Pear Glutathione S-Transferases Genes Are Regulated during Fruit Development and Involved in Response to Salicylic Acid, Auxin, and Glucose Signaling

    OpenAIRE

    Hai-Yan Shi; Zheng-Hong Li; Yu-Xing Zhang; Liang Chen; Di-Ying Xiang; Yu-Feng Zhang

    2014-01-01

    Two genes encoding putative glutathione S-transferase proteins were isolated from pear (Pyrus pyrifolia) and designated PpGST1 and PpGST2. The deduced PpGST1 and PpGST2 proteins contain conserved Glutathione S-transferase N-terminal domain (GST_N) and Glutathione S-transferase, C-terminal domain (GST_C). Using PCR amplification technique, the genomic clones corresponding to PpGST1 and PpGST2 were isolated and shown to contain two introns and a singal intron respectively with typical GT/AG bou...

  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. 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.

  17. Antioxidant components and physico-chemical characteristics of jamun powder supplemented pear juice.

    Science.gov (United States)

    Kapoor, Swati; Ranote, Pushpinder Singh

    2016-05-01

    Studies were conducted to develop jamun powder supplemented pear juice. Two drying methods (Hot air cabinet drying and freeze drying) were used to prepare jamun pulp powder. Jamun powder was then blended with pear juice at 1, 2, 3, 4 and 5 % levels for preparation of jamun powder supplemented pear juice. Among the drying methods used, freeze dried powder retained better bioactive compounds and possessed higher antioxidant activity as compared to hot air dried jamun powder. Analysis of color properties (L*, a*, b*) revealed lower L*, b* values and higher a* values with progression of supplementation levels indicating decreased brightness of product. Pear juice supplemented with 4 % jamun powder received highest overall acceptability scores and was chosen for development of final product. Physico-chemical characteristics of control pear juice did not vary much from when compared to jamun powder supplemented pear juice. Bioactive components mainly total phenols enhanced (9.24 % higher) with addition of jamun powder in pear juice. Addition of anthocyanins from jamun powder to pear juice upon blending improved antioxidant activity of the final product. Supplemented pear juice had 18.13 % higher antioxidant activity than pear juice without supplementation. Storage period of 6 months resulted in significant (p < 0.05) decrease of bioactive compounds and antioxidant activity in jamun powder supplemented pear juice. PMID:27407197

  18. 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

    2015-01-01

    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. PMID:25993418

  19. 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.

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

    Directory of Open Access Journals (Sweden)

    Hai-Yan Shi

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

  1. 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. PMID:26152820

  2. 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.

  3. 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.

  4. Hypothetical pear production system innovation through promotion of soil biodiversity

    NARCIS (Netherlands)

    Maas, van der M.P.; Heijne, B.

    2014-01-01

    We developed a hypothetical view on a pear production system innovation through promotion of soil biodiversity. Two types of measures form the central idea of the innovation. The first type of measure is to replace part of the current 100% mineral fertilisation by compost plus targeted use of specia

  5. 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.

  6. 7 CFR 457.111 - Pear crop insurance provisions.

    Science.gov (United States)

    2010-01-01

    ... Provisions with (1) controlling (2), etc. 1. Definitions Direct marketing. Sale of the insured crop directly..., processor, shipper, or buyer. Examples of direct marketing include selling through an on-farm or roadside... alternating or mixed pattern. Marketable. Pear production acceptable for processing or other human...

  7. 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...

  8. 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. PMID:25384245

  9. Allele-specific DNA methylation reinforces PEAR1 enhancer activity.

    Science.gov (United States)

    Izzi, Benedetta; Pistoni, Mariaelena; Cludts, Katrien; Akkor, Pinar; Lambrechts, Diether; Verfaillie, Catherine; Verhamme, Peter; Freson, Kathleen; Hoylaerts, Marc F

    2016-08-18

    Genetic variation in the PEAR1 locus is linked to platelet reactivity and cardiovascular disease. The major G allele of rs12041331, an intronic cytosine guanine dinucleotide-single-nucleotide polymorphism (CpG-SNP), is associated with higher PEAR1 expression in platelets and endothelial cells than the minor A allele. The molecular mechanism underlying this difference remains elusive. We have characterized the histone modification profiles of the intronic region surrounding rs12041331 and identified H3K4Me1 enhancer-specific enrichment for the region that covers the CpG-SNP. Interestingly, methylation studies revealed that the CpG site is fully methylated in leukocytes of GG carriers. Nuclear protein extracts from megakaryocytes, endothelial cells, vs control HEK-293 cells show a 3-fold higher affinity for the methylated G allele compared with nonmethylated G or A alleles in a gel electrophoretic mobility shift assay. To understand the positive relationship between methylation and gene expression, we studied DNA methylation at 4 different loci of PEAR1 during in vitro megakaryopoiesis. During differentiation, the CpG-SNP remained fully methylated, while we observed rapid methylation increases at the CpG-island overlapping the first 5'-untranslated region exon, paralleling the increased PEAR1 expression. In the same region, A-allele carriers of rs12041331 showed significantly lower DNA methylation at CGI1 compared with GG homozygote. This CpG-island contains binding sites for the methylation-sensitive transcription factor CTCF, whose binding is known to play a role in enhancer activation and/or repression. In conclusion, we report the molecular characterization of the first platelet function-related CpG-SNP, a genetic predisposition that reinforces PEAR1 enhancer activity through allele-specific DNA methylation. PMID:27313330

  10. 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.

  11. 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.

  12. 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。

  13. Early amplification options.

    Science.gov (United States)

    Gabbard, Sandra Abbott; Schryer, Jennifer

    2003-01-01

    Children with permanent hearing loss have been remediated with hearing amplification devices for decades. The influx of young infants identified with hearing loss through successful newborn hearing screening programs has established a need for amplification resources for infants within the first six months of life. For the approximately two of every 1000 infants born who are identified with bilateral hearing loss [Mehl and Thomson, 1998, Pediatrics 101, p. e4], the use of amplification is commonly the first step in treating the sequella of their loss. The use of hearing aids, combined with early intervention, has been shown to significantly improve the speech and language skills of young children with hearing loss [Yoshinaga-Itano, 2000, Seminars in Hearing 21, p. 309]. Speech and language delays have contributed to compromised academic performance of school aged children with hearing loss [Johnson et al., 1997, Educational Audiology Handbook, Singular Publishing, San Diego]. Most hard-of-hearing and deaf children use hearing aids and other assistive listening devices every day throughout their lifetime and the life expectancy of a hearing aid is only five to eight years. The current challenge for pediatric audiologists is selecting and evaluating the available amplification to provide the best options for children and their families. Amplification technology has seen an explosion in growth the past few years and the options continue to expand rapidly. This article examines currently available amplification technology and reviews the selection criteria that may be used for infants and young children. Issues such as style, type, amplification features, signal processing strategies, and verification and validation tools are also discussed. PMID:14648816

  14. 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

  15. Quantum Feedback Amplification

    Science.gov (United States)

    Yamamoto, Naoki

    2016-04-01

    Quantum amplification is essential for various quantum technologies such as communication and weak-signal detection. However, its practical use is still limited due to inevitable device fragility that brings about distortion in the output signal or state. This paper presents a general theory that solves this critical issue. The key idea is simple and easy to implement: just a passive feedback of the amplifier's auxiliary mode, which is usually thrown away. In fact, this scheme makes the controlled amplifier significantly robust, and furthermore it realizes the minimum-noise amplification even under realistic imperfections. Hence, the presented theory enables the quantum amplification to be implemented at a practical level. Also, a nondegenerate parametric amplifier subjected to a special detuning is proposed to show that, additionally, it has a broadband nature.

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

    OpenAIRE

    Q Tri Ho; Pieter Verboven; Verlinden, Bert E.; Jeroen Lammertyn; Stefan Vandewalle; Nicolaï, Bart M.

    2008-01-01

    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...

  17. Cactus pear: a fruit of nutraceutical and functional importance

    OpenAIRE

    Piga, Antonio

    2004-01-01

    The constantly increasing demand for nutraceuticals is paralleled by a more pronounced request for natural ingredients and health-promoting foods. The multiple functional properties of cactus pear fit well this trend. Recent data revealed the high content of some chemical constituents, which can give added value to this fruit on a nutritional and technological functionality basis. High levels of betalains, taurine, calcium, magnesium, and antioxidants are noteworthy.

  18. Energy efficiency of organic pear production in greenhouses in China

    OpenAIRE

    Liu, Y.; Langer, Vibeke; Høgh-Jensen, Henning; Egelyng, Henrik

    2010-01-01

    The development of organic protected cultivation taking place in densely populated areas has raised the question whether it is an environmentally friendly production system. The present study investigated energy consumption of organic pear production in two production systems, namely in traditional Chinese solar greenhouse and in the open field. In both production systems, energy output/input ratio and energy productivity were used as indicators to determine the energy efficiency; yield, cost...

  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. PMID:25366556

  20. 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.

  1. 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.

  2. 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.

  3. 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

  4. Evaluation of antidiabetic properties of cactus pear seed oil in rats

    OpenAIRE

    Berraaouan, Ali; Ziyyat, Abderrahim; Mekhfi, Hassane; Legssyer, Abdelkhaleq; Sindic, Marianne; Aziz, Mohammed; Bnouham, Mohamed

    2014-01-01

    Cactus pear (Opuntia ficus-indica (L.) Mill. (Cactaceae)) is a medicinal plant widely used to treat diabetes. This work investigates the hypoglycemic and antihyperglycemic effect of cactus pear seed oil (CPSO), its mechanism of action, and any toxic effects. Peer reviewed

  5. A proposed mechanism behind the development of internal browning in pears (Pyrus communis cv Conference)

    NARCIS (Netherlands)

    Veltman, R.H.; Peppelenbos, H.W.

    2003-01-01

    Storage of pears under low oxygen levels (0.5-1.0 kPa) leads to decreased ascorbic acid and ATP levels, a lower ATP-production, and to internal browning, a storage disorder in pears. Addition of 5 kPa carbon dioxide to the storage atmosphere increased the severity of this disorder. Experiments showe

  6. 7 CFR 927.120 - Pears for charitable or byproduct purposes.

    Science.gov (United States)

    2010-01-01

    ... intended receiver and user of said pears showing, to the manager's satisfaction, that said pears actually... MARKETING SERVICE (Marketing Agreements and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE..., or quality regulations shall not be shipped or handled for consumption by any charitable...

  7. 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.

  8. 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.

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

    International Nuclear Information System (INIS)

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

  10. Flux amplification in SSPX

    Science.gov (United States)

    Lodestro, Lynda; Hooper, E. B.; Jayakumar, R. J.; Pearlstein, L. D.; Wood, R. D.; McLean, H. S.

    2007-11-01

    Flux amplification---the ratio of poloidal flux enclosed between the magnetic and geometric axes to that between the separatrix and the geometric axis---is a key measure of efficiency for edge-current-driven spheromaks. With the new, modular capacitor bank, permitting flexible programming of the gun current, studies of flux amplification under various drive scenarios can be performed. Analysis of recent results of pulsed operation with the new bank finds an efficiency ˜ 0.2, in selected shots, of the conversion of gun energy to confined magnetic energy during the pulses, and suggests a route toward sustained efficiency at 0.2. Results of experiments, a model calculation of field build-up, and NIMROD simulations exploring this newly suggested scenario will be presented.

  11. THE PEAR TREE RESPONSE TO PHOSPHORUS AND POTASSIUM FERTILIZATION

    Directory of Open Access Journals (Sweden)

    GUSTAVO BRUNETTO

    2015-06-01

    Full Text Available The aim of this study was to evaluate the response to phosphorus (P and potassium (K fertilization and to establish the critical levels of P and K in the soil and in the plant tissue in pear trees. Two experiments were conducted in São Joaquim (SC, Brazil. In experiment 1, the plants received annually the application of increasing rates of phosphate fertilizer (0, 40, 80, 120 and 160 kg P2O5 ha-1, while in experiment 2, increasing rates of potassium fertilizer (0, 40, 80, 120 and 160 kg K2O ha-1 were applied annually. In the two experiments, soil was collected annually from the 0-10, 10-20 and 0-20 cm layers, and the available P (experiment 1 and exchangeable K (experiment 2 content was analyzed. Whole leaves were collected annually, which were subjected to analysis of total P (experiment 1 and total K (experiment 2 content. The number and weight of the fruits per plant and fruit yield were evaluated. Application of P on the soil planted with pear trees increased the nutrient content in the soil and, in most crop seasons, in the whole leaf, but it did not affect the yield components and fruit yield. The application of K on the soil with pear trees increased the nutrient content in the soil and, in most of the crop seasons, in the whole leaf, but the potassium content in the whole leaf decreased in the crop season with greater fruit yield. The yield components and fruit yield were not affected by K fertilization.

  12. 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.

  13. Modelling of the thin layer drying kinetics of pears.

    OpenAIRE

    Guiné, Raquel; Barroca, Maria João; Lima, Maria João; Ferreira, Dulcienia

    2010-01-01

    Open-air sun drying has been traditionally used to dry grains, vegetables, fruits and other agricultural products. This is a common method of preserving foods and it is practiced until today in many countries where the climatic conditions are appropriate [1]. “Pera Passa de Viseu” denominates a traditional food product made after pears of the variety S. Bartolomeu using a traditional open-air sun drying [2]. Even though it is quite a cheap drying method, it has many important disadvantage...

  14. Construction Strategy for an Internal Amplification Control for Real-Time Diagnostic Assays Using Nucleic Acid Sequence-Based Amplification: Development and Clinical Application

    OpenAIRE

    Rodríguez-Lázaro, David; D'Agostino, Martin; Pla, Maria; Cook, Nigel

    2005-01-01

    An important analytical control in molecular amplification-based methods is an internal amplification control (IAC), which should be included in each reaction mixture. An IAC is a nontarget nucleic acid sequence which is coamplified simultaneously with the target sequence. With negative results for the target nucleic acid, the absence of an IAC signal indicates that amplification has failed. A general strategy for the construction of an IAC for inclusion in molecular beacon-based real-time nu...

  15. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and amplification refractory mutation system (ARMS) analyses of medicinally used Rheum species and their application for identification of Rhei Rhizoma.

    Science.gov (United States)

    Yang, Dong-Ye; Fushimi, Hirotoshi; Cai, Shao-Qing; Komatsu, Katsuko

    2004-05-01

    Previously, we have determined marker nucleotides on the chloroplast matK gene to identify Rheum palmatum, R. tanguticum and R. officinale used as Rhei Rhizoma officially. In the present study, we further developed a convenient and efficient identification method on the basis of marker nucleotides with Amplification Refractory Mutation System analysis. On the basis of the nucleotide substitutions at positions 367 and 937 among the three species on the matK gene, at each position two kinds of reverse primers with complementary 3'-terminal nucleotides were designed. Upon PCR amplification using three sets of primers and template DNA from each species, one or two fragments (202 bp or/and 770 bp) were detected. As the resultant three fragment profiles were species-specific, the procedure enabled us to classify the botanic origins of 22 drug samples of Rhei Rhizoma. PMID:15133241

  16. Coherent white light amplification

    Science.gov (United States)

    Jovanovic, Igor; Barty, Christopher P.

    2004-05-25

    A system for coherent simultaneous amplification of a broad spectral range of light that includes an optical parametric amplifier and a source of a seed pulse is described. A first angular dispersive element is operatively connected to the source of a seed pulse. A first imaging telescope is operatively connected to the first angular dispersive element and operatively connected to the optical parametric amplifier. A source of a pump pulse is operatively connected to the optical parametric amplifier. A second imaging telescope is operatively connected to the optical parametric amplifier and a second angular dispersive element is operatively connected to the second imaging telescope.

  17. 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•.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. Isothermal DNA amplification strategies for duplex microorganism detection.

    Science.gov (United States)

    Santiago-Felipe, Sara; Tortajada-Genaro, Luis Antonio; Morais, Sergi; Puchades, Rosa; Maquieira, Ángel

    2015-05-01

    A valid solution for micro-analytical systems is the selection of a compatible amplification reaction with a simple, highly-integrated efficient design that allows the detection of multiple genomic targets. Two approaches under isothermal conditions are presented: recombinase polymerase amplification (RPA) and multiple displacement amplification (MDA). Both methods were applied to a duplex assay specific for Salmonella spp. and Cronobacter spp., with excellent amplification yields (0.2-8.6 · 10(8) fold). The proposed approaches were successfully compared to conventional PCR and tested for the milk sample analysis as a microarray format on a compact disc (support and driver). Satisfactory results were obtained in terms of resistance to inhibition, selectivity, sensitivity (10(1)-10(2)CFU/mL) and reproducibility (below 12.5%). The methods studied are efficient and cost-effective, with a high potential to automate microorganisms detection by integrated analytical systems working at a constant low temperature.

  3. 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.

  4. Efficient Audio Power Amplification - Challenges

    DEFF Research Database (Denmark)

    Andersen, Michael Andreas E.

    2005-01-01

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

  5. 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)

  6. Internal browning disorder of eight pear cultivars affected by bioactive constituents and enzyme activity.

    Science.gov (United States)

    Koushesh Saba, Mahmoud; Moradi, Samira

    2016-08-15

    Internal browning (IB) is a disorder in pears that is frequently observed in some cultivars. The present research was carried out to study biochemical changes and IB disorder of pear fruit during storage and ripening. Eight pear cultivars harvested and stored at 1°C up to 90days. IB incidence, some bioactive compounds, polyphenol oxidase (PPO), peroxidase (POX), and superoxide dismutase (SOD) enzymes activities were measured during storage. IB increased during storage time but the susceptibility of cultivars was different. The ascorbic acid (AA), antioxidant capacity (AC) and SOD activity decreased while POX activity increased during storage but the rate of changes were different in studied cultivars. Total phenol (TP) and total flavonoid (TF) average content varied among pear cultivars and the highest TP and TF were observed in 'Bakhi' cultivars during storage. Fruit IB had positive correlation with the PPO activity, but negative correlation with TP, AC and AA. PMID:27006238

  7. Laser Induced Breakdown Spectroscopy of Prickly Pear's Spines and Glochids: A qualitative analysis

    International Nuclear Information System (INIS)

    A qualitative LIBS analysis of Prickly Pear is presented. The spectra for Q:Switch regime from cladode and spine are similar, while shows an intense electronic noise due the high absorption in spines for free-running regime

  8. Effect of Korean pear (Pyruspyrifolia cv. Shingo) juice on hangover severity following alcohol consumption.

    Science.gov (United States)

    Lee, Ho-Sun; Isse, Toyohi; Kawamoto, Toshihiro; Baik, Hyun Wook; Park, Jong Y; Yang, Mihi

    2013-08-01

    Korean pear has been used as a traditional prophylactic agent for alcohol hangover. However, its mechanism was not investigated in human yet. Therefore, we performed a randomized single blind crossover trial with 14 healthy young men to examine effects of Korean pear juice on alcohol hangover. All subjects consumed 540 ml of spirits (alcohol conc. 20.1 v/v%) after 30 min from the intervention, i.e. placebo or Korean pear juice treatment. Blood and urine specimens were collected in time-courses (9 time-points for 15 h after alcohol consumption). The total and average of hangover severity were alleviated to 16% and 21% by Korean pear juice at 15 h after the alcohol consumption, respectively (pshangover and its detoxification of alcohol seems to be modified by the genetic variation of ALDH2. PMID:23587660

  9. 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...

  10. 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.

  11. 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. PMID:26906904

  12. Draft Genome Sequence of Xylella fastidiosa Pear Leaf Scorch Strain in Taiwan.

    Science.gov (United States)

    Su, C-C; Deng, W-L; Jan, F-J; Chang, C-J; Huang, H; Chen, J

    2014-01-01

    The draft genome sequence of Xylella fastidiosa pear leaf scorch strain PLS229, isolated from the pear cultivar Hengshan (Pyrus pyrifolia) in Taiwan, is reported here. The bacterium has a genome size of 2,733,013 bp, with a G+C content of 53.1%. The PLS229 genome was annotated and has 3,259 open reading frames and 50 RNA genes. PMID:24652975

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

    OpenAIRE

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

    2015-01-01

    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 bar...

  14. Analysis of the drying kinetics of S. Bartolomeu pears for different drying systems.

    OpenAIRE

    Guiné, Raquel

    2010-01-01

    Pears of the variety S. Bartolomeu (Pyrus communis L.) are used in Portugal to produce a traditional food product, which is obtained by open-air sun drying. The traditional processing has many disadvantages, namely the efficiency and the sanitary quality of the final product. For these reasons, attempts have been made to study alternative production methodologies, including the use of a solar stove, a solar drier and a drying tunnel. In the present work pears of the variety S. Bartolomeu,...

  15. Processed Kaolin as an alternative insecticide against the European pear sucker, Cacopsylla pyri (L.)

    OpenAIRE

    Daniel, Claudia; Pfammatter, W; Kehrli, P; Wyss, Eric

    2005-01-01

    Application of processed kaolin particle film repels insects without lethal effects; hence side effects on beneficial arthropods are low. Processed kaolin may be an alternative to broad-spectrum insecticides used against European pear sucker, Cacopsylla pyri (L.), in organic and conventional pear production. Trials showed that the processed kaolin showed a very high efficacy, and the population of C. pyri was kept under a damaging level over the whole season. In conclusion, kaolin shows p...

  16. Effects and action mechanisms of Korean pear (Pyrus pyrifolia cv. Shingo) on alcohol detoxification.

    Science.gov (United States)

    Lee, Ho-Sun; Isse, Toyoshi; Kawamoto, Toshihiro; Woo, Hyun-Su; Kim, An Keun; Park, Jong Y; Yang, Mihi

    2012-11-01

    Korean pear (Pyrus pyrifolia cv. Shingo) has been used as a traditional medicine for alleviating alcohol hangover. However, scientific evidence for its effectiveness or mechanism is not clearly established. To investigate its mechanism of alcohol detoxification, both in vitro and in vivo studies were performed with an aldehyde dehydrogenase 2 (ALDH2) alternated animal model. The pear extract (10 mL/kg bw) was administered to Aldh2 normal (C57BL/6) and deficient (Aldh2 -/-) male mice. After 30 min, ethanol (1 g or 2 g/kg bw) was administered to the mice via gavage. Levels of alcohol and acetaldehyde in blood were quantified by GC/MS. First, it was observed that the pears stimulated both alcohol dehydrogenase (ADH) and ALDH activities by 2∼3-  and 1.3-fold in in vitro studies, respectively. Second, mouse PK data (AUC(∞) and C(max) ) showed that the pear extract decreased the alcohol level in blood regardless of ALDH2 genotype. Third, the pear increased the acetaldehyde level in blood in Aldh2 deficient mice but not in Aldh2 normal mice. Therefore, the consistent in vitro and in vivo data suggest that Korean pears stimulate the two key alcohol-metabolizing enzymes. These stimulations could be the main mechanism of the Korean pear for alcohol detoxification. Finally, the results suggest that polymorphisms of human ALDH2 could bring out individual variations in the effects of Korean pear on alcohol detoxification. PMID:22451246

  17. Life cycle assessment of fossil energy use and greenhouse gas emissions in Chinese pear production

    OpenAIRE

    Liu, Y.; Langer, V; Høgh-Jensen, H.; Egelyng, H.

    2010-01-01

    An environmental life cycle assessment (LCA) was performed in China to investigate environmental consequences of the life cycle of pears in terms of fossil energy use and greenhouse gas emissions. The assessment identified activities that contributed significantly to pears’ environmental impacts from the cradle to the point of sale. Cultivation was identified as having the greatest greenhouse gas emissions in pear production chains, followed by the process of storage and transportation. The ...

