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Sample records for one-step reverse transcription

  1. Selective control of primer usage in multiplex one-step reverse transcription PCR

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    Paul Natasha

    2009-12-01

    Full Text Available Abstract Background Multiplex RT-PCR is a valuable technique used for pathogen identification, disease detection and relative quantification of gene expression. The simplification of this protocol into a one-step procedure saves time and reagents. However, intensive PCR optimization is often required to overcome competing undesired PCR primer extension during the RT step. Results Herein, we report multiplex one-step RT-PCR experiments in which the PCR primers contain thermolabile phosphotriester modification groups. The presence of these groups minimizes PCR primer extension during the RT step and allows for control of PCR primer extension until the more stringent, elevated temperatures of PCR are reached. Results reveal that the use of primers whose extension can be controlled in a temperature-mediated way provides improved one-step RT-PCR specificity in both singleplex and multiplex reaction formats. Conclusions The need for an accurate and sensitive technique to quantify mRNA expression levels makes the described modified primer technology a promising tool for use in multiplex one-step RT-PCR. A more accurate representation of the abundances in initial template sample is feasible with modified primers, as artifacts of biased PCR are reduced because of greater improvements in reaction specificity.

  2. Rapid detection of sacbrood virus (SBV by one-step reverse transcription loop-mediated isothermal amplification assay

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    Jin-Long Yang

    2012-02-01

    Full Text Available Abstract Background Sacbrood virus (SBV primarily infects honeybee broods, and in order to deal with the problem cost effective detection methods are required. Findings A one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP assay was developed for the rapid identification of SBV. The data demonstrated that, in a simple water bath, SBV RNA could be detected as early as 20 min at 65°C, and a positive amplification reaction was visible to the naked eye due to a color change brought on by the addition of nucleic acid stain SYBR Green. Conclusions The current study presents a method for the rapid and simple detection of SBV by RT-LAMP with high sensitivity and analytic specificity.

  3. One-step reverse transcription loop-mediated isothermal amplification for the rapid detection of cucumber green mottle mosaic virus.

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    Li, Jin-yu; Wei, Qi-wei; Liu, Yong; Tan, Xin-qiu; Zhang, Wen-na; Wu, Jian-yan; Charimbu, Miriam Karwitha; Hu, Bai-shi; Cheng, Zhao-bang; Yu, Cui; Tao, Xiao-rong

    2013-11-01

    Cucumber green mottle mosaic virus (CGMMV) has caused serious damage to Cucurbitaceae crops worldwide. The virus is considered one of the most serious Cucurbitaceae quarantine causes in many countries. In this study, a highly efficient and practical one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed for the detection of CGMMV. The total RNA or crude RNA extracted from watermelon plants or seeds could be detected easily by this RT-LAMP assay. The RT-LAMP assay was conducted in isothermal (63°C) conditions within 1h. The amplified products of CGMMV could be detected as ladder-like bands using agarose gel electrophoresis or visualized in-tube under UV light with the addition of a fluorescent dye. The RT-LAMP amplification was specific to CGMMV, as no cross-reaction was observed with other viruses. The RT-LAMP assay was 100-fold more sensitive than that of reverse-transcription polymerase chain reaction (RT-PCR). This is the first report of the application of the RT-LAMP assay to detect CGMMV. The sensitive, specific and rapid RT-LAMP assay developed in this study can be applied widely in laboratories, the field and quarantine surveillance of CGMMV.

  4. Detection of Citrus leprosis virus C using specific primers and TaqMan probe in one-step real-time reverse-transcription polymerase chain reaction assays.

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    Choudhary, Nandlal; Wei, G; Govindarajulu, A; Roy, Avijit; Li, Wenbin; Picton, Deric D; Nakhla, M K; Levy, L; Brlansky, R H

    2015-11-01

    Citrus leprosis virus C (CiLV-C), a causal agent of the leprosis disease in citrus, is mostly present in the South and Central America and spreading toward the North America. To enable better diagnosis and inhibit the further spread of this re-emerging virus a quantitative (q) real-time reverse transcription polymerase chain reaction (qRT-PCR) assay is needed for early detection of CiLV-C when the virus is present in low titer in citrus leprosis samples. Using the genomic sequence of CiLV-C, specific primers and probe were designed and synthesized to amplify a 73 nt amplicon from the movement protein (MP) gene. A standard curve of the 73 nt amplicon MP gene was developed using known 10(10)-10(1) copies of in vitro synthesized RNA transcript to estimate the copy number of RNA transcript in the citrus leprosis samples. The one-step qRT-PCR detection assays for CiLV-C were determined to be 1000 times more sensitive when compared to the one-step conventional reverse transcription polymerase chain reaction (RT-PCR) CiLV-C detection method. To evaluate the quality of the total RNA extracts, NADH dehydrogenase gene specific primers (nad5) and probe were included in reactions as an internal control. The one-step qRT-PCR specificity was successfully validated by testing for the presence of CiLV-C in the total RNA extracts of the citrus leprosis samples collected from Belize, Costa Rica, Mexico and Panama. Implementation of the one-step qRT-PCR assays for CiLV-C diagnosis should assist regulatory agencies in surveillance activities to monitor the distribution pattern of CiLV-C in countries where it is present and to prevent further dissemination into citrus growing countries where there is no report of CiLV-C presence.