  18. 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

  19. Extending the Shelf-life of Pear Fruits by Using Gamma Irradiation

    International Nuclear Information System (INIS)

    The effects of gamma irradiation on lesion diameters of grey mold disease of pear and some physico-chemical parameters of pear fruits were studied in this investigation as a pre storage treatment for extend shelf life of pear fruits. Pear fruits (Pyrus Communis, L.) were exposed to different gamma irradiation doses 0, 0.5, 1, 1.5, 2, 2.5 and 3 KGy and stored for 10, 20 and 30 days in 0 degree C and 85-90% RH. The increment of irradiant dose caused decrement of lesion diameters of grey mold disease of pear. The narrowest diameters were recorded with 3.0 KGy irradiation dose. The low dose of gamma irradiation 0.5 KGy gave a relatively high value of firmness although in higher doses firmness of pear fruits decreased. The highest loss weight was found in unirradiated fruits while the loss of weight in all irradiated fruits was still lower than those of unirradiated ones and low radiation doses decreased the loss of weight. Total soluble solid increased with increasing in storage periods with respect to gamma irradiation effect there was fluctuation in total soluble solid values. There was a decreasing in acidity during storage of irradiated and un-irradiated fruits. Free amino acid and soluble protein show a slight increasing at 0.5 KGy such increasing was followed by a gradual decrease in higher gamma irradiation doses

  20. Relative effect of temperature and pH on diel cycling of dissolved trace elements in prickly pear creek, Montana

    Science.gov (United States)

    Jones, C.A.; Nimick, D.A.; McCleskey, R.B.

    2004-01-01

    Diel (24 hr) cycles in dissolved metal and As concentrations have been documented in many northern Rocky Mountain streams in the U.S.A. The cause(s) of the cycles are unknown, although temperature- and pH-dependent sorption reactions have been cited as likely causes. A light/dark experiment was conducted to isolate temperature and pH as variables affecting diel metal cycles in Prickly Pear Creek, Montana. Light and dark chambers containing sediment and a strand of macrophyte were placed in the stream to simulate instream temperature oscillations. Photosynthesis-induced pH changes were allowed to proceed in the light chambers while photosynthesis was prevented in the dark chambers. Water samples were collected periodically for 22 hr in late July 2001 from all chambers and the stream. In the stream, dissolved Zn concentrations increased by 300% from late afternoon to early morning, while dissolved As concentrations exhibited the opposite pattern, increasing 33% between early morning and late afternoon. Zn and As concentrations in the light chambers showed similar, though less pronounced, diel variations. Conversely, Zn and As concentrations in the dark chambers had no obvious diel variation, indicating that light, or light-induced reactions, caused the variation. Temperature oscillations were nearly identical between light and dark chambers, strongly suggesting that temperature was not controlling the diel variations. As expected, pH was negatively correlated (P pH changes were determined to be the major cause of the diel dissolved Zn and As cycles in Prickly Pear Creek. Further research is necessary in other streams to verify that this finding is consistent among streams having large differences in trace-element concentrations and mineralogy of channel substrate. ?? 2004 Kluwer Academic Publishers.

  1. 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)

  2. 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

  3. 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

  4. 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

  5. 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.

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

    Directory of Open Access Journals (Sweden)

    Lina Marcela Villa

    2013-11-01

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

  7. Erwinia pyrifoliae sp. nov., a novel pathogen that affects Asian pear trees (Pyrus pyrifolia Nakai)

    Science.gov (United States)

    Kim, W S; Gardan, L; Rhim, S L; Geider, K

    1999-04-01

    A novel pathogen from Asian pears (Pyrus pyrifolia Nakai) was analysed by sequencing the 16S rDNA and the adjacent intergenic region, and the data were compared to related Enterobacteriaceae. The 16S rDNA of the Asian pear pathogen was almost identical with the sequence of Erwinia amylovora, in contrast to the 16S-23S rRNA intergenic transcribed spacer region of both species. A dendrogram was deduced from determined sequences of the spacer regions including those of several related species such as Erwinia amylovora, Enterobacter pyrinus, Pantoea stewartii subsp. stewartii and Escherichia coli. Dendrograms derived from 121 biochemical characteristics including Biotype 100 data placed the Asian pear pathogen close to Erwinia amylovora and more distantly to other members of the species Erwinia and to the species Pantoea and Enterobacter. Another DNA relatedness study was performed by DNA hybridizations and estimation of delta Tm values. The Asian pear strains constituted a tight DNA hybridization group (89-100%) and were barely related to strains of Erwinia amylovora (40-50%) with a delta Tm in the range of 5.2-6.8. The G + C content of DNA from the novel pathogen is 52 mol%. Therefore, it is proposed that strains isolated from Asian pears constitute a new species and the name Erwinia pyrifoliae is suggested; the type strain is strain Ep 16/96T (= CFBP 4172T = DSM 12163T). PMID:10319516

  8. 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. PMID:23545106

  9. 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.

  10. 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.

  11. Complete Genome Sequence of Japanese Erwinia Strain Ejp617, a Bacterial Shoot Blight Pathogen of Pear

    OpenAIRE

    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

    2010-01-01

    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.

  12. Identification of a Simple Sequence Repeat molecular-marker set for large-scale analyses of pear germplasm

    Directory of Open Access Journals (Sweden)

    Gabriel Dequigiovanni

    2012-01-01

    Full Text Available Simple Sequence Repeats (SSR are molecular markers suitable to assess the genetic variation of germplasm resources; however, large-scale SSR use requires protocol optimization. The present work aimed to identify SSR markers, developed for pear and other fruit species that are effective in characterizing pear germplasm collections and in demonstrating their use in providing support for genetic breeding programs. From a total of 62 SSR markers investigated, 23 yielding reproducible and polymorphic patterns were used to genotype a sample of 42 pear accessions of the Brazilian Pear Germplasm Bank (PGB. When compared to these 23 SSR markers, a subset of eleven markers, selected based on He, PIC and PId, was used to distinguish individual accessions and perform cluster analysis with similar efficacy. Genetic diversity analysis clustered the European, Japanese and Chinese accessions in distinct groups. This markers subset constitutes a valuable tool for several applications related to pear genetic resources management and breeding.

  13. 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...

  14. 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...

  15. 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.

  16. A 'compare and contrast' exercise: wrapping versus personalised external aortic root support (PEARS).

    Science.gov (United States)

    Treasure, Tom

    2016-01-01

    Wrapping of the aorta and personalised external aortic root support (PEARS) both have the purpose of preventing further expansion of the ascending aorta in order to reduce the risk of aortic dissection and to spare the patient the disastrous consequences of aortic rupture. For the first time, Plonek and colleagues have reported systematically the CT appearances of a series of cases of wrapping. They illustrate the important finding that there are residual spaces between the aorta and the wrap. PEARS by contrast is intimately in contact with the aorta due to its personalised design and is fully incorporated due it construction from a porous mesh. A limitation of PEARS is that it is, of its nature, a planned and elective operation while wrapping can be undertaken during an emergency operation and can be used without prior planning as an intraoperative decision. PMID:27406033

  17. 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.

  18. 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.

  19. 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.

  20. 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.

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

    OpenAIRE

    Mateus da Silveira Pasa; José Carlos Fachinello; Horacy Fagundes da Rosa Júnior; Émerson De Franceschi; Juliano Dutra Schmitz; André Luiz Kulkamp de Souza

    2015-01-01

    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 frui...

  2. 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.

  3. Optical chirped beam amplification and propagation

    Science.gov (United States)

    Barty, Christopher P.

    2004-10-12

    A short pulse laser system uses dispersive optics in a chirped-beam amplification architecture to produce high peak power pulses and high peak intensities without the potential for intensity dependent damage to downstream optical components after amplification.

  4. 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.

  5. Double regenerative amplification of picosecond pulses

    Science.gov (United States)

    Bai, Zhen-ao; Chen, Li-yuan; Bai, Zhen-xu; Chen, Meng; Li, Gang

    2012-04-01

    An double Nd:YAG regenerative amplification picosecond pulse laser is demonstrated under the semiconductor saturable absorption mirror(SESAM) mode-locking technology and regenerative amplification technology, using BBO crystal as PC electro-optic crystal. The laser obtained is 20.71ps pulse width at 10 KHz repetition rate, and the energy power is up to 4W which is much larger than the system without pre-amplification. This result will lay a foundation for the following amplification.

  6. 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.

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

    DEFF Research Database (Denmark)

    Wu, Jun; Wang, Zhiwen; Shi, Zebin;

    2013-01-01

    to the ancestor of Rosaceae has reconstructed nine ancestral chromosomes. Pear and apple diverged from each other ~5.4-21.5 million years ago, and a recent whole-genome duplication (WGD) event must have occurred 30-45 MYA prior to their divergence, but following divergence from strawberry. When compared...

  8. 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:...

  9. 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. PMID:22899620

  10. 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.

  11. Enriched antioxidant activity of pear juice by supplementation with oregano and wild thyme extracts

    Directory of Open Access Journals (Sweden)

    Tudor Lucian MIRON

    2012-12-01

    Full Text Available In the last years, it has been noticed an increased interest in natural antioxidants and the use of these compounds in the field of new polyphenol-rich drinks. Antioxidants are compounds responsible for free radical scavenging in the body. They protect the organism from oxidative modification of cells and tissues. The aim of this study was to improve the antioxidant capacity of a pear juice, as well as its compounds stability during 12 days of storage. The stabilization of phenols and their delivery in pear juice was possible thanks to microemulsions obtained by mixing different ratios of oil, surfactant and cosurfactant. The characterization ofmicroemulsions was performed using pseudoternary phase diagrams and which have highlighted reports of mixing A / W / S / CoS corresponding states of microemulsion. The obtained microemulsions have stability and provides good stabilization of linoleic acid and walnuit oil, up to 70% w/w water. Through conductometric analysis we have highlighted the transition states of micro W/O, O/W bicontinuous structures. The pear juice with plant extracts had higher antioxidant values than the pear juice with synthetic antioxidants. Thus the juice with oregano improved the antioxidant capacity approximately 1.45 times compared to the juice with BHA and approximately 1.67 times compared to the juice with BHT. These results are promising in order to replace the synthetic antioxidants with natural antioxidants.

  12. Effect of the yeast Rhodosporidium paludigenum on postharvest decay and patulin accumulation in apples and pears.

    Science.gov (United States)

    Zhu, Ruiyu; Yu, Ting; Guo, Shuanghuan; Hu, Hao; Zheng, Xiaodong; Karlovsky, Petr

    2015-01-01

    The effect of a strain of marine yeast Rhodosporidium paludigenum on postharvest blue mold and patulin accumulation in apples and pears stored at 23°C was evaluated. The occurrence and severity of apple and pear decay caused by Penicillium expansum were significantly inhibited by R. paludigenum. However, the application of the yeast at a high concentration (10(8) cells per ml) enhanced patulin accumulation after 7 days of storage; the amount of patulin increased 24.2 times and 12.6 times compared to the controls in infected apples and pears, respectively. However, R. paludigenum reduced the patulin concentration in the growth medium by both biological degradation and physical adsorption. Optimal in vitro patulin reduction was observed at 30°C and at pH 6.0. R. paludigenum incubated at 28°C was tolerant to patulin at concentrations up to 100 mg/liter. In conclusion, R. paludigenum was able to control postharvest decay in apples and pears and to remove patulin in vitro effectively. However, because the yeast induced patulin accumulation in fruit, the assessment of mycotoxin content after biological treatments in postharvest decay control is important. R. paludigenum may also be a promising source of gene(s) and enzyme(s) for patulin degradation and may be a tool to decrease patulin contamination in commercial fruit-derived products.

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

    Directory of Open Access Journals (Sweden)

    Mostafa GHASEMI

    2011-03-01

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

  14. 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.

  15. Dissipation of teflubenzuron and triflumuron residues in field-sprayed and cold-stored pears.

    Science.gov (United States)

    Aplada-Sarlis, P G; Miliadis, G E; Tsiropoulos, N G

    1999-07-01

    Dissipation of residues of benzoylurea insecticides teflubenzuron (TFB) and triflumuron (TFM) under field conditions was evaluated on a pear orchard in Greece. Residues were determined by UV-HPLC analysis, with a detection limit of 0.030 mg/kg for both pesticides. TFB residues in pears were found to persist for 2 weeks and decline thereafter with 48% of the initial deposit remaining 42 days after the last application. TFM residues were found to decline following first-order kinetics and with a half-life of 39(+/-7) days. Residues of both pesticides found in pears collected at harvest maturity were lower than the maximum residue limits (MRLs) set by individual countries. Dissipation of TFB and TFM in cold-stored pears was also evaluated. TFB residues were very persistent for the whole storage period, whereas TFM residues did not dissipate for 6 weeks and then showed a constant decline; 7% of the initial concentration remained at the end of the storage period of 29 weeks. PMID:10552588

  16. Regulating Ripening of 'Bartlett' Pears Using Preharvest Plus Postharvest Aminoethoxyvinylglycine (AVG)

    Science.gov (United States)

    Variation in ripening uniformity of Bartlett (Pyrus Communis) pears in cold storage may be a problem in some years because of factors such as lack of chilling hours during winter, protracted anthesis and insufficient labor at harvest. On large orchards, such variation makes harvest timing very diffi...

  17. 软儿梨的研究进展%Progress Research of Ruan’er Pear

    Institute of Scientific and Technical Information of China (English)

    田晓菊

    2015-01-01

    软儿梨营养丰富,食用时将其冷冻、解冻后吸食果汁,而且具有很多保健功能,但是长期以来缺乏对其生理特性的研究,使这种资源不受重视而越来越少。本文就软儿梨目前国内的研究现状进行阐述,以期为软儿梨的开发利用提供依据。%There is much nutrition in Ruan’er pear which is a kind of soft pear with much juice after freezing and defrosting and also has much health function. However, because research in its physiological property over time is limited, the resource of Ruan’er pear, with less attention, is on the decrease. The current research status of Ruan’er pear interiorly has been elaborated in the paper in order to provide a basis for its development and utilization.

  18. Comprehensive human genome amplification using multiple displacement amplification

    OpenAIRE

    Dean, Frank B.; Hosono, Seiyu; Fang, Linhua; Wu, Xiaohong; Faruqi, A. Fawad; Bray-Ward, Patricia; Zhenyu SUN; Zong, Qiuling; Du, Yuefen; Du, Jing; Driscoll, Mark; Song, Wanmin; Kingsmore, Stephen F.; Egholm, Michael; Lasken, Roger S.

    2002-01-01

    Fundamental to most genetic analysis is availability of genomic DNA of adequate quality and quantity. Because DNA yield from human samples is frequently limiting, much effort has been invested in developing methods for whole genome amplification (WGA) by random or degenerate oligonucleotide-primed PCR. However, existing WGA methods like degenerate oligonucleotide-primed PCR suffer from incomplete coverage and inadequate average DNA size. We describe a method, termed multi...

  19. 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

  20. Rapid Amplification of Plasmid and Phage DNA Using Phi29 DNA Polymerase and Multiply-Primed Rolling Circle Amplification

    OpenAIRE

    Dean, Frank B.; Nelson, John R.; Giesler, Theresa L.; Lasken, Roger S.

    2001-01-01

    We describe a simple method of using rolling circle amplification to amplify vector DNA such as M13 or plasmid DNA from single colonies or plaques. Using random primers and φ29 DNA polymerase, circular DNA templates can be amplified 10,000-fold in a few hours. This procedure removes the need for lengthy growth periods and traditional DNA isolation methods. Reaction products can be used directly for DNA sequencing after phosphatase treatment to inactivate unincorporated nucleotides. Amplified ...

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

    International Nuclear Information System (INIS)

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

  2. 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.

  3. 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.

  4. 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.

  5. Hybrid chirped pulse amplification system

    Science.gov (United States)

    Barty, Christopher P.; Jovanovic, Igor

    2005-03-29

    A hybrid chirped pulse amplification system wherein a short-pulse oscillator generates an oscillator pulse. The oscillator pulse is stretched to produce a stretched oscillator seed pulse. A pump laser generates a pump laser pulse. The stretched oscillator seed pulse and the pump laser pulse are directed into an optical parametric amplifier producing an optical parametric amplifier output amplified signal pulse and an optical parametric amplifier output unconverted pump pulse. The optical parametric amplifier output amplified signal pulse and the optical parametric amplifier output laser pulse are directed into a laser amplifier producing a laser amplifier output pulse. The laser amplifier output pulse is compressed to produce a recompressed hybrid chirped pulse amplification pulse.

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

    OpenAIRE

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

    2009-01-01

    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. Res...

  7. 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.

  8. Transient amplification limits noise suppression in biochemical networks

    Science.gov (United States)

    Dixon, John; Lindemann, Anika; McCoy, Jonathan H.

    2016-01-01

    Cell physiology is orchestrated, on a molecular level, through complex networks of biochemical reactions. The propagation of random fluctuations through these networks can significantly impact cell behavior, raising challenging questions about how network design shapes the cell's ability to suppress or exploit these fluctuations. Here, drawing on insights from statistical physics, fluid dynamics, and systems biology, we explore how transient amplification phenomena arising from network connectivity naturally limit a biochemical system's ability to suppress small fluctuations around steady-state behaviors. We find that even a simple system consisting of two variables linked by a single interaction is capable of amplifying small fluctuations orders of magnitude beyond the levels predicted by linear stability theory. We also find that adding additional interactions can promote further amplification, even when these interactions implement classic design strategies known to suppress fluctuations. These results establish that transient amplification is an essential factor determining baseline noise levels in stable intracellular networks. Significantly, our analysis is not bound to specific systems or interaction mechanisms: we find that noise amplification is an emergent phenomenon found near steady states in any network containing sufficiently strong interactions, regardless of its form or function.

  9. 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.

  10. 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

    2014-08-01

    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.

  11. 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.

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

    Science.gov (United States)

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

    2015-01-01

    Changes in chemical constituent contents and DPPH radical-scavenging activity in fruits of pear (Pyrus pyrifolia) cultivars during the development were investigated. The fruits of seven cultivars (cv. Niitaka, Chuhwangbae, Wonhwang, Hwangkeumbae, Hwasan, Manpungbae, and Imamuraaki) were collected at 15-day intervals after day 20 of florescence. Vitamins (ascorbic acid and α-tocopherol), arbutin, chlorogenic acid, malaxinic acid, total caffeic acid, total flavonoids, and total phenolics were the highest in immature pear fruit on day 20 after florescence among samples at different growth stages. All of these compounds decreased gradually in the fruit during the development. Immature pear fruit on day 35 or 50 after florescence exhibited higher free radical-scavenging activity than that at other times, although activities were slightly different among cultivars. The chemical constituent contents and free radical-scavenging activity were largely different among immature fruits of the pear cultivars, but small differences were observed when they matured. PMID:25348501

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

    OpenAIRE

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

    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...

  14. Effcet of ripening stage on the solar drying kinetics and properties of S. Bartolomeu pears (Pirus Communis L.).

    OpenAIRE

    Guiné, Raquel; Lopes, Pedro; Barroca, Maria João; Ferreira, Dulcineia

    2009-01-01

    Pears of the variety S. Bartolomeu (Pyrus communis L.) have been used over the years in Portugal to produce a traditional dried pear known as “pera passa”. The processing comprehends a solar drying performed at open air, with obvious disadvantages, either concerning the drying efficiency or the sanitary quality of the final product, taking into account that the products are exposed to multiple contamination agents. For these reasons, attempts have been made to study alternative production met...

  15. Heralded amplification of photonic qubits.

    Science.gov (United States)

    Bruno, Natalia; Pini, Vittorio; Martin, Anthony; Verma, Varun B; Nam, Sae Woo; Mirin, Richard; Lita, Adriana; Marsili, Francesco; Korzh, Boris; Bussières, Félix; Sangouard, Nicolas; Zbinden, Hugo; Gisin, Nicolas; Thew, Rob

    2016-01-11

    We demonstrate postselection free heralded qubit amplification for Time-Bin qubits and single photon states in an all-fibre, telecom-wavelength, scheme that highlights the simplicity, stability and potential for fully integrated photonic solutions. Exploiting high-efficiency superconducting detectors, the gain, fidelity and the performance of the amplifier are studied as a function of loss. We also demonstrate the first heralded single photon amplifier with independent sources. This provides a significant advance towards demonstrating device-independent quantum key distribution as well as fundamental tests of quantum mechanics over extended distances. PMID:26832244

  16. Resonant primordial gravitational waves amplification

    Directory of Open Access Journals (Sweden)

    Chunshan Lin

    2016-01-01

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

  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 of target-specific, ligation-dependent circular probe.

    Science.gov (United States)

    Zhang, D Y; Brandwein, M; Hsuih, T C; Li, H

    1998-05-12

    We describe a novel polymerase chain reaction (PCR)-based gene amplification method utilizing a circularizable oligodeoxyribonucleotide probe (C-probe). The C-probe contains two target complementary regions located at each terminus and an interposed generic PCR primer binding region. The hybridization of C-probe to a target brings two termini in direct apposition as the complementary regions of C-probe wind around the target to form a double helix. Subsequent ligation of the two termini results in a covalently linked C-probe that becomes 'locked on to' the target. The circular nature of the C-probe allows for the generation of a multimeric single-stranded DNA (ssDNA) via extension of the antisense primer by Taq DNA polymerase along the C-probe and displacement of downstream strand, analogous to 'rolling circle' replication of bacteriophage in vivo. This multimeric ssDNA then serves as a template for multiple sense primers to hybridize, extend, and displace downstream DNA, generating a large ramified (branching) DNA complex. Subsequent thermocycling denatures the dsDNA and initiates the next round of primer extension and ramification. This model results in significantly improved amplification kinetics (super-exponential) as compared to conventional PCR. Our results show that the C-probe was 1000 times more sensitive than the corresponding linear hemiprobes for detecting Epstein-Barr virus early RNA. The C-probe not only increases the power of amplification but also offers a means for decontaminating carryover amplicons. As the ligated C-probes possess no free termini, they are resistant to exonuclease digestion, whereas contaminated linear amplicons are susceptible to digestion. Treatment of the ligation reaction mixture with exonuclease prior to amplification eliminated the amplicon contaminant, which could also have been co-amplified with the same PCR primers; only the ligated C-probes were amplified. The combined advantages of the C-probe and thermocycling have a

  1. 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.

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

    Directory of Open Access Journals (Sweden)

    Mateus da Silveira Pasa

    2015-02-01

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

  3. 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.

  4. 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.

  5. Whole Transcriptome Amplification for Gene Expression Profiling and Development of Molecular Archives

    Directory of Open Access Journals (Sweden)

    Scott A. Tomlins

    2006-02-01

    Full Text Available Expression profiling of clinically obtainable tumor specimens has been hindered by the need for microgram quantities of RNA. In vitro transcription (IVT-based amplifications are most commonly used to amplify small quantities of RNA for microarray analysis. However, significant drawbacks exist with IVT-based amplification, and the need for alternative amplification methods remains. Herein, we validate whole transcriptome amplification (WTA, an exponential amplification technique that produces cDNA libraries and amplified target in 3 to 4 hours from nanogram quantities of total RNA using a combination of cDNA microarrays and quantitative polymerase chain reaction (PCR. We demonstrate that WTA material can serve as a “molecular archive” because a WTA cDNA library can be faithfully amplified through multiple rounds of PCR amplification, allowing it to serve as a bankable and distributable resource. To demonstrate applicability, WTA was combined with laser capture microdissection to profile frozen prostate tissues. Unlike most IVT-based and exponential amplification techniques, WTA does not depend on the presence of a poly-A tail. Thus, we demonstrate that WTA is compatible with artificially degraded RNA and RNA isolated from formalin-fixed paraffin-embedded tissues. Taken together, WTA represents a versatile approach to profile and archive cDNA from minute tumor samples and is compatible with partially degraded RNA.

  6. Study on Pear Diseases Query System Based on Ontology and SWRL

    OpenAIRE

    Sun, Qian; Liang, Yong

    2013-01-01

    International audience This paper studied the construction of Pear Diseases Domain Ontology (PDDO), and the realization of query system based on PDDO and SWRL. First, an approach to build PDDO based on SWRL was proposed, which consists of confirming core concepts, adding the properties of concepts and the relationships between concepts, adding the instances of concepts, representing domain ontology, adding SWRL rules and reasoning. Then the query system model and implementation algorithm w...

  7. 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; Mariane LUTZ

    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...

  8. Variation of textural properties in pears dried by different solar methodologies.

    OpenAIRE

    Abílio, Sandra; Guiné, Raquel; Sousa, Isabel

    2011-01-01

    In Portugal, the variety of S. Bartholomew (Pyrus communis L.) pears are subject to an artisan drying process consisting of direct open-air sun exposure, leading to a traditional product with unique texture characteristics, called “Pêra Passa de Viseu”. However, the drying process does not provide the current standards of safety and, therefore, recent investigations have emerged with alternatives to the traditional drying process. The changes that occur in t...

  9. Comparative study of pear drying using solar stove and tunnel drying

    OpenAIRE

    Guiné, Raquel; Barroca, Maria João; Lopes, Paulo; Silva, Vitor; Lima, Maria João; Ferreira, Dulcineia

    2010-01-01

    Sun-dried pears of Portuguese S. Bartolomeu variety are relatively small fruits with peculiar organolephtic characteristics. The open sun drying has some disadvantages related with drying efficiency and food safety. However the drying with an economic and friendly energy is an important characteristic of the drying methods and therefore several attempts have been made to develop processes using solar drying. The present work studies two ...

  10. Osmia cornuta (Hymenoptera, Megachilidae) as a pollinator of pear (Pyrus communis): fruit- and seed-set

    OpenAIRE

    Maccagnani, Bettina; Ladurner, Edith; SANTI, FABRIZIO; Burgio, Giovanni

    2003-01-01

    To investigate the possibility of pollinating 'Abate Fetel' and 'Max Red Bartlett' pears with O. cornuta, different pollination treatments were compared in 1998 and 1999: (1) trees caged with O. cornuta bees; (2) open pollination - uncaged trees located at various distances from O. cornuta nesting shelters; (3) trees caged without bees. O. cornuta was the most abundant pollinator species in the orchard (97.6% of the observed insects). Caged trees pollinated by O. cornuta set significantly mor...

  11. Isolation and Identification of the Antioxidant DDMP from Heated Pear (Pyrus pyrifolia Nakai)

    OpenAIRE

    Hwang, In Guk; Kim, Hyun Young; Woo, Koan Sik; Lee, Sang Hoon; Lee, Junsoo; Jeong, Heon Sang

    2013-01-01

    We evaluated antioxidant activities of heated pear juice (HPJ) exposed to 120, 130, and 140°C for 2 hr. HPJ was partitioned using n-hexane, chloroform, ethyl acetate, n-butanol, and water. The ethyl acetate fraction treated at 130°C for 2 hr showed strong antioxidant activity; thus, this extract was isolated and purified using silica gel column chromatography and preparative high performance liquid chromatography. The structure of the purified compound was determined using ultraviolet and mas...

  12. Graft Incompatibility Influence on Assimilating Pigments and Soluble Sugars Amount of some Pear (Pyrus sativa) Cultivars

    OpenAIRE

    Gheorghii CIOBOTARI; Maria BRINZA; Aliona MORARIU; Gica GRADINARIU

    2010-01-01

    Graft incompatibility in fruit trees is one of the greatest obstacles in rootstocks and cultivars breeding. The mechanism in which incompatibility is expressed is not yet fully understood and several hypotheses have been advanced in an attempt to explain it. In many cases (pear on quince grafts, apricot on Prunus grafts), incompatibility is manifested by the breaking of the trees at the point of the union particularly when they have been growing for some years. Many reports focus on this prob...

  13. The Variability of Fruit Characteristics of Traditional Pear Karamanka in Different Ecological Conditions

    OpenAIRE

    Selamovska, A; Miskoska – Milevska, E; Dimovska, Violeta; Najdenovska, O

    2014-01-01

    In this paper, we present the results of the phenological characteristics (flowering and ripening time), fruit characteristics (fruit mass, fruit length, fruit width, fruit hardness, length of fruit stalk and number of seeds in fruit) and chemical characteristics of fruits (soluble dry matter, total sugar and total acids) of a traditional pear variety ‘Karamanka’. For this study, a survey was undertaken in different ecological regions of Macedonia i.e. Skopje, Kumanovo, Kratovo, Kriva Pala...

  14. Efficacy of different insecticides and a repellent against the European pear sucker (Cacopsylla pyri)

    OpenAIRE

    Daniel, Claudia; Wyss, Eric

    2004-01-01

    The efficacy of different insecticides (neem, pyrethrin, spinosad, and rotenone) and a repellent (kaolin) applied with different strategies (single or repeated applications) against the over-wintering pear suckers (Cacopsylla pyri) and nymphs of the first generation was tested in a field trial in Switzerland in spring 2003. Rotenone, the only admitted product in Swiss organic agriculture, showed good effects. But, since Rotenone is toxic for non-target insects we looked for an ...

  15. Türgi võimupartei vihastab pearätivastaseid / Liisi Poll

    Index Scriptorium Estoniae

    Poll, Liisi, 1980-

    2007-01-01

    Ilmunud ka: Postimees : na russkom jazõke, 4. okt. 2007, lk. 7. Türgi naisõiguslased ründavad peaminister Recep Tayyip Erdogani islamipartei põhiseaduse eelnõu, mis ei kaitse piisavalt naiste õigusi; lisaks on peaminister pahameelt tekitanud pearätiku kandmist kaitstes. Vt samas: Väärarusaamad kimbutavad Euroopa Liidu ja Türgi suhteid

  16. Risk Perception and Social Amplification

    International Nuclear Information System (INIS)

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

  17. 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.

  18. Genetic structure and diversity of the wild Ussurian pear in East Asia.

    Science.gov (United States)

    Katayama, Hironori; Amo, Hitomi; Wuyun, Tana; Uematsu, Chiyomi; Iketani, Hiroyuki

    2016-01-01

    The Ussurian pear is the most important cultivated pear in the northern part of China. Cultivated Ussurian pears are considered to have derived from Pyrus ussuriensis Maxim. which is native to the northeast of China. In Japan, two varieties of P. ussuriensis, P. ussuriensis var. aromatica and var. hondoensis are native to the northern area and the central area of the main island respectively. In order to reveal the origin of Pyrus ussuriensis var. aromatica distributed in the northern area of main island of Japan, more than 40 explorations have been performed in Japan and in China, and more than 30 natural habitats were recognized. These natural habitats are at risk of extinction because of human development and forest degradation caused by climate change. Population structure and genetic diversity of P. ussuriensis in China and P. ussuriensis var. aromatica in Japan have been investigated using both morphological and molecular markers in order to define appropriate conservation units, and to provide a good focus for conservation management. Distant evolutionary relationships between P. ussuriensis Maxim. in China and P. ussuriensis var. aromatica in Japan inferred from population genetic structure and phylogenetic analysis are also discussed. PMID:27069394

  19. Spectroscopic Study of the HST/ACS PEARS Emission-Line Galaxies

    CERN Document Server

    Xia, Lifang; Rhoads, James; Pirzkal, Norbert; Meurer, Gerhardt; Straughn, Amber; Floyd, David; Zheng, Zhenya

    2010-01-01

    We present spectroscopy of 76 emission-line galaxies (ELGs) in CDF-S taken with the LDSS3 spectrograph on Magellan Telescope. These galaxies are selected to have emission lines with ACS grism data in the Hubble Space Telescope Probing Evolution and Reionization Spectroscopically (PEARS) grism Survey. While the ACS grism spectra cover the wavelength range 6000-9700 {\\AA} and most PEARS grism redshifts are based on a single emission line + photometric redshifts from broad-band colors; the Magellan spectra cover a wavelength range of 4000 {\\AA} to 9000 {\\AA}, and provide a check on redshifts derived from PEARS data. We find an accuracy of {\\sigma}z = 0.006 for the grism redshifts with only one catastrophic outlier. For 14 galaxies at z 10$^{43} ergs s$^{-1}, and power-law continuum spectra. Three objects are identified as starburst galaxies from the full-band X-ray luminosity L$_{FB} ~ 10$^{41}$ ergs s$^{-1}.

  20. 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.

  1. 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)

  2. 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.

  3. 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.

  4. Tsunami Amplification due to Focusing

    Science.gov (United States)

    Moore, C. W.; Kanoglu, U.; Titov, V. V.; Aydin, B.; Spillane, M. C.; Synolakis, C. E.

    2012-12-01

    Tsunami runup measurements over the periphery of the Pacific Ocean after the devastating Great Japan tsunami of 11 March 2011 showed considerable variation in far-field and near-field impact. This variation of tsunami impact have been attributed to either directivity of the source or by local topographic effects. Directivity arguments alone, however, cannot explain the complexity of the radiated patterns in oceans with trenches and seamounts. Berry (2007, Proc. R. Soc. Lond. A 463, 3055-3071) discovered how such underwater features may concentrate tsunamis into cusped caustics and thus cause large local amplifications at specific focal points. Here, we examine focusing and local amplification, not by considering the effects of underwater diffractive lenses, but by considering the details of the dipole nature of the initial profile, and propose that certain regions of coastline are more at-risk, not simply because of directivity but because typical tsunami deformations create focal regions where abnormal tsunami wave height can be registered (Marchuk and Titov, 1989, Proc. IUGG/IOC International Tsunami Symposium, Novosibirsk, USSR). In this work, we present a new general analytical solution of the linear shallow-water wave equation for the propagation of a finite-crest-length source over a constant depth without any restriction on the initial profile. Unlike the analytical solution of Carrier and Yeh (2005, Comp. Mod. Eng. & Sci. 10(2), 113-121) which was restricted to initial conditions with Gaussian profiles and involved approximation, our solution is not only exact, but also general and allows the use of realistic initial waveform such as N-waves as defined by Tadepalli and Synolakis (1994, Proc. R. Soc. Lond. A 445, 99-112). We then verify our analytical solution for several typical wave profiles, both with the NOAA tsunami forecast model MOST (Titov and Synolakis, 1998, J. Waterw. Port Coast. Ocean Eng. 124(4), 157-171) which is validated and verified through

  5. 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

  6. 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.

  7. Mechanisms of Metal-Induced Centrosome Amplification

    OpenAIRE

    Holmes, Amie L.; Wise, John Pierce

    2010-01-01

    Exposure to toxic and carcinogenic metals is widespread; however, their mechanisms of action remain largely unknown. One potential mechanism for metal-induced carcinogenicity and toxicity is centrosome amplification. Here, we review the mechanisms for metal-induced centrosome amplification, including arsenic, chromium, mercury and nano-titanium dioxide.

  8. Risk Perception and Social Amplification

    Energy Technology Data Exchange (ETDEWEB)

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

    2001-07-01

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

  9. 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.

  10. 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.

  11. 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. PMID:23272258

  12. Effect of low and superatmosferic O2 modified atmosphere on the quality of fresh-cut pears

    Directory of Open Access Journals (Sweden)

    Slavica Grujic

    2010-08-01

    Full Text Available Long-term performance of structures has become vital to the economies of all nations. Concrete has been the major instrument for providing stable and reliable The effect of low O2 and superatmosferic O2 atmospheres in combination with a dipping into N-acetyl-L-cysteine (NAC and reduced glutathione (GSH on fresh-like quality “Flor de Invierno” pears was studied. Changes in headspace gas composition, colour, firmness, acidity and soluble solids were examined. The fresh-cut pears were dipped into 0.75% (w/v NAC and 0.75% (w/v GSH aqueous solution and then packaged into polypropylene trays under low O2, superatmosferic O2 and traditional passive atmosphere (PA conditions. Trays were stored at 4±0.5 0C and analyzed periodically during 28 days. Superatmosferic O2 atmosphere caused a stress in pears cells tissue leading to many contents of ethylene, acetaldehyde and ethanol and limiting shelf-life of pears up to 21 days. Because of complete inhibition of ethylene production, maintenance of initial colour, texture and acidity, a low O2 atmosphere was the most effective for fresh-cut pears quality preserving during 28 days of refrigerated storage.

  13. 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.

  14. 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.

  15. 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.

  16. Effect of gamma-irradiation and refrigerated storage on the improvement of quality and shelf life of pear (Pyrus communis L., Cv. Bartlett/William)

    International Nuclear Information System (INIS)

    Gamma-irradiation alone and in combination with refrigeration was tested consecutively for 3 years for extending the shelf life of pear. Matured green pears were irradiated in the dose range of 0.8-2.0 kGy and stored under ambient (temperature 25±2 deg. C, RH 70%) and refrigerated (temperature 3±1 deg. C, RH 80%) conditions. Dose range of 1.5-1.7 kGy extended the storage life of pear by 14 days under ambient conditions. Control unirradiated pears were almost fully ripe within 8 days, while as the pears irradiated in the dose range of 1.5-1.7 kGy were fully ripe within 22 days of ambient storage. Irradiation dose of 1.5-1.7 kGy significantly inhibited the decaying of pears upto 16 days of ambient storage. Irradiation in combination with refrigeration prevented the decaying of pears upto 45 days as against the 35% decay in unirradiated samples. Irradiation dose of 1.5-1.7 kGy also gave an extension of 8 and 4 days during additional ambient storage of the pears following 30 and 45 days of refrigeration, respectively

  17. 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)

  18. 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.

  19. Effect of root pruning and irrigation regimes on leaf water relations and xylem ABA and ionic concentrations in pear trees

    DEFF Research Database (Denmark)

    Wang, Yufei; Bertelsen, Marianne G.; Petersen, Karen Koefoed;

    2014-01-01

    Root pruning is an effective approach for controlling vegetative growth of pear trees (Pyrus communis L.), yet the underlying mechanisms for such effect remain largely elusive. A two-year field experiment was conducted to investigate the effect of root pruning and irrigation regimes on leaf water...... pruning caused water deficit stress in pear trees. Further RP trees had significantly lower concentrations of total cations and anions and the sum of cations and anions than the NP trees implying root pruning decreased acquisition of nutrients from the soil. In the root pruned trees, the leaf water...... as well as in the DI and NI compared to the FI, which could account for the lower stomatal conductance in those treatments. Conclusively, root pruning not only decreased water uptake but also nutrient uptake by the pear trees, and both could have caused reduced vegetative and generative growth...

  20. 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.

  1. Isothermal RNA Sequence Amplification Method for Rapid Antituberculosis Drug Susceptibility Testing of Mycobacterium tuberculosis

    OpenAIRE

    Takakura, Shunji; Tsuchiya, Shigeo; Fujihara, Naoko; Kudo, Toyoichiro; Iinuma, Yoshitsugu; Mitarai, Satoshi; Ichiyama, Satoshi; Yasukawa, Kiyoshi; Ishiguro, Takahiko

    2005-01-01

    RNA transcript quantification by an isothermal sequence amplification reaction was evaluated for susceptibility testing of 15 Mycobacterium tuberculosis strains. Agreement with the proportion method on Ogawa egg medium and the BACTEC MGIT 960 system was 100 and 87% for rifampin, 93 and 100% for isoniazid, 60 and 53% for ethambutol, and 80 and 80% for streptomycin, respectively.

  2. 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

  3. 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

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

    KAUST Repository

    Bukhryakov, Konstantin V.

    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.

  5. 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...

  6. 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.

  7. 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.

  8. [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.

  9. Establishment of growth medium and quantification of pollen grains and germination of pear tree cultivars

    Directory of Open Access Journals (Sweden)

    Paulyene Vieira Nogueira

    2016-06-01

    Full Text Available ABSTRACT To support breeding programs of pear tree, on the selection of cultivars to subtropical area in Brazil, the objective of this research was to adjust the growth medium basic to pollen grain germination. The pollen grains of 'D'Água' cultivar were spread on the surface of Petri dishes containing 20 ml of culture medium in accordance with the following sequential experiments: 1 ágar (4; 6; 8 and 10 g L-1 and pH (3,5; 4,5; 5,5 and 6,5; 2 sucrose (0; 30; 60 and 90 g L-1; 3 calcium nitrate (0; 200; 400 and 800 mg L-1; 4 boric acid (0; 400; 800 and 1200 mg L-1; 5 temperature of incubation (15; 20; 25 e 30 ºC and 6 emission time of the pollen tube (0; 1; 2; 3; 4; 5 and 6 hours after inoculation. After incubation, the germination rate of pollen grains of nine pear cultivars ('Rocha', 'Abate Fetel', 'Packham's Triumph', 'Atago', 'Hosui', 'Primorosa', 'Triunfo', 'Seleta' and 'D'Água' was evaluated, and number of stamens, the number of pollen grains per anther and per flower. The protocol for germination of pollen grains of pear tree consist in the culture medium have to be solidified with 10 g L-1 ágar, being the pH measured to 5,2 , added with 90 g L-1 sucrose, 145 mg L-1 calcium nitrate and 700 mg L-1 boric acid, with incubation temperature of 23 ºC. The readings germination percentage should be performed after five hours of incubation. The pollen grain 'Rocha' cultivar showed higher germination percentage.

  10. 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

  11. 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...

  12. Can Anomalous Amplification be Attained without Postselection?

    Science.gov (United States)

    Martínez-Rincón, Julián; Liu, Wei-Tao; Viza, Gerardo I.; Howell, John C.

    2016-03-01

    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 discarding of data, 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. A tunable phase controls the strength of the amplification response. We experimentally demonstrate the proposed technique by measuring polarization rotations in a linearly polarized laser pulse. We show that in the presence of technical noise the effective sensitivity and precision of a split detector is increased when compared to a conventional continuous-wave balanced detection technique.

  13. 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...

  14. 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.

  15. 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.

  16. 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.

  17. Cascade Signal Amplification Based on Copper Nanoparticle-Reported Rolling Circle Amplification for Ultrasensitive Electrochemical Detection of the Prostate Cancer Biomarker.

    Science.gov (United States)

    Zhu, Ye; Wang, Huijuan; Wang, Lin; Zhu, Jing; Jiang, Wei

    2016-02-01

    An ultrasensitive and highly selective electrochemical assay was first attempted by combining the rolling circle amplification (RCA) reaction with poly(thymine)-templated copper nanoparticles (CuNPs) for cascade signal amplification. As proof of concept, prostate specific antigen (PSA) was selected as a model target. Using a gold nanoparticle (AuNP) as a carrier, we synthesized the primer-AuNP-aptamer bioconjugate for signal amplification by increasing the primer/aptamer ratio. The specific construction of primer-AuNP-aptamer/PSA/anti-PSA sandwich structure triggered the effective RCA reaction, in which thousands of tandem poly(thymine) repeats were generated and directly served as the specific templates for the subsequent CuNP formation. The signal readout was easily achieved by dissolving the RCA product-templated CuNPs and detecting the released copper ions with differential pulse stripping voltammetry. Because of the designed cascade signal amplification strategy, the newly developed method achieved a linear range of 0.05-500 fg/mL, with a remarkable detection limit of 0.020 ± 0.001 fg/mL PSA. Finally, the feasibility of the developed method for practical application was investigated by analyzing PSA in the real clinical human serum samples. The ultrasensitivity, specificity, convenience, and capability for analyzing the clinical samples demonstrate that this method has great potential for practical disease diagnosis applications. PMID:26765624

  18. Onshore seismic amplifications due to bathymetric features

    Science.gov (United States)

    Rodríguez-Castellanos, A.; Carbajal-Romero, M.; Flores-Guzmán, N.; Olivera-Villaseñor, E.; Kryvko, A.

    2016-08-01

    We perform numerical calculations for onshore seismic amplifications, taking into consideration the effect of bathymetric features on the propagation of seismic movements. To this end, the boundary element method is applied. Boundary elements are employed to irradiate waves and, consequently, force densities can be obtained for each boundary element. From this assumption, Huygens’ principle is applied, and since the diffracted waves are built at the boundary from which they are radiated, this idea is equivalent to Somigliana’s representation theorem. The application of boundary conditions leads to a linear system being obtained (Fredholm integral equations). Several numerical models are analyzed, with the first one being used to verify the proposed formulation, and the others being used to estimate onshore seismic amplifications due to the presence of bathymetric features. The results obtained show that compressional waves (P-waves) generate onshore seismic amplifications that can vary from 1.2 to 5.2 times the amplitude of the incident wave. On the other hand, the shear waves (S-waves) can cause seismic amplifications of up to 4.0 times the incident wave. Furthermore, an important result is that in most cases the highest seismic amplifications from an offshore earthquake are located on the shoreline and not offshore, despite the seafloor configuration. Moreover, the influence of the incident angle of seismic waves on the seismic amplifications is highlighted.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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%.

  4. Graft Incompatibility Influence on Assimilating Pigments and Soluble Sugars Amount of some Pear (Pyrus sativa Cultivars

    Directory of Open Access Journals (Sweden)

    Gheorghii CIOBOTARI

    2010-06-01

    Full Text Available Graft incompatibility in fruit trees is one of the greatest obstacles in rootstocks and cultivars breeding. The mechanism in which incompatibility is expressed is not yet fully understood and several hypotheses have been advanced in an attempt to explain it. In many cases (pear on quince grafts, apricot on Prunus grafts, incompatibility is manifested by the breaking of the trees at the point of the union particularly when they have been growing for some years. Many reports focus on this problem in order to understand the mechanisms of graft development. These reports refer to both cytological and biochemical responses occurring at an early phase in response to grafting, as well as to the consequences of these events on the future graft response. In this experiment, we tried to highlight how affinity between scion and rootstock can influence the photosynthetic apparatus and carbohydrates synthesis. The results showed that grafting affinity has an influence on total assimilating pigments content. Thus, on the pear cultivars grafted on an incompatible rootstock (cultivars/Cydonia oblonga the total pigments content ratio (reported to the ungrafted rootstock ranged between 0.58 and 0.69. However, the combinations had a ratio ranging between 0.79 and 0.98. Nevertheless, the assimilating pigments ratio reduction had no influence on photosynthetic rate. The soluble sugars amount was close in both variants (cultivars/Cydonia oblonga and cultivars/Pyrus sativa.

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

    Science.gov (United States)

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

    2012-04-01

    The spiraea aphid (Aphis spiraecola Patch) is a primary pest of fruit trees, particularly pear trees in China. Despite the economic importance of this pest, little is known about its genetic structure or its patterns of dispersal at local and regional scales; however, knowledge of these characteristics is important for establishing effective control strategies for this pest. The genetic variability of 431 individuals from 21 populations on pear trees in China was investigated using eight polymorphic microsatellite loci. The high polymorphism of these markers was evident from the expected heterozygosity value (He = 0.824) and the Polymorphism Information Content (PIC = 0.805), indicating that the spiraea aphid maintains a high level of genetic diversity. The analysis of molecular variance revealed a middle level of population differentiation (F(ST) = 0.1478) among A. spiraecola populations. This result is consistent with the results of the STRUCTURE analysis (K = 3), the unweighted pair-group method with arithmetic average tree and the Mantel test (r = 0.6392; P importance of considering regional differences in studies of population structure, even when strong isolation-by-distance influences the genetic population structure of species. PMID:22606830

  6. 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.

  7. A novel, highly divergent ssDNA virus identified in Brazil infecting apple, pear and grapevine.

    Science.gov (United States)

    Basso, Marcos Fernando; da Silva, José Cleydson Ferreira; Fajardo, Thor Vinícius Martins; Fontes, Elizabeth Pacheco Batista; Zerbini, Francisco Murilo

    2015-12-01

    Fruit trees of temperate and tropical climates are of great economical importance worldwide and several viruses have been reported affecting their productivity and longevity. Fruit trees of different Brazilian regions displaying virus-like symptoms were evaluated for infection by circular DNA viruses. Seventy-four fruit trees were sampled and a novel, highly divergent, monopartite circular ssDNA virus was cloned from apple, pear and grapevine trees. Forty-five complete viral genomes were sequenced, with a size of approx. 3.4 kb and organized into five ORFs. Deduced amino acid sequences showed identities in the range of 38% with unclassified circular ssDNA viruses, nanoviruses and alphasatellites (putative Replication-associated protein, Rep), and begomo-, curto- and mastreviruses (putative coat protein, CP, and movement protein, MP). A large intergenic region contains a short palindromic sequence capable of forming a hairpin-like structure with the loop sequence TAGTATTAC, identical to the conserved nonanucleotide of circoviruses, nanoviruses and alphasatellites. Recombination events were not detected and phylogenetic analysis showed a relationship with circo-, nano- and geminiviruses. PCR confirmed the presence of this novel ssDNA virus in field plants. Infectivity tests using the cloned viral genome confirmed its ability to infect apple and pear tree seedlings, but not Nicotiana benthamiana. The name "Temperate fruit decay-associated virus" (TFDaV) is proposed for this novel virus. PMID:26186890

  8. 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

  9. Heat induces gene amplification in cancer cells

    International Nuclear Information System (INIS)

    Highlights: ► This study discovered that heat exposure (hyperthermia) results in gene amplification in cancer cells. ► Hyperthermia induces DNA double strand breaks. ► DNA double strand breaks are considered to be required for the initiation of gene amplification. ► 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 °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) γH2AX immunostaining to detect γH2AX foci as an indication for DNA double strand breaks. Results: (1) Heat exposure at 42 or 44 °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 °C for 30 min induces DNA double strand breaks in HCT116 cells as shown by γH2AX 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 telomere functions are denatured. To our knowledge, this is the first study to provide direct evidence of hyperthermia induced gene amplification.

  10. 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.

  11. Efficacy of 1-methylcyclopropene on the mitigation of storage disorders of "Rocha" pear under normal refrigerated and controlled atmospheres.

    Science.gov (United States)

    Almeida, Domingos Pf; Carvalho, Rita; Dupille, Eve

    2016-07-01

    Alternatives are needed for long-term preservation of European pears (Pyrus communis L.) after the ban on diphenylamine. "Rocha" pear fruit harvested at commercial maturity were treated with 1-methylcyclopropene (1-methylcyclopropene, SmartFresh™) and diphenylamine and stored at 0 ℃, 90-95% relative humidity, under normal atmosphere for up to six months or under controlled atmosphere (controlled atmosphere, 3 kPa O2 + 0.7 kPa CO2) for up to 9.4 months. At 312 nl l(-1), 1-methylcyclopropene reduced softening and yellowing, and increased soluble solids content during shelf life in comparison with fruit treated with diphenylamine. 1-Methylcyclopropene at 312 nl l(-1) was also more effective than diphenylamine in reducing superficial scald and internal browning disorders. 1-Methylcyclopropene at 150 nl l(-1) had little effect on ripening-related changes but was effective against physiological disorders of pears stored in regular atmosphere or under controlled atmosphere. Delayed controlled atmosphere slightly reduced internal browning disorders but increased superficial scald. 1-Methylcyclopropene at 312 nl l(-1) reduced physiological disorders in "Rocha" pear under refrigerated storage and delayed ripening-related softening and color changes during shelf life. At 150 nl l(-1), 1-methylcyclopropene is as effective as diphenylamine against storage disorders without ripening impairment. PMID:26437671

  12. 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

  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. Metabolic Profiling of Developing Pear Fruits Reveals Dynamic Variation in Primary and Secondary Metabolites, Including Plant Hormones.

    Directory of Open Access Journals (Sweden)

    Akira Oikawa

    Full Text Available Metabolites in the fruits of edible plants include sweet sugars, visually appealing pigments, various products with human nutritional value, and biologically active plant hormones. Although quantities of these metabolites vary during fruit development and ripening because of cell division and enlargement, there are few reports describing the actual dynamics of these changes. Therefore, we applied multiple metabolomic techniques to identify the changes in metabolite levels during the development and ripening of pear fruits (Pyrus communis L. 'La France'. We quantified and classified over 250 metabolites into six groups depending on their specific patterns of variation during development and ripening. Approximately half the total number of metabolites, including histidine and malate, accumulated transiently around the blooming period, during which cells are actively dividing, and then decreased either rapidly or slowly. Furthermore, the amounts of sulfur-containing amino acids also increased in pear fruits around 3-4 months after the blooming period, when fruit cells are enlarging, but virtually disappeared from ripened fruits. Some metabolites, including the plant hormone abscisic acid, accumulated particularly in the receptacle prior to blooming and/or fruit ripening. Our results show several patterns of variation in metabolite levels in developing and ripening pear fruits, and provide fundamental metabolomic data that is useful for understanding pear fruit physiology and enhancing the nutritional traits of new cultivars.

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

    Directory of Open Access Journals (Sweden)

    Oana Viorela NISTOR

    2015-06-01

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

  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. 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.

  19. 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.

  20. Nanguo Pear Management Measures in Autumn and Winter%南果梨秋冬季管理措施

    Institute of Scientific and Technical Information of China (English)

    梁亚华

    2014-01-01

    秋冬季对南果梨进行科学管理是梨果综合管理的一项很重要的内容,它直接影响南果梨等产量、品质及效益。详细阐述秋冬季节南果梨的地下肥水管理、地上病虫防治、修剪等措施,为获得南果梨稳定高产提供参考。%Scientific management of Nanguo pear in the autumn and winter is an important content of comprehensive management of pear, which directly affects the yield, quality and economic benefit of Nanguo pear. This paper gives a detailed description of the under-ground water and fertilizer management, pest control, pruning and other measures in fall and winter seasons in order to provide reference for obtaining high and stable yield of Nanguo pear.

  1. 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.

  2. Development of Wild Birchleaf Pear Fruit Wine%野生棠梨果露酒的研制

    Institute of Scientific and Technical Information of China (English)

    鲍晓华; 毕廷菊; 李秀; 李孙洋

    2011-01-01

    [ Objective ] The research aimed to discuss the development method of wild birchleaf pear fruit wine. [ Method ] Mature fruits of wild birchleaf pear were selected to produce the juice of wild birchleaf pear fruits. And then the juice was mixed with Kaoliang wine by the juice-wine ratio of 2∶1 to develop the wild birchleaf pear fruit wine and its sugar content and alcohol content were measured. [ Result] The alcohol content of the blended wild birchleaf pear fruit wine was 14% - 17% ,pH value was 4.5 and the sugar content was 15% - 18%. The best wine for the preparation of wild birchleaf pear fruit wine was Kaoliang wine and its alcohol content should be 45% -47%. [ Conclusion] In order to ensure the preservation period, the alcohol content of the blended wild birchleaf pear fruit wine should be no less than 15%.%[目的]探索野生棠梨果露酒的研制方法.[方法]挑选成熟野生棠梨果实制成野生棠梨果汁液后,与高粱酒按照汁液:酒为2:1的比例混合均匀,调配成野生棠梨果露酒,测定其糖含量和酒精度.[结果]调配好的野生棠梨果露酒的酒精含量为14%~17%,pH值为4.5,含糖量为15%~18%.用于野生棠梨果露酒调配用的酒最好是高粱酒,酒精含量最好是45%~47%.[结论]为保证野生棠梨果露酒的保存期,调配的野生棠梨果露酒的酒精含量不得低于15%.

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

    Directory of Open Access Journals (Sweden)

    Emmanuelle eFiore

    2015-08-01

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

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. ESTIMATION OF AMPLIFICATION FACTOR IN EARTHQUAKE ENGINEERING

    Directory of Open Access Journals (Sweden)

    Nazarov Yuriy Pavlovich

    2015-03-01

    Full Text Available The authors are the developers of Odyssey Software (Eurosoft Co. for the analysis of seismological data and computing of seismic loads and their parameters. While communicating with the users of the software, the authors have revealed some uncertainty about both understanding of the term "amplification factor (AF" and calculation of the amplification factor using various methods. In this article, a simple example shows that the determination of the amplification factor as the ratio of the acceleration’s spectrum to the maximal acceleration is derived from the classical definition of AF in the form of the ratio of maximal dynamic displacement to the displacement by the action of static load. Deterministic and probabilistic ap-proaches for the calculating of the AF were discussed. There was an example of AFs calculation and their envelopes for translational and rotational components of seismic impact by using Odyssey Software.

  10. 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.

  11. 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.

  12. Radiopolymerization of β(-)pinene: A case of chiral amplification

    International Nuclear Information System (INIS)

    β(-)Pinene was treated with γ 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 β(-)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 β(-)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 β(-)pinene monomer. The results have been discussed in terms of amplification of chirality caused by γ radiation and the implications of this fact on the mechanism of chiral amplification on prebiotic molecules

  13. 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.

  14. 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.

  15. 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.

  16. Ultrasensitive DNA detection based on two-step quantitative amplification on magnetic nanoparticles

    Science.gov (United States)

    Jin, Mingliang; Liu, Xia; van den Berg, Albert; Zhou, Guofu; Shui, Lingling

    2016-08-01

    Sensitive detection of a specific deoxyribo nucleic acid (DNA) sequence is important for biomedical applications. In this report, a two-step amplification strategy is developed based on magnetic nanoparticles (MNPs) to achieve ultrasensitive DNA fluorescence detection. The first level amplification is obtained from multiple binding sites on MNPs to achieve thousands of probe DNA molecules on one nanoparticle surface. The second level amplification is gained by enzymatic reaction to achieve fluorescence signal enhancement. MNPs functionalized by probe DNA (DNAp) are bound to target DNA (t-DNA) molecules with a ratio of 1:1 on a substrate with capture DNA (DNAc). After the MNPs with DNAp are released from the substrate, alkaline phosphatase (AP) is labelled to MNPs via hybridization reaction between DNAp on MNPs and detection DNAs (DNAd) with AP. The AP on MNPs catalyses non-fluorescent 4-methylumbelliferyl phosphate (4-MUP) to fluorescent 4-methylumbelliferone (4-MU) with high intensity. Finally, fluorescence intensity of the 4-MU is detected by a conventional fluorescence spectrophotometer. With this two-step amplification strategy, the limit of detection (LOD) of 2.8 × 10‑18 mol l‑1 for t-DNA has been achieved.

  17. Rapid detection of Mycobacterium tuberculosis by recombinase polymerase amplification.

    Directory of Open Access Journals (Sweden)

    David S Boyle

    Full Text Available Improved access to effective tests for diagnosing tuberculosis (TB has been designated a public health priority by the World Health Organisation. In high burden TB countries nucleic acid based TB tests have been restricted to centralised laboratories and specialised research settings. Requirements such as a constant electrical supply, air conditioning and skilled, computer literate operators prevent implementation of such tests in many settings. Isothermal DNA amplification technologies permit the use of simpler, less energy intensive detection platforms more suited to low resource settings that allow the accurate diagnosis of a disease within a short timeframe. Recombinase Polymerase Amplification (RPA is a rapid, low temperature isothermal DNA amplification reaction. We report here RPA-based detection of Mycobacterium tuberculosis complex (MTC DNA in <20 minutes at 39 °C. Assays for two MTC specific targets were investigated, IS6110 and IS1081. When testing purified MTC genomic DNA, limits of detection of 6.25 fg (IS6110 and 20 fg (IS1081were consistently achieved. When testing a convenience sample of pulmonary specimens from suspected TB patients, RPA demonstrated superior accuracy to indirect fluorescence microscopy. Compared to culture, sensitivities for the IS1081 RPA and microscopy were 91.4% (95%CI: 85, 97.9 and 86.1% (95%CI: 78.1, 94.1 respectively (n = 71. Specificities were 100% and 88.6% (95% CI: 80.8, 96.1 respectively. For the IS6110 RPA and microscopy sensitivities of 87.5% (95%CI: 81.7, 93.2 and 70.8% (95%CI: 62.9, 78.7 were obtained (n = 90. Specificities were 95.4 (95% CI: 92.3,98.1 and 88% (95% CI: 83.6, 92.4 respectively. The superior specificity of RPA for detecting tuberculosis was due to the reduced ability of fluorescence microscopy to distinguish Mtb complex from other acid fast bacteria. The rapid nature of the RPA assay and its low energy requirement compared to other amplification technologies suggest RPA-based TB

  18. 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...

  19. 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. PMID:24650181

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

    Science.gov (United States)

    Butler, P. A.

    2016-07-01

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

  1. 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.

  2. 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.

  3. 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.

  4. Sensitivity to gamma irradiation of post-harvest pathogens of pear

    International Nuclear Information System (INIS)

    Recently, radiation has been used as a fungicidal treatment in the post harvest technology of fruits. The effect of gamma irradiation at doses of 5 - 3000 Gy, on spore germination and mycelial growth of four fungi (Alternaria tenuissima, Botrytis cinerea, Penicillium expansum & Stemphylium botryosum) pathogenic to stored pears were studied. Inhibition of spore germination was found to be directly related to the strength of the radiation dose. B. cinerea and P. expansum were radiation sensitive, while A. tenuissima and S. botryosum were radiation resistant. Exposure of mycelial mat to different radiation doses showed that a dose level of 1000 and 3000 Gy could be considered sufficient for decontamination by the radiosensitive and radio-resistant species, respectively. Regardless of index of mycelial age, young mycelia were more resistant than mature mycelia. The lower doses of gamma radiation increased total proteins and total soluble sugar s of all the tested fungal species but did not effect lipid synthesis

  5. 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.

  6. 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.

  7. A strategy to design efficient fermentation processes for traditional beverages production: prickly pear wine.

    Science.gov (United States)

    Navarrete-Bolaños, J L; Fato-Aldeco, E; Gutiérrez-Moreno, K; Botello-Álvarez, J E; Jiménez-Islas, H; Rico-Martínez, R

    2013-10-01

    This paper describes a methodology to establish an optimal process design for prickly pear wine production that preserves the peculiar and unique traits of traditional products, generating at the same time, technical information for appropriate design of both bioreactor and overall process. The strategy includes alcoholic fermentation optimization by the mixed native culture composed by Pichia fermentans and Saccharomyces cerevisiae, followed by malolactic fermentation optimization by Oenococcus oeni. The optimization criteria were based on multiple output functions: alcohol content, volatile compounds profile, organic acids profile, and compound contents related to color, which were analyzed by spectroscopy-chromatography methods and sensory analysis. The results showed that the mixed culture inoculated into a bioreactor containing prickly pear juice with 20 °Bx of fermentable sugars concentration, processed at a constant temperature of 20 °C for 240 h, leads to a fermented product with 9.93% (v/v) total alcohol content, and significant abundance of volatile compounds, which provide fruity and ethereal aromatic notes, complemented by a lively but not unpleasant acidity. This young wine was further subjected to malolactic fermentation at constant temperature (16 °C) for 192 h, decreasing malic acid, and balancing volatile compounds contents, thus resulting in a product with better aroma and flavor perception, and a velvety feeling of long aftertaste. Repeated assays showed that the process is stable, predictable, controllable, and reproducible. These results were used for process design and spreadsheet construction in order to simulate the process, and properly select and size the equipment required for such process. PMID:24032574

  8. 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.

  9. 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.

  10. Extended alternating-temperature cold acclimation and culture duration improve pear shoot cryopreservation.

    Science.gov (United States)

    Chang, Y; Reed, B M

    2000-06-01

    Meristems of many pear genotypes can be successfully cryopreserved following 1 week of cold acclimation, but an equal number do not survive the process or have very little regrowth. This study compared commonly used cold acclimation protocols to determine whether the cold acclimation technique used affected the cold hardiness of shoots or the regrowth of cryopreserved meristems. In vitro-grown pear (Pyrus L.) shoots were cold acclimated for up to 16 weeks, then either the shoot tips were tested for cold hardiness or the meristems were cryopreserved by controlled freezing. Cold acclimation consisted of alternating temperatures (22 degrees C with light/-1 degrees C darkness with various photo- and thermoperiods) or a constant temperature (4 degrees C with an 8-h photoperiod or darkness). Compared with nonacclimated controls, both alternating- and constant-temperature acclimation significantly improved postcryopreservation regrowth of P. cordata Desv. and P. pashia Buch. -Ham. ex D. Don meristems. Alternating-temperature acclimation combined with either an 8-h photoperiod or darkness was significantly better than constant-temperature acclimation. Alternating-temperature shoot acclimation for 2 to 5 weeks significantly increased postcryopreservation meristem regrowth, and recovery remained high for up to 15 weeks acclimation. Postcryopreservation meristem regrowth increased with 1 to 5 weeks of constant-temperature acclimation and then declined with longer acclimation. Shoot cold hardiness varied with the acclimation procedure. The LT(50) of shoots acclimated for 10 weeks with alternating temperatures was -25 degrees C; that with constant temperature was -14.7 degrees C; and that of the nonacclimated control was -10 degrees C. Less frequent transfer of cultures also improved acclimation of shoots. Shoots grown without transfer to fresh medium for 6-12 weeks had higher postcryopreservation recovery with shorter periods of acclimation than shoots with a 3-week transfer

  11. 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.

  12. 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)).

  13. 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.

  14. 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 ...

  15. Vortical field amplification and particle acceleration at rippled shocks

    CERN Document Server

    Fraschetti, F

    2013-01-01

    Supernova Remnants (SNRs) shocks are believed to accelerate charged particles and to generate strong turbulence in the post-shock flow. From high-energy observations in the past decade, a magnetic field at SNR shocks largely exceeding the shock-compressed interstellar field has been inferred. We outline how such a field amplification results from a small-scale dynamo process downstream of the shock, providing an explicit expression for the turbulence back-reaction to the fluid whirling. The spatial scale of the $X-$ray rims and the short time-variability can be obtained by using reasonable parameters for the interstellar turbulence. We show that such a vortical field saturation is faster than the acceleration time of the synchrotron emitting energetic electrons.

  16. Microchameleons: nonlinear chemical microsystems for amplification and sensing.

    Science.gov (United States)

    Bishop, K J M; Gray, T P; Fialkowski, M; Grzybowski, B A

    2006-09-01

    In biological systems, the coupling of nonlinear biochemical kinetics and molecular transport enables functional sensing and "signal" amplification across many length scales. Drawing on biological inspiration, we describe how artificial reaction-diffusion (RD) microsystems can provide a basis for sensing applications, capable of amplifying micro- and nanoscopic events into macroscopic visual readouts. The RD applications reviewed here are based on a novel experimental technique, WETS for Wet Stamping, which offers unprecedented control over RD processes in microscopic and complex geometries. It is discussed how RD can be used to sense subtle differences in the thickness and/or absorptivity of thin absorptive films, amplify macromolecular phase transitions, detect the presence and quality of self-assembled monolayers, and provide dynamic spatiotemporal readouts of chemical "metabolites." PMID:17014236

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. Optical Pattern Recognition With Self-Amplification

    Science.gov (United States)

    Liu, Hua-Kuang

    1994-01-01

    In optical pattern recognition system with self-amplification, no reference beam used in addressing mode. Polarization of laser beam and orientation of photorefractive crystal chosen to maximize photorefractive effect. Intensity of recognition signal is orders of magnitude greater than other optical correlators. Apparatus regarded as real-time or quasi-real-time optical pattern recognizer with memory and reprogrammability.

  2. Social amplification of risk: a conceptual framework

    International Nuclear Information System (INIS)

    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

  3. Desert Amplification in a Warming Climate

    Science.gov (United States)

    Zhou, Liming

    2016-01-01

    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. PMID:27538725

  4. Desert Amplification in a Warming Climate.

    Science.gov (United States)

    Zhou, Liming

    2016-01-01

    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. PMID:27538725

  5. Electromagnetic waves amplification in a coaxial triode with virtual cathode

    Energy Technology Data Exchange (ETDEWEB)

    Grigoryev, V.P.; Antoshkin, M.Y.; Koval, T.V.; Kuryakov, A.M. [Tomsk Politechnical Univ. (Russian Federation)

    1995-11-01

    The present paper presents the results of analytical and numerical investigations on the amplification of microwaves in the vircator triode of coaxial making. The range of a parameters of the greatest amplification was define for TH and TE-modes.

  6. 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.

  7. Turbulent amplification of magnetic field driven by dynamo effect at rippled shocks

    CERN Document Server

    Fraschetti, Federico

    2013-01-01

    We derive analytically the vorticity generated downstream of a two-dimensional rippled hydromagnetic shock neglecting fluid viscosity and resistivity. The growth of the turbulent component of the downstream magnetic field is driven by the vortical eddies motion. We determine an analytic time-evolution of the magnetic field amplification at shocks, so far described only numerically, until saturation occurs due to seed-field reaction to field lines whirling. The explicit expression of the amplification growth rate and of the non-linear field back-reaction in terms of the parameters of shock and interstellar density fluctuations is derived from MHD jump conditions at rippled shocks. A magnetic field saturation up to the order of milligauss and a short-time variability in the $X$-ray observations of supernova remnants can be obtained by using reasonable parameters for the interstellar turbulence.

  8. Isothermal cycling and cascade signal amplification strategy for ultrasensitive colorimetric detection of nucleic acids

    International Nuclear Information System (INIS)

    We have designed a novel isothermal cascade signal-amplification strategy for ultrasensitive colorimetric determination of nucleic acids. It is based on double-cycling amplification with formation of DNAzyme via a polymerase-induced strand-displacement reaction and nicking endonuclease-assisted recycling. The assay makes use of a hairpin DNA, a short primer, KF-polymerase, and nicking endonuclease. The presence of a target DNA triggers the strand-displacement and polymerization reaction with the formation of numerous DNAzyme molecules. Upon addition of H2O2 to the resulting mixture, the H2O2 reacts with 2,2′-azino-bis (3-ethylbenzothiozoline)-6-sulfonate to form a colored product in the aid of DNAzyme, which is quantified by photometry at 415 nm. Under optimal conditions, the assay allows target DNA to be determined at concentration as low as 0.6 aM. (author)

  9. 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.

  10. Parametric Amplification of Vacuum Fluctuations in a Spinor Condensate

    DEFF Research Database (Denmark)

    Klempt, C.; Topic, O.; Gebreyesus, G.;

    2010-01-01

    Parametric amplification of vacuum fluctuations is crucial in modern quantum optics, enabling the creation of squeezing and entanglement. We demonstrate the parametric amplification of vacuum fluctuations for matter waves using a spinor F=2 87Rb condensate. Interatomic interactions lead to correl......Parametric amplification of vacuum fluctuations is crucial in modern quantum optics, enabling the creation of squeezing and entanglement. We demonstrate the parametric amplification of vacuum fluctuations for matter waves using a spinor F=2 87Rb condensate. Interatomic interactions lead...

  11. 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.

  12. Miniaturized PCR chips for nucleic acid amplification and analysis: latest advances and future trends

    OpenAIRE

    Zhang, Chunsun; Xing, Da

    2007-01-01

    The possibility of performing fast and small-volume nucleic acid amplification and analysis on a single chip has attracted great interest. Devices based on this idea, referred to as micro total analysis, microfluidic analysis, or simply ‘Lab on a chip’ systems, have witnessed steady advances over the last several years. Here, we summarize recent research on chip substrates, surface treatments, PCR reaction volume and speed, architecture, approaches to eliminating cross-contamination and contr...

  13. 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

  14. 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.

  15. 壳聚糖澄清梨汁的研究%Study on clarification of pear juice with chitosan

    Institute of Scientific and Technical Information of China (English)

    付云霄; 张桂

    2012-01-01

    Clarified effect of pear juice using chitosan was studied.Chitosan of concentration 1% was used.After the orthogonal trial,the optimum technological conditions obtained were adding chitosan of 0.2g/L,clarifying for 60min at 35℃,and gained 92.97% transmittance of pear juice under the optimum condition.%采用1%的壳聚糖溶液作用于梨汁,研究了壳聚糖对梨汁的澄清效果。进行正交实验后,获得壳聚糖对梨汁澄清的最适工艺条件:添加0.2g/L壳聚糖,在35℃下澄清60min,梨汁的最高透光率达92.97%。

  16. 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.

  17. MEDICATED PRICKLY PEAR (OPUNTIA FICUS INDICA)-THE NEW EMERGING AGRICULTURAL CROP IN ARID AND SEMI ARID REGIONS OF INDIA

    OpenAIRE

    Chenna Kesava Reddy Sangati; Sucharitha K V; Venkata Ramana D K; Raveendra Reddy Mallela; Syamala B

    2014-01-01

    The present investigation deals with the development of agro-techniques for Opuntia ficus indica (Prickly pear)-OFI cultivation. Standardization and development of best spacing for opuntia plantation, fertilization imposition to achieve good fruit and biomass yield and alternatively fruit quality and biomass parameters was observed and concluded as the better agro-technique among the all imposed treatments. The effect of different spacing and fertilizers composition treatment on cladode yield...

  18. 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. PMID:19096853

  19. 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

    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.%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。

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

    OpenAIRE

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

    2015-01-01

    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...

  1. 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.

  2. Genomic Analysis Reveals a Common Breakpoint in Amplifications of the Plasmodium vivax Multidrug Resistance 1 Locus in Thailand.

    Science.gov (United States)

    Auburn, Sarah; Serre, David; Pearson, Richard D; Amato, Roberto; Sriprawat, Kanlaya; To, Sheren; Handayuni, Irene; Suwanarusk, Rossarin; Russell, Bruce; Drury, Eleanor; Stalker, Jim; Miotto, Olivo; Kwiatkowski, Dominic P; Nosten, Francois; Price, Ric N

    2016-10-15

    In regions of coendemicity for Plasmodium falciparum and Plasmodium vivax where mefloquine is used to treat P. falciparum infection, drug pressure mediated by increased copy numbers of the multidrug resistance 1 gene (pvmdr1) may select for mefloquine-resistant P. vivax Surveillance is not undertaken routinely owing in part to methodological challenges in detection of gene amplification. Using genomic data on 88 P. vivax samples from western Thailand, we identified pvmdr1 amplification in 17 isolates, all exhibiting tandem copies of a 37.6-kilobase pair region with identical breakpoints. A novel breakpoint-specific polymerase chain reaction assay was designed to detect the amplification. The assay demonstrated high sensitivity, identifying amplifications in 13 additional, polyclonal infections. Application to 132 further samples identified the common breakpoint in all years tested (2003-2015), with a decline in prevalence after 2012 corresponding to local discontinuation of mefloquine regimens. Assessment of the structure of pvmdr1 amplification in other geographic regions will yield information about the population-specificity of the breakpoints and underlying amplification mechanisms. PMID:27456706

  3. Penicillium solitum produces a polygalacturonase isozyme in decayed Anjou pear fruit capable of macerating host tissue in vitro.

    Science.gov (United States)

    Jurick, Wayne M; Vico, Ivana; Gaskins, Verneta L; Whitaker, Bruce D; Garrett, Wesley M; Janisiewicz, Wojciech J; Conway, William S

    2012-01-01

    A polygalacturonase (PG) isozyme was isolated from Penicillium solitum-decayed Anjou pear fruit and purified to homogeneity with a multistep process. Both gel filtration and cation exchange chromatography revealed a single PG activity peak, and analysis of the purified protein showed a single band with a molecular mass of 43 kDa, which is of fungal origin. The purified enzyme was active from pH 3.5-6, with an optimum at pH 4.5. PG activity was detectable 0-70 C with 50 C maximum. The purified isozyme was inhibited by the divalent cations Ca(2+), Mg(2+), Mn(2+) and Fe(2+) and analysis of enzymatic hydrolysis products revealed polygalacturonic acid monomers and oligomers. The purified enzyme has an isoelectric point of 5.3 and is not associated with a glycosylated protein. The PG isozyme macerated fruit tissue plugs in vitro and produced ~1.2-fold more soluble polyuronides from pear than from apple tissue, which further substantiates the role of PG in postharvest decay. Data from this study show for the first time that the purified PG produced in decayed Anjou pear by P. solitum, a weakly virulent fungus, is different from that PG produced by the same fungus in decayed apple.

  4. 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%.

  5. 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.

  6. 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.

  7. RNA pre-amplification enables large-scale RT-qPCR gene-expression studies on limiting sample amounts

    Directory of Open Access Journals (Sweden)

    Vermeulen Joëlle

    2009-01-01

    Full Text Available Abstract Background The quantitative polymerase chain reaction (qPCR is a widely utilized method for gene-expression analysis. However, insufficient material often compromises large-scale gene-expression studies. The aim of this study is to evaluate an RNA pre-amplification method to produce micrograms of cDNA as input for qPCR. Findings The linear isothermal Ribo-SPIA pre-amplification method (WT-Ovation; NuGEN was first evaluated by measuring the expression of 20 genes in RNA samples from six neuroblastoma cell lines and of 194 genes in two commercially available reference RNA samples before and after pre-amplification, and subsequently applied on a large panel of 738 RNA samples extracted from neuroblastoma tumours. All RNA samples were evaluated for RNA integrity and purity. Starting from 5 to 50 nanograms of total RNA the sample pre-amplification method was applied, generating approximately 5 microgams of cDNA, sufficient to measure more than 1000 target genes. The results obtained from this study show a constant yield of pre-amplified cDNA independent of the amount of input RNA; preservation of differential gene-expression after pre-amplification without introduction of substantial bias; no co-amplification of contaminating genomic DNA; no necessity to purify the pre-amplified material; and finally the importance of good RNA quality to enable pre-amplification. Conclusion Application of this unbiased and easy to use sample pre-amplification technology offers great advantage to generate sufficient material for diagnostic and prognostic work-up and enables large-scale qPCR gene-expression studies using limited amounts of sample material.

  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. 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.

  10. Diffusive shock acceleration and magnetic field amplification

    CERN Document Server

    Schure, K M; Drury, L O'C; Bykov, A M

    2012-01-01

    Diffusive shock acceleration is the theory of particle acceleration through multiple shock crossings. In order for this process to proceed at a rate that can be reconciled with observations of high-energy electrons in the vicinity of the shock, and for cosmic rays protons to be accelerated to energies up to observed galactic values, significant magnetic field amplification is required. In this review we will discuss various theories on how magnetic field amplification can proceed in the presence of a cosmic ray population. On both short and long scales, cosmic ray streaming can induce instabilities that act to amplify the magnetic field. Developments in this area that have occurred over the past decade are the main focus of this paper.

  11. 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.

  12. 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.

  13. 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

  14. Comparison of nucleic acid hybridization and nucleic acid amplification using conserved sequences from the 5' noncoding region for detection of bovine viral diarrhea virus.

    OpenAIRE

    Ridpath, J F; Bolin, S R; Katz, J

    1993-01-01

    Primers and probes derived from conserved sequences located in the 5' noncoding region of pestiviruses were evaluated for detection of bovine viral diarrhea virus. With these reagents, hybridization and polymerase chain reaction tests detected 62 of 90 and 90 of 90 bovine viral diarrhea virus isolates, respectively. A quick lysis method for preparing RNA for use in polymerase chain reaction amplification also was evaluated.

  15. Introduction to Quantum Noise, Measurement and Amplification

    OpenAIRE

    Clerk, A. A.; Devoret, M. H.; Girvin, S. M.; Marquardt, F.; Schoelkopf, R. J.

    2008-01-01

    The topic of quantum noise has become extremely timely due to the rise of quantum information physics and the resulting interchange of ideas between the condensed matter and AMO/quantum optics communities. This review gives a pedagogical introduction to the physics of quantum noise and its connections to quantum measurement and quantum amplification. After introducing quantum noise spectra and methods for their detection, we describe the basics of weak continuous measurements. Particular atte...

  16. 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.

  17. Non-instrumented nucleic acid amplification assay

    Science.gov (United States)

    Weigl, Bernhard H.; Domingo, Gonzalo; Gerlach, Jay; Tang, Dennis; Harvey, Darrel; Talwar, Nick; Fichtenholz, Alex; van Lew, Bill; LaBarre, Paul

    2008-02-01

    We have developed components of a diagnostic disposable platform that has the dual purpose of providing molecular diagnostics at the point of care (POC) as well as stabilizing specimens for further analysis via a centralized surveillance system. This diagnostic is targeted for use in low-resource settings by minimally trained health workers. The disposable device does not require any additional instrumentation and will be almost as rapid and simple to use as a lateral flow strip test - yet will offer the sensitivity and specificity of nucleic acid amplification tests (NAATs). The low-cost integrated device is composed of three functional components: (1) a sample-processing subunit that generates clean and stabilized DNA from raw samples containing nucleic acids, (2) a NA amplification subunit, and (3) visual amplicon detection sub-unit. The device integrates chemical exothermic heating, temperature stabilization using phase-change materials, and isothermal nucleic acid amplification. The aim of developing this system is to provide pathogen detection with NAAT-level sensitivity in low-resource settings where there is no access to instrumentation. If a disease occurs, patients would be tested with the disposable in the field. A nucleic acid sample would be preserved within the spent disposable which could be sent to a central laboratory facility for further analysis if needed.

  18. 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.

  19. Development of an Automated Microfluidic System for DNA Collection, Amplification, and Detection of Pathogens

    Energy Technology Data Exchange (ETDEWEB)

    Hagan, Bethany S.; Bruckner-Lea, Cynthia J.

    2002-12-01

    This project was focused on developing and testing automated routines for a microfluidic Pathogen Detection System. The basic pathogen detection routine has three primary components; cell concentration, DNA amplification, and detection. In cell concentration, magnetic beads are held in a flow cell by an electromagnet. Sample liquid is passed through the flow cell and bacterial cells attach to the beads. These beads are then released into a small volume of fluid and delivered to the peltier device for cell lysis and DNA amplification. The cells are lysed during initial heating in the peltier device, and the released DNA is amplified using polymerase chain reaction (PCR) or strand displacement amplification (SDA). Once amplified, the DNA is then delivered to a laser induced fluorescence detection unit in which the sample is detected. These three components create a flexible platform that can be used for pathogen detection in liquid and sediment samples. Future developments of the system will include on-line DNA detection during DNA amplification and improved capture and release methods for the magnetic beads during cell concentration.

  20. Ultrasensitive Visual Detection of HIV DNA Biomarkers via a Multi-amplification Nanoplatform.

    Science.gov (United States)

    Long, Yuyin; Zhou, Cuisong; Wang, Congmin; Cai, Honglian; Yin, Cuiyun; Yang, Qiufang; Xiao, Dan

    2016-01-01

    Methodologies to detect disease biomarkers at ultralow concentrations can potentially improve the standard of living. A facile and label-free multi-amplification strategy is proposed for the ultrasensitive visual detection of HIV DNA biomarkers in real physiological media. This multi-amplification strategy not only exhibits a signficantly low detection limit down to 4.8 pM but also provides a label-free, cost-effective and facile technique for visualizing a few molecules of nucleic acid analyte with the naked eye. Importantly, the biosensor is capable of discriminating single-based mismatch lower than 5.0 nM in human serum samples. Moreover, the visual sensing platform exhibits excellent specificity, acceptable reusability and a long-term stability. All these advantages could be attributed to the nanofibrous sensing platform that 1) has a high surface-area-to-volume provided by electrospun nanofibrous membrane, and 2) combines glucose oxidase (GOx) biocatalysis, DNAzyme-catalyzed colorimetric reaction and catalytic hairpin assembly (CHA) recycling amplification together. This multi-amplification nanoplatform promises label-free and visual single-based mismatch DNA monitoring with high sensitivity and specificity, suggesting wide applications that range from virus detection to genetic disease diagnosis. PMID:27032385

  1. 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

  2. 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.

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

    OpenAIRE

    Korenková, Vlasta; Scott, Justin; Novosadová, Vendula; Jindřichová, Marie; Langerová, Lucie; Švec, David; Šídová, Monika; Sjöback, Robert

    2015-01-01

    Background With the introduction of the first high-throughput qPCR instrument on the market it became possible to perform thousands of reactions in a single run compared to the previous hundreds. In the high-throughput reaction, only limited volumes of highly concentrated cDNA or DNA samples can be added. This necessity can be solved by pre-amplification, which became a part of the high-throughput experimental workflow. Here, we focused our attention on the limits of the specific target pre-a...

  4. 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.

  5. 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.

  6. 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. PMID:27284221

  7. G-quadruplex − based homogenous fluorescence platform for ultrasensitive DNA detection through isothermal cycling and cascade signal amplification

    International Nuclear Information System (INIS)

    We describe a simple and homogenous fluorimetric method for sensitive determination of DNA. It is based on target-triggered isothermal cycling and a cascade exponential amplification reaction that generates a large amount of a G-quadruplex. This results in strong fluorescence signal when using thioflavin T as a G-quadruplex-specific light-up fluorescent probe. Tedious handling after amplification is widely eliminated by the addition of thioflavin T. No other exogenous reagent is required. This detection platform is inexpensive and rapid, and displays high sensitivity for target DNA, with a detection limit as low as 91 pM. (author)

  8. 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.

  9. 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. PMID:24210248

  10. 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. PMID:24621296

  11. Effects of the pear tree canopy on photosynthetically active radiation availability

    International Nuclear Information System (INIS)

    The relationships existing between radiant energy and photosynthesis have been extensively investigated on the apple /2/ but not on the other fruit trees, pear included. In addition, such information resists generalization, owing to the remarkable differences underlying tree morphology and physiology of the different species; furthermore, some disagreement arises regarding the terminology and the units used to evaluate the amount of radiant energy useful for the photosynthetic process. In general this evaluation is based on the readouts of illuminance (symbol Ev; unit: lux), in agreement with the photopic curve (fig. 1:A), i.e. with the human eye sensibility to the visible radiation(light). However, the relative response of the chloroplasts to the radiant flux, although included within the same spectral wavebands as the photopic curve, follows a different model (fig.1:B), that is, it has two peaks in connection with the spectral wavelenghts of blue (440–490 nm), and, particularly, of red (620–700 nm). Therefore, according to a number of authors /3/6/8/11/, the correct evaluation of the photosynthetically active radiation should be made using sensors calibrated to measure the photosynthetic photon lux density (symbol: PPFD; unit: μE m-2s-1), and provided with a relative spectral response similar to that of the leaves. (author)

  12. Dickeyafangzhongdai sp. nov., a plant-pathogenic bacterium isolated from pear trees (Pyrus pyrifolia).

    Science.gov (United States)

    Tian, Yanli; Zhao, Yuqiang; Yuan, Xiaoli; Yi, Jianping; Fan, Jiaqin; Xu, Zhigang; Hu, Baishi; De Boer, Solke H; Li, Xiang

    2016-09-01

    Gram-stain-negative, pectinolytic bacteria were repeatedly isolated from pear trees displaying symptoms of bleeding canker in China. Three strains, JS5T, LN1 and QZH3, had identical 16S rRNA gene sequences that shared 99 % similarity to the type strain of Dickeya dadantii. Phylogenetic analysis of strains JS5T, LN1 and QZH3 with isolates representing all species of the genus Dickeya and related Pectobacterium species supported their affiliation to Dickeya. Multi-locus sequence typing employing concatenated sequences encoding recA, fusA, gapA, purA, rplB, dnaX and the intergenic spacer illustrated a phylogeny which placed strains JS5T, LN1 and QZH3 as a distinct clade, separate from all other species of the genus Dickeya. Average nucleotide identity values obtained in comparison with all species of the genus Dickeya supported the distinctiveness of strain JS5T within the genus Dickeya. Additionally, all three strains were phenotypically distinguished from other species of the genus Dickeya by failing to hydrolyse casein, and by producing acids from (-)-d-arabinose, (+)melibiose, (+)raffinose, mannitol and myo-inositol, but not from 5-keto-d-gluconate or β-gentiobiose. The name Dickeya fangzhongdai sp. nov. is proposed to accommodate these strains; the type strain is JS5T (=CGMCC 1.15464T=DSM 101947T). PMID:27045848

  13. Early-type galaxies in the PEARS survey: Probing the stellar populations at moderate redshift

    CERN Document Server

    Ferreras, Ignacio; Malhotra, Sangeeta; Rhoads, James; Cohen, Seth; Windhorst, Rogier; Pirzkal, Nor; Grogin, Norman; Koekkemoer, Anton; Lisker, Thorsten; Panagia, Nino; Daddi, Emanuele; Hathi, Nimish P

    2009-01-01

    Using HST/ACS slitless grism spectra from the PEARS program, we study the stellar populations of morphologically selected early-type galaxies in the GOODS North and South fields. The sample - extracted from a visual classification of the (v2.0) HST/ACS images and restricted to redshifts z>0.4 - comprises 228 galaxies (F775W<24 ABmag) out to z~1.3 over 320 arcmin2, with a median redshift zM=0.75. This work significantly increases our previous sample from the GRAPES survey in the HUDF (18 galaxies over ~11 arcmin2; Pasquali et al. 2006b). The grism data allow us to separate the sample into `red' and `blue' spectra, with the latter comprising 15% of the total. Three different grids of models parameterising the star formation history are used to fit the low-resolution spectra. Over the redshift range of the sample - corresponding to a cosmic age between 5 and 10 Gyr - we find a strong correlation between stellar mass and average age, whereas the **spread** of ages (defined by the RMS of the distribution) is ro...

  14. 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.

  15. 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.

  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. PMID:27537082

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. Parametric Amplification of Gravitational Fluctuations during Reheating

    International Nuclear Information System (INIS)

    Cosmological perturbations can undergo amplification by parametric resonance during preheating even on scales larger than the Hubble radius, without violating causality. A unified description of gravitational and matter fluctuations is crucial to determine the strength of the instability. To extract specific signatures of the oscillating inflaton field during reheating, it is essential to focus on a variable describing metric fluctuations which is constant in the standard analyses of inflation. For a massive inflaton without self-coupling, we find no additional growth of superhorizon modes during reheating beyond the usual predictions. For a massless self-coupled inflaton, there is a sub-Hubble scale resonance. copyright 1999 The American Physical Society

  4. 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.

  5. Novel reverse transcription loop-mediated isothermal amplification for rapid detection of foot-and-mouth disease virus

    DEFF Research Database (Denmark)

    Dukes, J.P.; King, D.P.; Alexandersen, Søren

    2006-01-01

    Speed is paramount in the diagnosis of foot-and-mouth disease (FMD) and simplicity is required if a test is to be deployed in the field. The development of a one-step, reverse transcription loop-mediated amplification (RT-LAMP) assay enables FMD virus (FMDV) to be detected in under an hour...... in a single tube without thermal cycling. A fragment of the 3D RNA polymerase gene of the virus is amplified at 65 degrees C in the presence of a primer mixture and both reverse transcriptase and Bst DNA polymerase. Compared with real-time RT-PCR, RT-LAMP was consistently faster, and ten copies of FMDV...... transcript were detected in twenty-two minutes. Amplification products were detected by visual inspection, agarose gel electrophoresis, or in real-time by the addition of a fluorescent dye. The specificity of the reaction was demonstrated by the absence of amplification of RNA from other viruses that cause...

  6. 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. PMID:26688865

  7. Amplification of 'variola virus-specific' sequences in German cowpox virus isolates.

    Science.gov (United States)

    Meyer, H; Neubauer, H; Pfeffer, M

    2002-02-01

    In 1995 a polymerase chain reaction (PCR) protocol describing the specific detection of variola virus, the causative agent of smallpox, was published by Knight and others. Virulent variola major strains could be differentiated from less virulent variola minor strains because of the distinct amplicon sizes. Here, we applied this PCR protocol to DNA from various orthopoxvirus isolates. There was no amplification with the orthopoxvirus species vaccinia, monkeypox, mousepox, or camelpox viruses. However, amplification was observed in six out of 15 cowpox virus strains investigated. The size of the amplicons corresponded exactly with the size described for variola minor strains and the nucleotide sequence identity accounted for 97%. Findings are discussed with respect to the evolution of orthopoxvirus species assuming that variola virus most probably stems from a rodent-transmitted cowpox virus-like progenitor. PMID:11911586

  8. 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.

  9. Functional integration of PCR amplification and capillary electrophoresis in a microfabricated DNA analysis device.

    Science.gov (United States)

    Woolley, A T; Hadley, D; Landre, P; deMello, A J; Mathies, R A; Northrup, M A

    1996-12-01

    Microfabricated silicon PCR reactors and glass capillary electrophoresis (CE) chips have been successfully coupled to form an integrated DNA analysis system. This construct combines the rapid thermal cycling capabilities of microfabricated PCR devices (10 degrees C/s heating, 2.5 degrees C/s cooling) with the high-speed (Real-time monitoring of PCR target amplification in these integrated PCR-CE devices is also feasible. Amplification of the beta-globin target as a function of cycle number was directly monitored for two different reactions starting with 4 x 10(7) and 4 x 10(5) copies of DNA template. This work establishes the feasibility of performing high-speed DNA analyses in microfabricated integrated fluidic systems. PMID:8946790

  10. 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.

  11. 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.

  12. 复合磷酸盐食品添加剂对鲜切梨保鲜效果的研究%Study on fresh keeping effects of food additives named mixed phosphates on fresh-cut pear

    Institute of Scientific and Technical Information of China (English)

    王丽芳; 王修俊; 郑君花; 李宝升

    2013-01-01

    鲜切梨在加工过程中易使酶和酚类物质在有氧气的条件下相互接触发生酶促褐变,从而严重影响梨的颜色、风味和品质;同时,叶绿素在食品加工中可发生脱镁反应,生成暗绿色至绿褐色的脱镁叶绿素,从而影响鲜切梨的色泽.由于使用单组分的保鲜剂,很难在控制褐变的同时护色保绿.因此本实验将三聚磷酸钠(STP)、L-半胱氨酸(L-cys)和氯化钠(NaCl)食品添加剂制成复合添加剂,将其应用到鲜切梨的抗褐变、保鲜中,利用复合添加剂中各组分的协同和互补作用,既能防止鲜切梨的酶促褐变又能解决叶绿素褐变的问题,从而延长鲜切梨的保藏期.结果表明:复合食品添加剂的最优组合为0.20%三聚磷酸钠,0.05%L-半胱氨酸和0.25%氯化钠,此时鲜切梨的抗褐变效果最好,保鲜效果最佳.%The fresh-cut pears occur to the enzyme browning easily if the enzyme and the phenolic compounds touch each other under the condition of oxygen which seriously affect the color,flavor and quality of pear.At the same time,chlorophyll can occur to the takeing-off magnesium reaction in the food processing,generating dark green pheophytin to greenish-brown pheophytin,which affect the colour and lustre of fresh-cut pears.Due to the use of single-component antistaling agent,it is difficult to control the enzyme browning and protect green color.This thesis studied that the compound reagent of the sodium tripolyphosphate (STP),L-cys and NaCl are compounded and applied to the fresh-cut pears.It can not only prevent effectively the enzyme browning,but also solve the chlorophyll browning problem utilizing the function of cooperating with other component.The results indicated that the best combined ratio of mixed phosphates was 0.2% STP,0.05% L-cys and 0.25% NaCl and the result was the best for fresh-keeping.

  13. Amplification of fluorescence using collinear picosecond optical parametric amplification at degeneracy

    Institute of Scientific and Technical Information of China (English)

    Zhang Jing; Zhang Qiu-Lin; Jiang Man; Zhang Dong-Xiang; Feng Bao-Hua; Zhang Jing-Yuan

    2012-01-01

    We demonstrate the output characteristic of broadband parametric amplification of incoherent light pulses in a 355-nm pumped degenerate picosecond optical parametric amplification with either saturated or unsaturated amplification.The optical parametric amplifier is seeded by the fluorescence generated in a solution of pyridine-1 dye in ethanol.With the saturated amplification,we can obtain high energy incoherent light pulses,whose full widtth at half maximum bandwidth varies from 16 nm to 53 nm for the different phase matching angles near degeneracy.Moreover,the unsaturated bandwidth of the amplified pulses fits well to the calculated result at degeneracy.Selecting s-polarized fluorescence with a Glan-Taylor prism,the maximum bandwidth of the amplified fluorescence is found to be 59 nm for a purely s-polarized seed.The maximum output energy is 0.67 mJ for the optical parametric amplifier.By using an optical filter and compressor,the generated high energy incoherent light has great potential as the incoherent pump,signal or idler wave of a parametric down-conversion process,so that a wave with a high degree of coherence can be generated from an incoherent pump light.

  14. Multiplex allele-specific target amplification based on PCR suppression

    OpenAIRE

    Broude, Natalia E.; Zhang, Lingang; Woodward, Karen; Englert, David; Cantor, Charles R.

    2001-01-01

    We have developed a strategy for multiplex PCR based on PCR suppression. PCR suppression allows DNA target amplification with only one sequence-specific primer per target and a second primer that is common for all targets. Therefore, an n-plex PCR would require only n + 1 primers. We have demonstrated uniform, efficient amplification of targeted sequences in 14-plex PCR. The high specificity of suppression PCR also provides multiplexed amplification with allele specifi...

  15. Loss of KLF14 triggers centrosome amplification and tumorigenesis

    OpenAIRE

    Fan, Guangjian; Sun, Lianhui; Shan, Peipei; Zhang, Xianying; Huan, Jinliang; Li, Dali; Wang, Tingting; Wei, Tingting; Zhang, Xiaohong; Gu, Xiaoyang; Yao, Liangfang; Xuan, Yang; Hou, Zhaoyuan; Cui, Yongping; Cao, Liu

    2015-01-01

    Centrosome amplification is frequent in cancer, but the underlying mechanisms remain unclear. Here we report that disruption of the Kruppel-like factor 14 (KLF14) gene in mice causes centrosome amplification, aneuploidy and spontaneous tumorigenesis. Molecularly, KLF14 functions as a transcriptional repressor of Plk4, a polo-like kinase whose overexpression induces centrosome overduplication. Transient knockdown of KLF14 is sufficient to induce Plk4-directed centrosome amplification. Clinical...

  16. 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.

  17. Effects of aortic root motion on wall stress in the Marfan aorta before and after personalised aortic root support (PEARS) surgery.

    Science.gov (United States)

    Singh, S D; Xu, X Y; Pepper, J R; Izgi, C; Treasure, T; Mohiaddin, R H

    2016-07-01

    Aortic root motion was previously identified as a risk factor for aortic dissection due to increased longitudinal stresses in the ascending aorta. The aim of this study was to investigate the effects of aortic root motion on wall stress and strain in the ascending aorta and evaluate changes before and after implantation of personalised external aortic root support (PEARS). Finite element (FE) models of the aortic root and thoracic aorta were developed using patient-specific geometries reconstructed from pre- and post-PEARS cardiovascular magnetic resonance (CMR) images in three Marfan patients. The wall and PEARS materials were assumed to be isotropic, incompressible and linearly elastic. A static load on the inner wall corresponding to the patients' pulse pressure was applied. Cardiovascular MR cine images were used to quantify aortic root motion, which was imposed at the aortic root boundary of the FE model, with zero-displacement constraints at the distal ends of the aortic branches and descending aorta. Measurements of the systolic downward motion of the aortic root revealed a significant reduction in the axial displacement in all three patients post-PEARS compared with its pre-PEARS counterparts. Higher longitudinal stresses were observed in the ascending aorta when compared with models without the root motion. Implantation of PEARS reduced the longitudinal stresses in the ascending aorta by up to 52%. In contrast, the circumferential stresses at the interface between the supported and unsupported aorta were increase by up to 82%. However, all peak stresses were less than half the known yield stress for the dilated thoracic aorta. PMID:27255604

  18. Characterization of CIPK Family in Asian Pear (Pyrus bretschneideri Rehd) and Co-expression Analysis Related to Salt and Osmotic Stress Responses

    Science.gov (United States)

    Tang, Jun; Lin, Jing; Li, Hui; Li, Xiaogang; Yang, Qingsong; Cheng, Zong-Ming; Chang, Youhong

    2016-01-01

    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.

  19. 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.

  20. 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.

  1. Space Optical Communications Using Laser Beam Amplification

    Science.gov (United States)

    Agrawal, Govind

    2015-01-01

    The Space Optical Communications Using Laser Beam Amplification (SOCLBA) project will provide a capability to amplify a laser beam that is received in a modulating retro-reflector (MRR) located in a satellite in low Earth orbit. It will also improve the pointing procedure between Earth and spacecraft terminals. The technology uses laser arrays to strengthen the reflected laser beam from the spacecraft. The results of first year's work (2014) show amplification factors of 60 times the power of the signal beam. MMRs are mirrors that reflect light beams back to the source. In space optical communications, a high-powered laser interrogator beam is directed from the ground to a satellite. Within the satellite, the beam is redirected back to ground using the MMR. In the MMR, the beam passes through modulators, which encode a data signal onto the returning beam. MMRs can be used in small spacecraft for optical communications. The SOCLBA project is significant to NASA and small spacecraft due to its application to CubeSats for optical data transmission to ground stations, as well as possible application to spacecraft for optical data transmission.

  2. Optimization of noncollinear optical parametric amplification

    Science.gov (United States)

    Schimpf, D. N.; Rothardt, J.; Limpert, J.; Tünnermann, A.

    2007-02-01

    Noncollinearly phase-matched optical parametric amplifiers (NOPAs) - pumped with the green light of a frequency doubled Yb-doped fiber-amplifier system 1, 2 - permit convenient generation of ultrashort pulses in the visible (VIS) and near infrared (NIR) 3. The broad bandwidth of the parametric gain via the noncollinear pump configuration allows amplification of few-cycle optical pulses when seeded with a spectrally flat, re-compressible signal. The short pulses tunable over a wide region in the visible permit transcend of frontiers in physics and lifescience. For instance, the resulting high temporal resolution is of significance for many spectroscopic techniques. Furthermore, the high magnitudes of the peak-powers of the produced pulses allow research in high-field physics. To understand the demands of noncollinear optical parametric amplification using a fiber pump source, it is important to investigate this configuration in detail 4. An analysis provides not only insight into the parametric process but also determines an optimal choice of experimental parameters for the objective. Here, the intention is to design a configuration which yields the shortest possible temporal pulse. As a consequence of this analysis, the experimental setup could be optimized. A number of aspects of optical parametric amplifier performance have been treated analytically and computationally 5, but these do not fully cover the situation under consideration here.

  3. 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 ...

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

    Science.gov (United States)

    Tougan, Takahiro; Okuzaki, Daisuke; Nojima, Hiroshi

    2008-09-01

    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 synthesis from only 0.49 femtograms of mRNA (730 mRNA molecules) as a substrate, a quantity that corresponds to a minor population of mRNA molecules in a single mammalian cell. Analysis of the independent cDNA clone of this library (6.6 x 10(5) cfu) suggests that 30-fold RNA amplification occurred in each round of the amplification process. The size distribution and representation of mRNAs in the resulting one-cell cDNA library retained its similarity to that of the million-cell cDNA library. The use of chum-RNA might also facilitate reactions involving other DNA/RNA modifying enzymes whose Michaelis constant (K(m)) values are around 1 mM, allowing them to be activated in the presence of only small quantities of substrate. PMID:18603591

  5. Non-destructive Measurement of Calcium and Potassium in Apple and Pear Using Handheld X-ray Fluorescence.

    Science.gov (United States)

    Kalcsits, Lee A

    2016-01-01

    Calcium and potassium are essential for cell signaling, ion homeostasis and cell wall strength in plants. Unlike nutrients such as nitrogen and potassium, calcium is immobile in plants. Localized calcium deficiencies result in agricultural losses; particularly for fleshy horticultural crops in which elemental imbalances in fruit contribute to the development of physiological disorders such as bitter pit in apple and cork spot in pear. Currently, elemental analysis of plant tissue is destructive, time consuming and costly. This is a limitation for nutrition studies related to calcium in plants. Handheld portable x-ray fluorescence (XRF) can be used to non-destructively measure elemental concentrations. The main objective was to test if handheld XRF can be used for semi-quantitative calcium and potassium analysis of in-tact apple and pear. Semi-quantitative measurements for individual fruit were compared to results obtained from traditional lab analysis. Here, we observed significant correlations between handheld XRF measurements of calcium and potassium and concentrations determined using MP-AES lab analysis. Pearson correlation coefficients ranged from 0.73 and 0.97. Furthermore, measuring apple and pear using handheld XRF identified spatial variability in calcium and potassium concentrations on the surface of individual fruit. This variability may contribute to the development of localized nutritional imbalances. This highlights the importance of understanding spatial and temporal variability in elemental concentrations in plant tissue. Handheld XRF is a relatively high-throughput approach for measuring calcium and potassium in plant tissue. It can be used in conjunction with traditional lab analysis to better understand spatial and temporal patterns in calcium and potassium uptake and distribution within an organ, plant or across the landscape. PMID:27092160

  6. 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.

  7. 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.

  8. Evaluation of two drip irrigation systems in production and fruit quality of pear (pyrus communis l.) cv. triunfo de viena

    OpenAIRE

    Arenas Bautista, María Cristina; Vélez Sánchez, Javier Enrique; Camacho Tamayo, Jesús Hernán

    2012-01-01

    The rational use of water becomes every day ever more important in fruit production. The aim of this study was to evaluate the effect of the use of double drip line from a line by line plant in pear (Pyrus communis) production, cv. Triunfo de Viena. The research was based on the application of two specific processes, one consisting of a drip line per row of plants with six emitters of 8 lt h-1 and another with two drip lines per row of plants with three emitters of 8 lt h-1 each. Watermark se...

  9. Foraging behavior and pollinating effectiveness of Osmia cornuta (Hymenoptera: Megachilidae) and Apis mellifera (Hymenoptera: Apidae) on "Comice" pear

    OpenAIRE

    Monzón, Víctor; Bosch, Jordi; Retana, Javier

    2004-01-01

    We studied the pollinating effectiveness of Osmia cornuta and Apis mellifera on 'Comice' pear. Osmia cornuta visited more flowers per minute (13.8) than A. mellifera (7.1-9.8). Both species visited similar numbers of flowers per tree (6.7-7.9), and switched rows with similar frequency (4.0-7.9%). Rate of stigma contact was 98.7% for O. cornuta, 51.8% for A. mellifera pollen-nectar foragers, and 19.0% for A. mellifera nectar foragers. Fruit-set in flowers visited once was 28.9, 29.3, and 12.9%...

  10. Amp-PCR: Combining a Random Unbiased Phi29-Amplification with a Specific Real-Time PCR, Performed in One Tube to Increase PCR Sensitivity

    OpenAIRE

    Lena Erlandsson; Lars Peter Nielsen; Anders Fomsgaard

    2010-01-01

    In clinical situations where a diagnostic real-time PCR assay is not sensitive enough, leading to low or falsely negative results, or where detection earlier in a disease progression would benefit the patient, an unbiased pre-amplification prior to the real-time PCR could be beneficial. In Amp-PCR, an unbiased random Phi29 pre-amplification is combined with a specific real-time PCR reaction. The two reactions are separated physically by a wax-layer (AmpliWax®) and are run in sequel in the sam...

  11. Conservation of apple and pear juice concentrates. Synergic effect of heat and radiation

    International Nuclear Information System (INIS)

    This paper aims at assessing the feasibility for conserving apple and pear-juice concentrates through the synergic action of heat and radiation. The material was packed in sterile 100-μm polyethylene bags and, after the treatment was applied, the resulting fractions were stored under room temperature (250C+-10C). The temperature applied to the samples before irradiation was 500C during 10 minutes and the doses were 100, 200, 300 and 400 krad. For such purposes, a 60Co 165-krad/h source was used, located in a mobile irradiator. Periodical microbiological, chemical and organoleptic controls of the food were performed on both control and on irradiated samples, with or without heat. A single alterating microorganism was isolated from all the samples, which was featured as Saccharomyces rouxii. Adopting the temperature as an application variable and the absorbed dose as a constant, the above osmophilic yeast is considerably more sensitive to radiations when it is suspended in a 50% sucrose solution, after the latter was submitted to a 500C temperature treatment. It has been proved that 72.5 krad are needed to attain the reduction of a logarithmic cycle in the Saccharomyces rouxii population irradiated at room temperature, while 36 krad are needed if the sample has been previously heated to 500C for 10 minutes. An attempt was made to apply the synergism of such process to the juice concentrate. Below 500C associated with 400 krad gamma radiation, a total inactivation took place in the Saccharomyces rouxii during the 150 days under analysis. Colour changes were detected in the concentrate; however, the acceptability features for consumption remained at a normal value (level 5) in a hedonic scale of 7 points. (author)

  12. 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.

  13. Promotion of Flowering by Apple Latent Spherical Virus Vector and Virus Elimination at High Temperature Allow Accelerated Breeding of Apple and Pear.

    Science.gov (United States)

    Yamagishi, Norioko; Li, Chunjiang; Yoshikawa, Nobuyuki

    2016-01-01

    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 2 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 3 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 4-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 combining the promotion

  14. 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

  15. A new ultrasonic signal amplification method for detection of bacteria

    International Nuclear Information System (INIS)

    A new method is presented that increases the sensitivity of ultrasound-based techniques for detection of bacteria. The technique was developed for the detection of catalase-positive microorganisms. It uses a bubble trapping medium containing hydrogen peroxide that is mixed with the sample for microbiological evaluation. The enzyme catalase is present in catalase-positive bacteria, which induces a rapid hydrolysis of hydrogen peroxide, forming bubbles which remain in the medium. This reaction results in the amplification of the mechanical changes that the microorganisms produce in the medium. The effect can be detected by means of ultrasonic wave amplitude continuous measurement since the bubbles increase the ultrasonic attenuation significantly. It is shown that microorganism concentrations of the order of 105 cells ml−1 can be detected using this method. This allows an improvement of three orders of magnitude in the ultrasonic detection threshold of microorganisms in conventional culture media, and is competitive with modern rapid microbiological methods. It can also be used for the characterization of the enzymatic activity. (paper)

  16. 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.)

  17. Signal Amplification of Bioassay Using Zinc Nanomaterials

    Science.gov (United States)

    Cowles, Chad L.

    An emerging trend in the analytical detection sciences is the employment of nanomaterials for bioassay signal transduction to identify analytes critical to public health. These nanomaterials have been specifically investigated for applications which require identification of trace levels of cells, proteins, or other molecules that can have broad ranging impacts to human health in fields such as clinical diagnostics, environmental monitoring, food and drink control, and the prevention of bioterrorism. Oftentimes these nanoparticle-based signal transduction or amplification approaches offer distinct advantages over conventional methods such as increased sensitivity, rapidity, or stability. The biological application of nanoparticles however, does suffer from drawbacks that have limited more widespread adoption of these techniques. Some of these drawbacks are, high cost and toxicity, arduous synthesis methods, functionalization and bioconjugation challenges, and laboratory disposal and environmental hazard issues, all of which have impeded the progression of this technology in some way or another. This work aims at developing novel techniques that offer solutions to a number of these hurdles through the development of new nanoparticle-based signal transduction approaches and the description of a previously undescribed nanomaterial. Zinc-based nanomaterials offer the opportunity to overcome some of the limitations that are encountered when other nanomaterials are employed for bioassay signal transduction. On the other hand, the biological application of zinc nanomaterials has been difficult because in general their fluorescence is in the blue range and the reported quantum yields are usually too low for highly sensitive applications. The advantages of using zinc nanomaterials for biological applications, such as reduced toxicity, simple synthesis, low cost, and straightforward functionalization strategies contribute to the research interest in their application as

  18. 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

  19. 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...

  20. 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.

  1. 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.

  2. Strengthening weak value amplification with recycled photons

    CERN Document Server

    Dressel, Justin; Jordan, Andrew N; Graham, Trent M; Kwiat, Paul G

    2013-01-01

    We consider the use of cyclic weak measurements to improve the sensitivity of weak-value amplification precision measurement schemes. Previous weak-value experiments have used only a small fraction of events, while discarding the rest through the process of "post-selection". We extend this idea by considering recycling of events which are typically unused in a weak measurement. Here we treat a sequence of polarized laser pulses effectively trapped inside an interferometer using a Pockels cell and polarization optics. In principle, all photons can be post-selected, which will improve the measurement sensitivity. We first provide a qualitative argument for the expected improvements from recycling photons, followed by the exact result for the recycling of collimated beam pulses, and numerical calculations for diverging beams. We show that beam degradation effects can be mitigated via profile flipping or Zeno reshaping. The main advantage of such a recycling scheme is an effective power increase, while maintainin...

  3. 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.

  4. Beyond the diffraction limit via optical amplification

    CERN Document Server

    Kellerer, Aglae N

    2016-01-01

    In a previous article we suggested a method to overcome the diffraction limit behind a telescope. We refer to theory and recent numerical simulations, and test whether it is indeed possible to use photon amplification to enhance the angular resolution of a telescope or a microscope beyond the diffraction limit. An essential addition is the proposal to select events with above-average ratio of stimulated to spontaneous photons. We find that the diffraction limit of a telescope is surpassed by a factor ten for an amplifier gain of 200, if the analysis is restricted to a tenth of the incoming astronomical photons. A gain of 70 is sufficient with a hundredth of the photons.

  5. 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.

  6. 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.

  7. The Quantum Theory of Optical Parametric Amplification

    Science.gov (United States)

    Hussain, N. A.

    Available from UMI in association with The British Library. Requires signed TDF. The aim of this thesis is to investigate the effect of parametric amplification on various forms of light. In particular we shall consider number and coherent states, but many of the calculations hold for those states whose operators satisfy the properties, = = ==0 e.g. chaotic light. The first chapter lays down the fundamental preliminaries necessary for our calculations and reviews linear amplifier theory. We consider the phase sensitive and insensitive forms of amplifiers modelling the former on the degenerate parametric amplifier and the latter on the non-degenerate and inverted population amplifiers. Chapter 2 deals with balanced homodyne detection of a narrow band coherent state before and after degenerate parametric amplification. In chapter 3 we consider a continuous mode number state produced by atomic emission and parametrically amplified using the formalism of Collett and Gardiner. We give general results for the output flux intensity and also consider the simpler case where the atomic decay rate is much smaller than the parametric cavity decay rate. Also we consider the degree of second order coherence using this simplified theory. Chapters 4 and 5 consider the double amplifier interferometer, using single and continuous mode theories, and enable us to determine the form of amplifier which produces the best visibility and hence lowest noise figures. The travelling-wave parametric amplifier is discussed in chapter 6 and is contrasted with the cavity parametric amplifier discussed in chapters 1 and 2. Finally we consider the much contemplated idea of using amplifiers to boost signals in fibre optic transmission lines using our model of the parametric amplifier and examining the degradation of the signal-to-noise ratio. We consider both coherent and squeezed inputs and our results hold for both cavity and travelling -wave amplifiers.

  8. The Shortwave Infrared Bands’ Response to Stomatal Conductance in “Conference” Pear Trees (Pyrus communis L.

    Directory of Open Access Journals (Sweden)

    Raymond Struthers

    2015-10-01

    Full Text Available In situ measurements consisting of stomatal conductance, air temperature, vapor pressure deficit and the spectral reflectance in the shortwave infrared (SWIR regions of thirty “Conference” pear trees (Pyrus communis L. were repeatedly measured for eighty-six days. The SWIR was segmented into eight regions between 1550 and 2365 nm, where distances ranged from 40–200 nm. Each of the regions was used to describe the change in canopy water status over a period of approximately three months. Stomatal conductance of the water stress treatment was first determined to be significantly different from the control group nine days after stress initiation. The most suitable SWIR region for this study had wavelengths between 1550 and 1750 nm, where the first significant difference was also measured nine days after stress was initiated. After the period of water stress ended, forty-seven days after stress was initiated, all of the trees received full irrigation, where the SWIR region between 1550 and 1750 nm determined that stomatal conductance of the stress treatment lagged behind the control group for thirty days. Using a temporal sequence of SWIR measurements, we were able to successfully measure the beginning and the recovery of water stress in pear trees.

  9. Asynchronous ripening behavior of cactus pear (Opuntia ficus-indica) cultivars with respect to physicochemical and physiological attributes.

    Science.gov (United States)

    Kyriacou, M C; Emmanouilidou, M G; Soteriou, G A

    2016-11-15

    Physicochemical and physiological ripening events in cactus pear (Opuntia ficus-indica) fruit of cultivars 'Ntopia' and 'Hercules' were profiled against skin coloration from mature-green (S1) to over-mature (S5). Fructose and glucose accumulation were linear in 'Ntopia' but peaked near S3 in 'Hercules' synchronously to the appearance of sucrose. Betalains increased steadily in 'Ntopia' (103.2mg/l) but peaked before full skin coloration in 'Hercules' (49.7mg/l); whereas phenolic content remained invariable and ascorbate content peaked near S5 in both 'Ntopia' (108.6μg/g) and 'Hercules' (163.1μg/g). Cell wall material diminished with maturity though textural changes with ripening appeared not related to pectin solubilization but to weakening of glycan bonding and loss of neutral sugars. Fruit firmness rather was correlated to seed weight (r=0.89) and seed-to-pulp ratio (r=0.73). Cultivar differences highlighted in the chronology of ripening events are critical for defining optimum harvest maturity and postharvest handling protocols for premium quality cactus pear fruit. PMID:27283673

  10. Determination of plant growth regulators in pears by microwave-assisted extraction and liquid chromatography with electrospray ionization mass spectrometry.

    Science.gov (United States)

    Mao, Xuejin; Tang, Lijuan; Tan, Ting; Wan, Yiqun

    2014-06-01

    A new method for the determination of six plant growth regulators, 3-indolylacetic acid, 3-indolepropionic acid, 2-naphthoxyacetic acid, 2,4-dicholrophenoxyacetic acid, 1-naphthlcetic acid, and methyl naphthalene-1-acetate, in pears was established by liquid chromatography with electrospray ionization mass spectrometry. In this study, a microwave-assisted extraction technique was first applied for the determination of plant growth regulators in fruit and three cleanup techniques were, respectively, investigated for the purification of pear samples. The chromatographic separation was performed on a Diamonsil C18 column by using 0.01 mol/L formic acid/ammonium formate buffer solution (pH 3.5)/methanol (35:65, v/v) as the mobile phase with a flow rate of 0.7 mL/min in 1:1 split mode. The LODs ranged from 0.3 to 1.9 μg/kg. Under optimized conditions, the average recoveries (five replicates) for six plant growth regulators (spiked at 0.01, 0.05, and 0.5 mg/kg) ranged from 78.9 to 118.0%, and the RSDs were 1.4-10.3%.

  11. Dormancy of 'Imperial Gala' apple and 'Hosui' pear tree buds in a region of low chill occurrence

    Directory of Open Access Journals (Sweden)

    Ruy Inácio Neiva de Carvalho

    2014-08-01

    Full Text Available The objective of this work was to evaluate the dormancy dynamic of Imperial Gala apple tree buds and Hosui pear tree buds in a region of low chill occurrence. Experiments were conducted between April and August in 2007 and 2008. Branches were collected every two weeks from an orchard at Porto Amazonas (Paraná State, Brazil. On the last sampling day, an additional set of branches was collected and refrigerated between 4°C and 7°C for 1,440 hours. Dormancy was evaluated using a biological test of single node cuttings isolated in growth chambers (GC at 25°C with 16 hours of light exposure. The number of chill hours (CH and chill units (CU for the region were recorded. The two species were evaluated in separate experiments. We used 11 completely randomized treatments with four replicas for each species. The peak of endodormancy for the Imperial Gala apple tree buds occurred in early June 2007 and from middle June to early July in 2008. The endodormancy of the Hosui pear tree buds oscillated between April and August in 2007 and peaked between June and early July in 2008.

  12. Isolation, Purification, and Characterization of an Endogenous Root-promoting Factor Obtained from Basal Sections of Pear Hardwood Cuttings.

    Science.gov (United States)

    Fadl, M S; Hartmann, H T

    1967-04-01

    Basal segments taken from Old Home and Bartlett pear hardwood cuttings collected at intervals during the rooting period in September were extracted with ethanol and fractionated by paper chromatography in different solvent systems. Different zones on the chromatograms were bioassayed by the mung bean rooting test, which showed high levels of promotion in Old Home basal extracts when the cuttings were obtained during the period of maximum rooting. Extracts from Bartlett cuttings, however, showed considerably less promotion activity in the bioassay but did show high levels of inhibitory activity.After the easily-rooted Old Home cuttings had been in the rooting medium for 10 days, a highly active endogenous root-promoting material was found in extracts from basal segments of cuttings having buds and which had been treated with indolebutyric acid. Similar extracts obtained from disbudded cuttings, or from cuttings with buds but not treated with indolebutyric acid, lacked this rooting-factor. Extracts obtained from all types of the difficult-to-root Bartlett cuttings also lacked this rooting-factor. The latter is believed to be produced by physiologically active Old Home buds, and is very effective in the mung bean bioassay, even at extremely low concentrations.From paper chromatographic studies, tests with spray reagents, solubility determinations, biological tests, UV spectrum analysis, and infrared spectroscopy, it is believed that this rooting factor could be a condensation product between exogenous auxin (indolebutyric acid) and a phenolic compound produced by physiologically active Old Home pear buds. PMID:16656535

  13. Prodigious polyphyly in imperilled freshwater pearly-mussels (Bivalvia: Unionidae): a phylogenetic test of species and generic designations

    Science.gov (United States)

    2000-01-01

    Unionid bivalves or freshwater pearly-mussels (Unionoidea: Unionidae) serve as an exemplary system for examining many of the problems facing systematists and conservation biologists today. Most of the species and genera were described in the late 1800s and early 1900s, but few phylogenetic studies have been conducted to test conventional views of species and classification. Pearly-mussels of Gulf Coastal drainages of the southeastern United States from the Escambia (southern Alabama to Florida) to the Suwannee Rivers (Florida) are a unique fauna comprised of approximately 100 species, with about 30 endemic to the region. In this study, mitochondrial cytochrome c oxidase subunit I and 16S rRNA gene sequences were used to test the monophyly and to estimate evolutionary relationships of five unionid species representing three different genera. The molecular phylogenies depict all three genera as polyphyletic. The prodigious polyphyly exhibited within unionids is due to incorrect notions of homology and false assumptions about missing anatomical data. In contrast, the molecular phylogeny provides evidence to support the recognition of all five unionid species as distinct evolutionary entities. Furthermore, molecular genealogical evidence supports the elevation of Quincuncina infucata (Conrad) of the Suwannee River to species level, for which Q. kleiniana (Lea) is available.

  14. Asynchronous ripening behavior of cactus pear (Opuntia ficus-indica) cultivars with respect to physicochemical and physiological attributes.

    Science.gov (United States)

    Kyriacou, M C; Emmanouilidou, M G; Soteriou, G A

    2016-11-15

    Physicochemical and physiological ripening events in cactus pear (Opuntia ficus-indica) fruit of cultivars 'Ntopia' and 'Hercules' were profiled against skin coloration from mature-green (S1) to over-mature (S5). Fructose and glucose accumulation were linear in 'Ntopia' but peaked near S3 in 'Hercules' synchronously to the appearance of sucrose. Betalains increased steadily in 'Ntopia' (103.2mg/l) but peaked before full skin coloration in 'Hercules' (49.7mg/l); whereas phenolic content remained invariable and ascorbate content peaked near S5 in both 'Ntopia' (108.6μg/g) and 'Hercules' (163.1μg/g). Cell wall material diminished with maturity though textural changes with ripening appeared not related to pectin solubilization but to weakening of glycan bonding and loss of neutral sugars. Fruit firmness rather was correlated to seed weight (r=0.89) and seed-to-pulp ratio (r=0.73). Cultivar differences highlighted in the chronology of ripening events are critical for defining optimum harvest maturity and postharvest handling protocols for premium quality cactus pear fruit.

  15. The ability of a cold-adapted Rhodotorula mucilaginosa strain from Tibet to control blue mold in pear fruit.

    Science.gov (United States)

    Hu, Hao; Yan, Fujie; Wilson, Charles; Shen, Qing; Zheng, Xiaodong

    2015-12-01

    Cold-adapted yeasts were isolated from soil samples collected in Tibet and evaluated as potential biocontrol agents against blue mold (Penicillium expansum) of pear fruit in cold storage. YC1, an isolate identified as Rhodotorula mucilaginosa, was found to exhibit the greatest biocontrol activity among the different isolates that were screened. A washed cell suspension of YC1 exhibited the best biocontrol activity among three different preparations that were used in the current study. A concentration of 10(8) cells/ml reduced the incidence of decay to 35 %, compared to the control where decay incidence was 100 %. A higher intracellular level of trehalose and a higher proportion of polyunsaturated acids present in YC1, was associated with increased the tolerance of this strain to low temperatures, relative to the other strains that were evaluated. The increased tolerance to low temperature allowed the YC1 strain of yeast to more effectively compete for nutrients and space in wounded pear fruit that had been inoculated with spores of P. expansum and placed in cold storage. The present study demonstrated the ability to select cold-adapted yeasts from cold climates and use them as biocontrol agents of postharvest diseases of fruit placed in cold storage. PMID:26454432

  16. 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.

  17. Effectiveness of Azadirachtin (NeemAzal-T/S in Controlling Pear Psylla (Cacopsylla pyri and European Red Mite (Panonychus ulmi

    Directory of Open Access Journals (Sweden)

    Dejan Marčić

    2009-01-01

    Full Text Available Here we present the results of field trials conducted in Serbia to evaluate the effectiveness of a neem-based product, NeemAzal-T/S (containing azadirachtin-A as its active ingredient in the form of an emulsifiable concentrate against pear psylla (Cacopsylla pyri and European red mite (Panonychus ulmi. Efficacy evaluation against C. pyri was carried out in a commercial pear orchard of the Williams pear cultivar, located at Borkovac (Ruma. The insecticides were applied at BBCH 09 pear growth stage, several days before the beginning of hatching of the first generation larvae. The efficacy of azadirachtin was compared to that of mineral oil, abamectin and diflubenzuron. Efficacy evaluation 18 DAT showed total termination of egg laying by C. pyri after treatments with azadirachtin and abamectin, while some new (white eggs were found after treatment with mineral oil. Diflubenzuron treatment failed to fully stop egg laying, but the number of white eggs was significantly lower than it was in the control. Azadirachtin and abamectin achieved 100% efficacy, while the effectiveness of mineral oil was 97.4%, and that of diflubenzuron a mere 59%. All four insecticides significantly reduced the number of older (yellow eggs and larvae, the efficacy being 80.5-92.6% (yellow eggs, 69.8-79.3% (larvae I-III instar and 94.3-100% (larvae IV-V instar. In evaluation 38 DAT, azadirachtin,abamectin and mineral oil achieved 100% efficacy against white and yellow eggs, while diflubenzuron achieved 93% and 86.9% efficacy. All four insecticides were found to demonstrate high efficacy against I-III instar larvae (99.2-100%, but mineral oil treatmentalone achieved high efficacy against IV-V instar larvae (92.4% as well. Efficacy evaluation against P. ulmi was carried out in a commercial orchard of the Red Chief apple cultivar located at Morović (Šid. Azadirachtin efficacy in controlling a summerpopulation of European red mite was compared to a mineral oil, clofentezine and

  18. 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.

  19. 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. PMID:27566360

  20. A comparison of single molecule and amplification based sequencing of cancer transcriptomes.

    Directory of Open Access Journals (Sweden)

    Lee T Sam

    Full Text Available The second wave of next generation sequencing technologies, referred to as single-molecule sequencing (SMS, carries the promise of profiling samples directly without employing polymerase chain reaction steps used by amplification-based sequencing (AS methods. To examine the merits of both technologies, we examine mRNA sequencing results from single-molecule and amplification-based sequencing in a set of human cancer cell lines and tissues. We observe a characteristic coverage bias towards high abundance transcripts in amplification-based sequencing. A larger fraction of AS reads cover highly expressed genes, such as those associated with translational processes and housekeeping genes, resulting in relatively lower coverage of genes at low and mid-level abundance. In contrast, the coverage of high abundance transcripts plateaus off using SMS. Consequently, SMS is able to sequence lower- abundance transcripts more thoroughly, including some that are undetected by AS methods; however, these include many more mapping artifacts. A better understanding of the technical and analytical factors introducing platform specific biases in high throughput transcriptome sequencing applications will be critical in cross platform meta-analytic studies.

  1. 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.

  2. 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.

  3. 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. PMID:27027375

  4. 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.

  5. 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.

  6. 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.

  7. 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...

  8. 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.

  9. DEVELOPMENT OF THE STERILE INSECT TECHNIQUE TO MANAGE AN INVASIVE INSECT PEST, CACTOBLASTIS CACTORUM, ATTACKING PRICKLY PEAR CACTUS IN QUINTANA ROO, MEXICO, AND SOUTHEASTERN USA

    Science.gov (United States)

    The most successful classical biological control of weeds program has been the control of invasive prickly-pear cactus (Opuntia spp.) by the Argentine cactus moth Cactoblastis cactorum. However, the moth has now become an invasive pest in the southeastern USA and its ability to dramatically control ...

  10. Physical mapping of black spot disease resistance/susceptibility-related genome regions in Japanese pear (Pyrus pyrifolia) by BAC-FISH.

    Science.gov (United States)

    Yamamoto, Masashi; Terakami, Shingo; Takada, Norio; Yamamoto, Toshiya

    2016-06-01

    Black spot disease, caused by Alternaria alternata Japanese pear pathotype, is one of the most harmful diseases in Japanese pear cultivation. In the present study, the locations of black spot disease resistance/susceptibility-related genome regions were studied by fluorescence in situ hybridization using BAC clone (BAC-FISH) on Japanese pear (Pyrus pyrifolia (Burm. f.) Nakai) chromosomes. Root tips of self-pollinated seedlings of 'Osa Gold' were used as materials. Chromosome samples were prepared by the enzymatic maceration and air-drying method. The BAC clone adjacent to the black spot disease-related gene was labeled as a probe for FISH analysis. Black spot disease-related genome regions were detected in telomeric positions of two medium size chromosomes. These two sites and six telomeric 18S-5.8S-25S rDNA sites were located on different chromosomes as determined from the results of multi-color FISH. The effectiveness of the physical mapping of useful genes on pear chromosomes achieved by the BAC-FISH method was unequivocally demonstrated. PMID:27436955

  11. Internal browning in pear fruit (Pyrus communis L. cv Conference) may be a result of a limited availability of energy and antioxidants

    NARCIS (Netherlands)

    Veltman, R.H.; Lenthéric, I.; Plas, van der L.H.W.; Peppelenbos, H.W.

    2003-01-01

    Storage of pears (Pyrus communis) under hypoxia, especially in the presence of increased CO2 partial pressures, can lead to development of brown core. Disorder development, concentrations of ascorbic acid (AA) and adenosine triphosphate (ATP), and respiration were examined under various O-2 (0-21 kP

  12. Characterization of the nutritional components in fruit and cladode of selenium-enriched nutraceutical cactus pear fruit varieties grown on agricultural sediment

    Science.gov (United States)

    Different accessions of different colored cactus pear (Opuntia ficus Indica) were grown in soils high in salts, boron and selenium (Se) located in the Westside of central California. The changes in the nutritional status and biological transformation of the absorbed inorganic Se from the soils into ...

  13. Research on Fermentation Time in the Process of Pear Vinegar%梨醋工艺中发酵时间的研究

    Institute of Scientific and Technical Information of China (English)

    黄玉玲

    2014-01-01

    鸭梨醋是在发扬我国传统食醋文化的基础上研制成功的一种集鸭梨与醋保健功能为一体的新型保健饮料。主要研究以河南鸭梨为原料生产梨醋的工艺,通过实验确定了葡萄酒酵母进行酒精发酵的最佳时间为5天,确定了醋酸菌发酵时间为6天。%Pear vinegar is a new kind of healthy drink developed successfully with the healthy functions of pear and vinegar based on carrying forward Chinese traditional culture.This paper mainly studies the production process of pear vinegar,taking He′nan pears as raw materials.The optimum alcohol fermentation time of wine yeast is 5 days,and the acetic acid bacteria fermentation time is 6 days, which are determined by experiments.

  14. Selenium accumulation, distribution and speciation in spineless prickly pear cactus: a salt, boron, and drought tolerant, selenium-enriched nutraceutical fruit crop.

    Science.gov (United States)

    Prickly pear cactus (Opuntia) may be an alternative crop to grow in drainage-impacted regions of the westside of California, where high levels of salinity, selenium (Se), and boron (B) are present. Preliminary trials have demonstrated that Opuntia can tolerate the adverse soil conditions, while accu...

  15. In vitro colchicine-induced polyploid plantlet production and regeneration from leaf explants of the diploid pear (Pyrus communis L.) cultivar, 'Fertility'

    Science.gov (United States)

    Polyploid plantlets, including triploid, tetraploid and other polyploids, were induced from in vitro leaves of a European pear (Pyrus communis L.) cultivar 'Fertility' by a colchicine treatment. In vitro leaves were incubated in 0.4% colchicine solution for 24, 48 or 72 h, and transferred to an adv...

  16. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid diagnosis of chilli veinal mottle virus.

    Science.gov (United States)

    Banerjee, Amrita; Roy, Somnath; Sharma, Susheel Kumar; Dutta, Sudip Kumar; Chandra, Satish; Ngachan, S V

    2016-07-01

    Chilli veinal mottle virus (ChiVMV) causes significant economic loss to chilli cultivation in northeastern India, as well as in eastern Asia. In this study, we have developed a single-tube one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid, sensitive and specific diagnosis of ChiVMV. Amplification could be visualized after adding SYBR Green I (1000×) dye within 60 min under isothermal conditions at 63 °C, with a set of four primers designed based on the large nuclear inclusion protein (NIb) domain of ChiVMV (isolate KC-ML1). The RT-LAMP method was 100 times more sensitive than one-step reverse transcription polymerase chain reaction (RT-PCR), with a detection limit of 0.0001 ng of total RNA per reaction. PMID:27063408

  17. Low cost extraction and isothermal amplification of DNA for infectious diarrhea diagnosis.

    Science.gov (United States)

    Huang, Shichu; Do, Jaephil; Mahalanabis, Madhumita; Fan, Andy; Zhao, Lei; Jepeal, Lisa; Singh, Satish K; Klapperich, Catherine M

    2013-01-01

    In order to counter the common perception that molecular diagnostics are too complicated to work in low resource settings, we have performed a difficult sample preparation and DNA amplification protocol using instrumentation designed to be operated without wall or battery power. In this work we have combined a nearly electricity-free nucleic acid extraction process with an electricity-free isothermal amplification assay to detect the presence of Clostridium difficile (C. difficile) DNA in the stool of infected patients. We used helicase-dependent isothermal amplification (HDA) to amplify the DNA in a low-cost, thermoplastic reaction chip heated with a pair of commercially available toe warmers, while using a simple Styrofoam insulator. DNA was extracted from known positive and negative stool samples. The DNA extraction protocol utilized an air pressure driven solid phase extraction device run using a standard bicycle pump. The simple heater setup required no electricity or battery and was capable of maintaining the temperature at 65°C±2°C for 55 min, suitable for repeatable HDA amplification. Experiments were performed to explore the adaptability of the system for use in a range of ambient conditions. When compared to a traditional centrifuge extraction protocol and a laboratory thermocycler, this disposable, no power platform achieved approximately the same lower limit of detection (1.25×10(-2) pg of C. difficile DNA) while requiring much less raw material and a fraction of the lab infrastructure and cost. This proof of concept study could greatly impact the accessibility of molecular assays for applications in global health.

  18. Amplification and overexpression of JUNB is associated with primary cutaneous T-cell lymphomas.

    Science.gov (United States)

    Mao, Xin; Orchard, Guy; Lillington, Debra M; Russell-Jones, Robin; Young, Bryan D; Whittaker, Sean J

    2003-02-15

    Primary cutaneous lymphomas (PCLs) represent a heterogeneous group of extranodal T- and B-cell malignancies. The underlying molecular pathogenesis of this malignancy remains unclear. This study aimed to characterize oncogene abnormalities in PCLs. Using genomic microarray, we detected oncogene copy number gains of RAF1 (3p25), CTSB (8p22), PAK1 (11q13), and JUNB (19p13) in 5 of 7 cases of mycosis fungoides (MF)/Sezary syndrome (SS) (71%), gains of FGFR1 (8p11), PTPN (20q13), and BCR (22q11) in 4 cases (57%), and gains of MYCL1 (1p34), PIK3CA (3q26), HRAS (11p15), MYBL2 (20q13), and ZNF217 (20q13) in 3 cases (43%). Amplification of JUNB was studied in 104 DNA samples from 78 PCL cases using real-time polymerase chain reaction. Twenty-four percent of cases, including 7 of 10 cases of primary cutaneous CD30(+) anaplastic large-cell lymphoma (C-ALCL), 4 of 14 MF, 4 of 22 SS, and 2 of 23 primary cutaneous B-cell lymphoma (PCBCL) showed amplification of JUNB, and high-level amplification of this oncogene was present in 3 C-ALCL and 2 MF cases. JUNB protein expression was analyzed in tissue sections from 69 PCL cases, and 44% of cases, consisting of 21 of 23 SS, 6 of 8 C-ALCL, 5 of 10 MF, and 9 of 21 PCBCL, demonstrated nuclear expression of JUNB by tumor cells. Overexpression of JUNB also was detected in 5 C-ALCL and 2 SS cases. These results have revealed, for the first time, amplification and expression patterns of JUNB in PCL, suggesting that JUNB may be critical in the pathogenesis of primary cutaneous T-cell lymphomas.

  19. Low cost extraction and isothermal amplification of DNA for infectious diarrhea diagnosis.

    Directory of Open Access Journals (Sweden)

    Shichu Huang

    Full Text Available In order to counter the common perception that molecular diagnostics are too complicated to work in low resource settings, we have performed a difficult sample preparation and DNA amplification protocol using instrumentation designed to be operated without wall or battery power. In this work we have combined a nearly electricity-free nucleic acid extraction process with an electricity-free isothermal amplification assay to detect the presence of Clostridium difficile (C. difficile DNA in the stool of infected patients. We used helicase-dependent isothermal amplification (HDA to amplify the DNA in a low-cost, thermoplastic reaction chip heated with a pair of commercially available toe warmers, while using a simple Styrofoam insulator. DNA was extracted from known positive and negative stool samples. The DNA extraction protocol utilized an air pressure driven solid phase extraction device run using a standard bicycle pump. The simple heater setup required no electricity or battery and was capable of maintaining the temperature at 65°C±2°C for 55 min, suitable for repeatable HDA amplification. Experiments were performed to explore the adaptability of the system for use in a range of ambient conditions. When compared to a traditional centrifuge extraction protocol and a laboratory thermocycler, this disposable, no power platform achieved approximately the same lower limit of detection (1.25×10(-2 pg of C. difficile DNA while requiring much less raw material and a fraction of the lab infrastructure and cost. This proof of concept study could greatly impact the accessibility of molecular assays for applications in global health.

  20. Low cost extraction and isothermal amplification of DNA for infectious diarrhea diagnosis.

    Science.gov (United States)

    Huang, Shichu; Do, Jaephil; Mahalanabis, Madhumita; Fan, Andy; Zhao, Lei; Jepeal, Lisa; Singh, Satish K; Klapperich, Catherine M

    2013-01-01

    In order to counter the common perception that molecular diagnostics are too complicated to work in low resource settings, we have performed a difficult sample preparation and DNA amplification protocol using instrumentation designed to be operated without wall or battery power. In this work we have combined a nearly electricity-free nucleic acid extraction process with an electricity-free isothermal amplification assay to detect the presence of Clostridium difficile (C. difficile) DNA in the stool of infected patients. We used helicase-dependent isothermal amplification (HDA) to amplify the DNA in a low-cost, thermoplastic reaction chip heated with a pair of commercially available toe warmers, while using a simple Styrofoam insulator. DNA was extracted from known positive and negative stool samples. The DNA extraction protocol utilized an air pressure driven solid phase extraction device run using a standard bicycle pump. The simple heater setup required no electricity or battery and was capable of maintaining the temperature at 65°C±2°C for 55 min, suitable for repeatable HDA amplification. Experiments were performed to explore the adaptability of the system for use in a range of ambient conditions. When compared to a traditional centrifuge extraction protocol and a laboratory thermocycler, this disposable, no power platform achieved approximately the same lower limit of detection (1.25×10(-2) pg of C. difficile DNA) while requiring much less raw material and a fraction of the lab infrastructure and cost. This proof of concept study could greatly impact the accessibility of molecular assays for applications in global health. PMID:23555883

  1. Mtp-40 and alpha antigen gene fragment amplification for the detection of Mycobacterium tuberculosis in Colombian clinical specimens

    OpenAIRE

    Alfonso Rosalba; Romero Rosa Elena; Patarroyo Manuel Elkin; Murillo Luis Angel

    2002-01-01

    In this study, the use of Mtp-40 and alpha antigen polymerase chain reaction (PCR) amplification fragments for the precise tuberculosis (TB) diagnosis was evaluated. One hundred and ninety two different samples were obtained from 113 patients with suspected TB. Mtp-40 and alpha antigen protein genes were amplified by the PCR technique and compared to both the "gold standard" (culture) test, as well as the clinical parameters (including a clinical record and X-ray film exam in 113 patients). T...

  2. Diagnostic value of amplification of human cytomegalovirus DNA from gastrointestinal biopsies from human immunodeficiency virus-infected patients.

    OpenAIRE

    Cotte, L.; Drouet, E.; Bissuel, F.; Denoyel, G A; Trepo, C

    1993-01-01

    In order to assess the value of human cytomegalovirus (HCMV) DNA amplification of gastrointestinal biopsies, we studied 57 human immunodeficiency virus-infected patients with and without gastrointestinal HCMV diseases. After DNA extraction, a 406-bp fragment from the unique short region of the HCMV genome was amplified by 35 cycles of polymerase chain reaction (PCR) and semiquantified from 80 to 80,000 HCMV genomic copies. Among 12 non-AIDS patients, the PCR assay was negative for 11 of 12 du...

  3. Amplification and Asymmetry in Crashes and Frenzies

    OpenAIRE

    Han N. Ozsoylev

    2005-01-01

    We often observe disproportionate reactions to tangible information in large stock price movements. Moreover these movements feature an asymmetry: the number of crashes is more than that of frenzies in the S&P 500 index. This paper offers an explanation for these two characteristics of large movements in which hedging (portfolio insurance) causes amplified price reactions to news and liquidity shocks as well as an asymmetry biased towards crashes. Risk aversion of traders is shown to be essen...

  4. Different biosynthesis patterns among flavonoid 3-glycosides with distinct effects on accumulation of other flavonoid metabolites in pears (Pyrus bretschneideri Rehd..

    Directory of Open Access Journals (Sweden)

    Rui Zhai

    Full Text Available Flavonoid biosynthesis profile was clarified by fruit bagging and re-exposure treatments in the green Chinese pear 'Zaosu' (Pyrus bretschneideri Rehd. and its red mutant 'Red Zaosu'. Two distinct biosynthesis patterns of flavonoid 3-glycosides were found in 'Zaosu' pear. By comparison with 'Red Zaosu', the biosynthesis of flavonoid 3-galactosides and flavonoid 3-arabinosides were inhibited by bagging and these compounds only re-accumulated to a small degree in the fruit peel of 'Zaosu' after the bags were removed. In contrast, the biosynthesis of flavonoid 3-gluctosides and flavonoid 3-rutinosides was reduced by bagging and then increased when the fruits were re-exposed to sunlight. A combination of correlation, multicollinearity test and partial-correlation analyses among major flavonoid metabolites indicated that biosynthesis of each phenolic compound was independent in 'Zaosu' pear, except for the positive correlation between flavonoid 3-rutincosides and flavanols. In contrast with the green pear cultivar, almost all phenolic compounds in the red mutant had similar biosynthesis patterns except for arbutin. However, only the biosynthesis of flavonoid 3-galactosides was relatively independent and strongly affected the synthesis of the other phenolic compounds. Therefore, we propose a hypothesis that the strong accumulation of flavonoid 3-galactosides stimulated the biosynthesis of other flavonoid compounds in the red mutant and, therefore, caused systemic variation of flavonoid biosynthesis profiles between 'Zaosu' and its red mutant. This hypothesis had been further demonstrated by the enzyme activity of UFGT, and transcript levels of flavonoid biosynthetic genes and been well tested by a stepwise linear regression forecasting model. The gene that encodes flavonoid 3-galacosyltransferase was also identified and isolated from the pear genome.

  5. PyMYB10 and PyMYB10.1 Interact with bHLH to Enhance Anthocyanin Accumulation in Pears

    Science.gov (United States)

    Feng, Shouqian; Sun, Shasha; Chen, Xiaoliu; Wu, Shujing; Wang, Deyun; Chen, Xuesen

    2015-01-01

    Color is an important agronomic trait of pears, and the anthocyanin content of fruit is immensely significant for pear coloring. In this study, an anthocyanin-activating R2R3-MYB transcription factor gene, PyMYB10.1, was isolated from fruits of red sand pear (Pyrus pyrifolia cv. Aoguan). Alignments of the nucleotide and amino acid sequences suggested that PyMYB10.1 was involved in anthocyanin regulation. Similar to PyMYB10, PyMYB10.1 was predominantly expressed in red tissues, including the skin, leaf and flower, but it was minimally expressed in non-red fruit flesh. The expression of this gene could be induced by light. Dual-luciferase assays indicated that both PyMYB10 and PyMYB10.1 activated the AtDFR promoter. The activation of AtDFR increased to a greater extent when combined with a bHLH co-factor, such as PybHLH, MrbHLH1, MrbHLH2, or AtbHLH2. However, the response of this activation depended on the protein complex formed. PyMYB10-AtbHLH2 activated the AtDFR promoter to a greater extent than other combinations of proteins. PyMYB10-AtbHLH2 also induced the highest anthocyanin accumulation in tobacco transient-expression assays. Moreover, PybHLH interacted with PyMYB10 and PyMYB10.1. These results suggest that both PyMYB10 and PyMYB10.1 are positive anthocyanin biosynthesis regulators in pears that act via the formation of a ternary complex with PybHLH. The functional characterization of PyMYB10 and PyMYB10.1 will aid further understanding of the anthocyanin regulation in pears. PMID:26536358

  6. Study on the Processing Technology of Qinghai Soft Pear Brandy Wine%青海软儿梨白兰地果酒的工艺研究

    Institute of Scientific and Technical Information of China (English)

    孙万成; 罗毅皓

    2011-01-01

    [Objective] To produce Qinghai soft pear brandy wine. [ Method] The production technology of Qinghai soft pear brandy wine was optimized through a series of processes, including cleaning, juice squeezing, sugar and acid addition, fermentation, distillation, aging, filtration and so on. And then the effects of fermentation time, fermentation temperature and yeast inoculum on the quality of brandy wine were studied. [Result] The optimal conditions were determined through orthogonal experiment, which were 20 ℃ fermentation temperature, 10 d fermentation time and 0.3% yeast inoculum, the products produced under those conditions had the external features of brandy and the special flavor of soft pear. [ Conclusion] The soft pear brandy wine with typical fragrance of wine and pleasant aroma of soft pear could be produced.%[目的]研制青海软儿梨白兰地果酒.[方法]以青海软儿梨为原料,经过清洗、榨汁、调糖、调酸、发酵、蒸馏、陈酿、过滤等工艺优化软儿梨白兰地果酒工艺,并研究发酵时间、发酵温度和酵母菌接种量对白兰地品质的影响.[结果]通过正交试验确定了最适澄清务件:发酵温度20 ℃,发酵时间10 d,酵母菌接种量0.3%,产品具有白兰地的外观特征和软梨儿的特色香味.[结论]可以生产出酒香纯正、果香宜人的软儿梨白兰地.

  7. PyMYB10 and PyMYB10.1 Interact with bHLH to Enhance Anthocyanin Accumulation in Pears.

    Directory of Open Access Journals (Sweden)

    Shouqian Feng

    Full Text Available Color is an important agronomic trait of pears, and the anthocyanin content of fruit is immensely significant for pear coloring. In this study, an anthocyanin-activating R2R3-MYB transcription factor gene, PyMYB10.1, was isolated from fruits of red sand pear (Pyrus pyrifolia cv. Aoguan. Alignments of the nucleotide and amino acid sequences suggested that PyMYB10.1 was involved in anthocyanin regulation. Similar to PyMYB10, PyMYB10.1 was predominantly expressed in red tissues, including the skin, leaf and flower, but it was minimally expressed in non-red fruit flesh. The expression of this gene could be induced by light. Dual-luciferase assays indicated that both PyMYB10 and PyMYB10.1 activated the AtDFR promoter. The activation of AtDFR increased to a greater extent when combined with a bHLH co-factor, such as PybHLH, MrbHLH1, MrbHLH2, or AtbHLH2. However, the response of this activation depended on the protein complex formed. PyMYB10-AtbHLH2 activated the AtDFR promoter to a greater extent than other combinations of proteins. PyMYB10-AtbHLH2 also induced the highest anthocyanin accumulation in tobacco transient-expression assays. Moreover, PybHLH interacted with PyMYB10 and PyMYB10.1. These results suggest that both PyMYB10 and PyMYB10.1 are positive anthocyanin biosynthesis regulators in pears that act via the formation of a ternary complex with PybHLH. The functional characterization of PyMYB10 and PyMYB10.1 will aid further understanding of the anthocyanin regulation in pears.

  8. EMISSION-LINE GALAXIES FROM THE HUBBLE SPACE TELESCOPE PROBING EVOLUTION AND REIONIZATION SPECTROSCOPICALLY (PEARS) GRISM SURVEY. II. THE COMPLETE SAMPLE

    Energy Technology Data Exchange (ETDEWEB)

    Pirzkal, Nor; Rothberg, Barry; Ly, Chun; Grogin, Norman A.; Dahlen, Tomas; Noeske, Kai G.; Bellini, Andrea [Space Telescope Science Institute, 3700 San Martin Drive, Baltimore, MD 21210 (United States); Malhotra, Sangeeta; Rhoads, James E.; Cohen, Seth H.; Mechtley, Matthew; Windhorst, Rogier A. [School of Earth And Space Exploration, Arizona State University, Tempe, AZ 85287-1404 (United States); Meurer, Gerhardt R. [International Centre for Radio Astronomy Research, The University of Western Australia, 35 Stirling Highway, Crawley, WA 6009 (Australia); Walsh, Jeremy R. [European Southern Observatory, Karl-Schwarzschild-Strasse 2, D-85748 Garching (Germany); Hathi, Nimish P. [Carnegie Observatories, 813 Santa Barbara Street, Pasadena, CA 91101 (United States); Holwerda, Benne W. [ESA-ESTEC, Keplerlaan 1, 2200 AG, Noordwijk (Netherlands); Straughn, Amber N. [Astrophysics Science Division, Goddard Space Flight Center, Code 665, Greenbelt, MD 20771 (United States)

    2013-07-20

    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 board Hubble Space Telescope. 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 complemented by the spectroscopic results. Using the PEARS data, we are able to identify star-forming galaxies (SFGs) within the redshift volume 0 < z < 1.5. Star-forming regions in the PEARS survey are pinpointed independently of the host galaxy. This method allows us to detect the presence of multiple emission-line regions (ELRs) within a single galaxy. We identified a total of 1162 H{alpha}, [O III], and/or [O II] emission lines in the PEARS sample of 906 galaxies to a limiting flux of {approx}10{sup -18} erg s{sup -1} cm{sup -2}. The ELRs have also been compared to the properties of the host galaxy, including morphology, luminosity, and mass. From this analysis, we find three key results: (1) the computed line luminosities show evidence of a flattening in the luminosity function with increasing redshift; (2) the star-forming systems show evidence of complex morphologies with star formation occurring predominantly within one effective (half-light) radius. However, the morphologies show no correlation with host stellar mass. (3) Also, the number density of SFGs with M{sub *} {>=} 10{sup 9} M{sub Sun} decreases by an order of magnitude at z {<=} 0.5 relative to the number at 0.5 < z < 0.9, supporting the argument of galaxy downsizing.

  9. Mechanism of Gene Amplification via Yeast Autonomously Replicating Sequences

    Directory of Open Access Journals (Sweden)

    Shelly Sehgal

    2015-01-01

    Full Text Available The present investigation was aimed at understanding the molecular mechanism of gene amplification. Interplay of fragile sites in promoting gene amplification was also elucidated. The amplification promoting sequences were chosen from the Saccharomyces cerevisiae ARS, 5S rRNA regions of Plantago ovata and P. lagopus, proposed sites of replication pausing at Ste20 gene locus of S. cerevisiae, and the bend DNA sequences within fragile site FRA11A in humans. The gene amplification assays showed that plasmid bearing APS from yeast and human beings led to enhanced protein concentration as compared to the wild type. Both the in silico and in vitro analyses were pointed out at the strong bending potential of these APS. In addition, high mitotic stability and presence of TTTT repeats and SAR amongst these sequences encourage gene amplification. Phylogenetic analysis of S. cerevisiae ARS was also conducted. The combinatorial power of different aspects of APS analyzed in the present investigation was harnessed to reach a consensus about the factors which stimulate gene expression, in presence of these sequences. It was concluded that the mechanism of gene amplification was that AT rich tracts present in fragile sites of yeast serve as binding sites for MAR/SAR and DNA unwinding elements. The DNA protein interactions necessary for ORC activation are facilitated by DNA bending. These specific bindings at ORC promote repeated rounds of DNA replication leading to gene amplification.

  10. A mechanism of gene amplification driven by small DNA fragments.

    Directory of Open Access Journals (Sweden)

    Kuntal Mukherjee

    Full Text Available DNA amplification is a molecular process that increases the copy number of a chromosomal tract and often causes elevated expression of the amplified gene(s. Although gene amplification is frequently observed in cancer and other degenerative disorders, the molecular mechanisms involved in the process of DNA copy number increase remain largely unknown. We hypothesized that small DNA fragments could be the trigger of DNA amplification events. Following our findings that small fragments of DNA in the form of DNA oligonucleotides can be highly recombinogenic, we have developed a system in the yeast Saccharomyces cerevisiae to capture events of chromosomal DNA amplification initiated by small DNA fragments. Here we demonstrate that small DNAs can amplify a chromosomal region, generating either tandem duplications or acentric extrachromosomal DNA circles. Small fragment-driven DNA amplification (SFDA occurs with a frequency that increases with the length of homology between the small DNAs and the target chromosomal regions. SFDA events are triggered even by small single-stranded molecules with as little as 20-nt homology with the genomic target. A double-strand break (DSB external to the chromosomal amplicon region stimulates the amplification event up to a factor of 20 and favors formation of extrachromosomal circles. SFDA is dependent on Rad52 and Rad59, partially dependent on Rad1, Rad10, and Pol32, and independent of Rad51, suggesting a single-strand annealing mechanism. Our results reveal a novel molecular model for gene amplification, in which small DNA fragments drive DNA amplification and define the boundaries of the amplicon region. As DNA fragments are frequently found both inside cells and in the extracellular environment, such as the serum of patients with cancer or other degenerative disorders, we propose that SFDA may be a common mechanism for DNA amplification in cancer cells, as well as a more general cause of DNA copy number variation

  11. Signal Amplification in Field Effect-Based Sandwich Enzyme-Linked Immunosensing by Tuned Buffer Concentration with Ionic Strength Adjuster.

    Science.gov (United States)

    Kumar, Satyendra; Kumar, Narendra; Panda, Siddhartha

    2016-04-01

    Miniaturization of the sandwich enzyme-based immunosensor has several advantages but could result in lower signal strength due to lower enzyme loading. Hence, technologies for amplification of the signal are needed. Signal amplification in a field effect-based electrochemical immunosensor utilizing chip-based ELISA is presented in this work. First, the molarities of phosphate buffer saline (PBS) and concentrations of KCl as ionic strength adjuster were optimized to maximize the GOx glucose-based enzymatic reactions in a beaker for signal amplification measured by change in the voltage shift with an EIS device (using 20 μl of solution) and validated with a commercial pH meter (using 3 ml of solution). The PBS molarity of 100 μM with 25 mM KCl provided the maximum voltage shift. These optimized buffer conditions were further verified for GOx immobilized on silicon chips, and similar trends with decreased PBS molarity were obtained; however, the voltage shift values obtained on chip reaction were lower as compared to the reactions occurring in the beaker. The decreased voltage shift with immobilized enzyme on chip could be attributed to the increased Km (Michaelis-Menten constant) values in the immobilized GOx. Finally, a more than sixfold signal enhancement (from 8 to 47 mV) for the chip-based sandwich immunoassay was obtained by altering the PBS molarity from 10 to 100 μM with 25 mM KCl. PMID:26801818

  12. Clinical application of a ligation-independent pathway of multiplex ligation-dependent probe amplification for the determination of quinolone susceptibility of Streptococcus pneumoniae.

    Science.gov (United States)

    Uno, Naoki; Araki, Nobuko; Kaku, Norihito; Kosai, Kosuke; Hasegawa, Hiroo; Yanagihara, Katsunori

    2016-09-01

    We previously uncovered a ligation-independent pathway of multiplex ligation-dependent probe amplification (MLPA) through which products of MLPA could be amplified without both hybridization and ligation reactions. Here, we utilized this pathway to detect an antibiotic resistance mutation of quinolones in Streptococcus pneumoniae. PMID:27343683

  13. Complementary weak-value amplification with concatenated postselections

    CERN Document Server

    Viza, Gerardo I; Liu, Wei-Tao; Howell, John C

    2016-01-01

    We measure a transverse momentum kick in a Sagnac interferometer using weak-value amplification with two postselections. The first postselection is controlled by a polarization dependent phase mismatch between both paths of a Sagnac interferometer and the second postselection is controlled by a polarizer at the exit port. By monitoring the darkport of the interferometer, we study the complementary amplification of the concatenated postselections, where the polarization extinction ratio is greater than the contrast of the spatial interference. In this case, we find an improvement in the amplification of the signal of interest by introducing a second postselection to the system.

  14. Amplification of Spin Waves by Thermal Spin-Transfer Torque

    Science.gov (United States)

    Padrón-Hernández, E.; Azevedo, A.; Rezende, S. M.

    2011-11-01

    We observe amplification of spin-wave packets propagating along a film of single-crystal yttrium iron garnet subject to a transverse temperature gradient. The spin waves are excited and detected with standard techniques used in magnetostatic microwave delay lines in the 1-2 GHz frequency range. The amplification is attributed to the action of a thermal spin-transfer torque acting on the magnetization that opposes the relaxation and which is created by spin currents generated through the spin-Seebeck effect. The experimental data are interpreted with a spin-wave model that gives an amplification gain in very good agreement with the data.

  15. Amplification and chromosomal dispersion of human endogenous retroviral sequences

    International Nuclear Information System (INIS)

    Endogenous retroviral sequences have undergone amplification events involving both viral and flanking cellular sequences. The authors cloned members of an amplified family of full-length endogenous retroviral sequences. Genomic blotting, employing a flanking cellular DNA probe derived from a member of this family, revealed a similar array of reactive bands in both humans and chimpanzees, indicating that an amplification event involving retroviral and associated cellular DNA sequences occurred before the evolutionary separation of these two primates. Southern analyses of restricted somatic cell hybrid DNA preparations suggested that endogenous retroviral segments are widely dispersed in the human genome and that amplification and dispersion events may be linked

  16. PCR amplification on microarrays of gel immobilized oligonucleotides

    Science.gov (United States)

    Strizhkov, Boris; Tillib, Sergei; Mikhailovich, Vladimir; Mirzabekov, Andrei

    2003-11-04

    The invention relates two general methods for performing PCR amplification, combined with the detection and analysis of the PCR products on a microchip. In the first method, the amplification occurs both outside and within a plurality of gel pads on a microchip, with at least one oligonucleotide primer immobilized in a gel pad. In the second method, PCR amplification also takes place within gel pads on a microchip, but the pads are surrounded by a hydrophobic liquid such as that which separates the individual gel pads into environments which resemble micro-miniaturized test tubes.

  17. Protein enrichment of cactus pear (Opuntia ficus - indica Mill using Saccharomyces cerevisiae in solid-state fermentation

    Directory of Open Access Journals (Sweden)

    Lúcia de Fátima Araújo

    2005-06-01

    Full Text Available The microbial protein bioconversion of cactus pear by yeast in solid medium was studied. Three cultivation variables used were: inoculum's concentrations (5, 10 and 15 %, substrate layer thickness (2, 4 and 6 cm and temperature (30, 34 and 38 ºC. The rate of dry matter production and total protein were determined. Results obtained were variance analysis, gross energy and in vitro dry matter digestibility. The maximum protein amount achieved for the conditions studied in the present work was higher than 26 %, which was compatible or greater than those of conventional concentrates of protein supplements used for animal feed. The protein concentrate of cactus pear had a higher in vitro digestibility index (95.8 % and did not show any changes in the gross energy value when compared to that of the cactus pear in natura.A bioconversão da proteína microbiana através da levedura em meio sólido, foi estudada em palma forrageira cultivada em condições laboratoriais, sob três níveis de concentração do inóculo (5, 10 e 15%, espessuras distintas das camadas dos substratos (2, 4 e 6cm e temperaturas (30, 34 e 38ºC. Foram analisadas as taxas de produção de matéria seca (MS, proteína bruta (PB, cujos resultados foram submetidos à análise de variância, energia bruta (EB e digestibilidade in vitro da matéria seca (DIVMS. O valor máximo de teor protéico, alcançado nas condições estudadas nesse trabalho, foi superior a 26%, sendo esse teor compatível ou maior do que os concentrados convencionais utilizados como suplemento protéico para a ração animal. O concentrado protéico da palma obteve um alto índice de digestibilidade in vitro (95,8% e não apresentou grande alteração no valor da energia bruta se comparada com a palma in natura.

  18. Targeted deep resequencing identifies coding variants in the PEAR1 gene that play a role in platelet aggregation.

    Directory of Open Access Journals (Sweden)

    Yoonhee Kim

    Full Text Available Platelet aggregation is heritable, and genome-wide association studies have detected strong associations with a common intronic variant of the platelet endothelial aggregation receptor1 (PEAR1 gene both in African American and European American individuals. In this study, we used a sequencing approach to identify additional exonic variants in PEAR1 that may also determine variability in platelet aggregation in the GeneSTAR Study. A 0.3 Mb targeted region on chromosome 1q23.1 including the entire PEAR1 gene was Sanger sequenced in 104 subjects (45% male, 49% African American, age = 52±13 selected on the basis of hyper- and hypo- aggregation across three different agonists (collagen, epinephrine, and adenosine diphosphate. Single-variant and multi-variant burden tests for association were performed. Of the 235 variants identified through sequencing, 61 were novel, and three of these were missense variants. More rare variants (MAF<5% were noted in African Americans compared to European Americans (108 vs. 45. The common intronic GWAS-identified variant (rs12041331 demonstrated the most significant association signal in African Americans (p = 4.020×10(-4; no association was seen for additional exonic variants in this group. In contrast, multi-variant burden tests indicated that exonic variants play a more significant role in European Americans (p = 0.0099 for the collective coding variants compared to p = 0.0565 for intronic variant rs12041331. Imputation of the individual exonic variants in the rest of the GeneSTAR European American cohort (N = 1,965 supports the results noted in the sequenced discovery sample: p = 3.56×10(-4, 2.27×10(-7, 5.20×10(-5 for coding synonymous variant rs56260937 and collagen, epinephrine and adenosine diphosphate induced platelet aggregation, respectively. Sequencing approaches confirm that a common intronic variant has the strongest association with platelet aggregation in African Americans

  19. Pigmentation in sand pear (Pyrus pyrifolia) fruit: biochemical characterization, gene discovery and expression analysis with exocarp pigmentation mutant.

    Science.gov (United States)

    Wang, Yue-zhi; Zhang, Shujun; Dai, Mei-song; Shi, Ze-bin

    2014-05-01

    Exocarp color of sand pear is an important trait for the fruit production and has caused our concern for a long time. Our previous study explored the different expression genes between the two genotypes contrasting for exocarp color, which indicated the different suberin, cutin, wax and lignin biosynthesis between the russet- and green-exocarp. In this study, we carried out microscopic observation and Fourier transform infrared spectroscopy analysis to detect the differences of tissue structure and biochemical composition between the russet- and green-exocarp of sand pear. The green exocarp was covered with epidermis and cuticle which was replaced by a cork layer on the surface of russet exocarp, and the chemicals of the russet exocarp were characterized by lignin, cellulose and hemicellulose. We explored differential gene expression between the russet exocarp of 'Niitaka' and its green exocarp mutant cv. 'Suisho' using Illumina RNA-sequencing. A total of 559 unigenes showed different expression between the two types of exocarp, and 123 of them were common to the previous study. The quantitative real time-PCR analysis supports the RNA-seq-derived gene with different expression between the two types of exocarp and revealed the preferential expression of these genes in exocarp than in mesocarp and fruit core. Gene ontology enrichment analysis revealed divorced expression of lipid metabolic process genes, transport genes, stress responsive genes and other biological process genes in the two types of exocarp. Expression changes in lignin metabolism-related genes were consistent with the different pigmentation of russet and green exocarp. Increased transcripts of putative genes involved the suberin, cutin and wax biosynthesis in 'Suisho' exocarp could facilitate deposition of the chemicals and take a role in the mutant trait responsible for the green exocarp. In addition, the divorced expression of ATP-binding cassette transporters involved in the trans

  20. Thermal amplification of field-correlation harvesting

    CERN Document Server

    Brown, Eric G

    2013-01-01

    We study the harvesting of quantum and classical correlations from a hot scalar field in a periodic cavity by a pair of spatially separated oscillator-detectors. Specifically, we utilize non-perturbative and exact (non-numerical) techniques to solve for the evolution of the detectors-field system and then we examine how the entanglement, Gaussian quantum discord, and mutual information obtained by the detectors change with the temperature of the field. While (as expected) the harvested entanglement rapidly decays to zero as temperature is increased, we find remarkably that both the mutual information and the discord can actually be increased by multiple orders of magnitude via increasing the temperature. We go on to explain this phenomenon by taking advantage of the translational invariance of the field and use this to make accurate predictions of the behavior of thermal amplification; by this we also introduce a new perspective on field-correlation harvesting that we feel is worthy of consideration in its ow...