  5. A one-step reverse transcription loop-mediated isothermal amplification for detection and discrimination of infectious bursal disease virus

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    Qi Xiaole

    2011-03-01

    Full Text Available Abstract Background Infectious bursal disease (IBD is a highly contagious immunosuppressive disease in young chickens caused by infectious bursal disease virus (IBDV. It causes huge economic losses to the poultry industry. The objective of this study is to develop a loop-mediated isothermal amplification (LAMP method for the detection and discrimination of IBDV. Results In this study, we applied reverse transcription loop-mediated isothermal amplification (RT-LAMP to detect IBDV in one simple step and further identified the very virulent strain from non-vvIBDVs with a simply post-amplification restriction enzyme analysis. Based on sequence analysis, a set of two inner, two outer and two loop primers were designed to target the VP5 gene and they showed great specificity with no cross reaction to the other common avian pathogens. The detection limit determined by both color change inspection and agarose gel electrophoresis was 28 copies viral RNA, which was almost as sensitive as a real-time RT-PCR previous developed in our laboratory. We also identified a unique Tfi I restriction site located exclusively in non-vvIBDVs, so very virulent strain could be distinguished from current vaccine strains. By screening a panel of clinical specimens, results showed that this method is high feasible in clinical settings, and it obtained results 100% correlated with real-time RT-PCR. Conclusion RT-LAMP is a rapid, simple and sensitive assay. In combination with the Tfi I restriction analysis, this method holds great promises not only in laboratory detection and discrimination of IBDV but also in large scale field and clinical studies.

  6. A diagnostic one-step real-time reverse transcription polymerase chain reaction method for accurate detection of influenza virus type A

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    Behzadi, Mohammad Amin; Alborzi, Abdolvahab

    2016-01-01

    Introduction Influenza A is known as a public health concern worldwide. In this study, a novel one-step real-time reverse transcription polymerase chain reaction (rtRT-PCR) assay was designed and optimized for the detection of influenza A viruses. Material and methods The primers and probe were designed based on the analysis of 90 matrix nucleotide sequence data of influenza type A subtypes from the GenBank database of the National Center for Biotechnology Information (NCBI). The influenza virus A/Tehran/5652/2010 (H1N1 pdm09) was used as a reference. The rtRT-PCR assay was optimized, compared with that of the World Health Organization (WHO), and its analytical sensitivity, specificity and reproducibility were evaluated. In total, 64 nasopharyngeal swabs from patients with influenza-like illness (ILI) and 41 samples without ILI symptoms were tested for the virus, using conventional cell culture, direct immunofluorescence antibody (DFA) methods, and one-step rtRT-PCR with the designed primer set and probe and the WHO’s. Results The optimized assay results were similar to the WHO’s. The optimized assay results were similar to WHO’s, with non-significant differences for 10–103 copies of viral RNA/reaction (p > 0.05). It detected 10 copies of viral RNA/reaction with high reproducibility and no cross reactivity with other respiratory viruses. A specific cytopathic effect was observed in 6/64 (9.37%) of the ILI group using conventional culture and DFA staining methods; however, it was not seen in non-ILI. Also, the results of our assay and the WHO’s were similar to those of viral isolation and DFA staining. Conclusions Given the high specificity, sensitivity and reproducibility of this novel assay, it can serve as a reliable diagnostic tool for the detection of influenza A viruses in clinical specimens and lab experiments. PMID:27904520

  7. Establishment of a novel one-step reverse transcription loop-mediated isothermal amplification assay for rapid identification of RNA from the severe fever with thrombocytopenia syndrome virus.

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    Xu, Haihong; Zhang, Lei; Shen, Guangqiang; Feng, Cen; Wang, Xinying; Yan, Jie; Zhang, Yanjun

    2013-12-01

    As an emerging infectious disease, severe fever with thrombocytopenia syndrome virus (SFTSV) infection has been found in many areas of China. Suitable laboratory diagnostic method is urgently needed in clinical detections and epidemiological investigations. In this study, a modified, low-cost and rapid visualized one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for the detection of RNA from the SFTSV has been established. In order to avoid the risk of aerosol contamination and facilitate the naked eye to observe, a microcrystalline wax-dye capsule wrapping the highly sensitive DNA fluorescence dye SYBR Green I was added to the RT-LAMP reaction tube before the initiation of the assay. The detection limit of the established RT-LAMP assay was 10 fg template RNA per reaction mixture. The RT-LAMP assay was confirmed to be high specific to SFTSV, and no cross-reaction was found with the detection of the Chikungunya fever virus, Hemorrhagic Fever with Renal Syndrome virus (HFRSV), and Dengue fever virus. The assay was then applied for the detection of SFTSV RNA in 32 clinical serum samples and showed 94.4% consistence with the detection results of the real-time RT-PCR. The whole process, from sample preparation to result reporting, can be completed within 2h. This adapted, cost efficient and quick visualized RT-LAMP method is feasible for SFTSV field diagnosis in resource-limited field settings.

  8. Visual Detection of Potato leafroll virus by One-step Reverse Transcription Loop-Mediated Isothermal Amplification of DNA with Hydroxynaphthol Blue Dye

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    Ahmadi, S.; Almasi, A.M.; Fatehi, F.; Struik, P.C.; Moradi, A.

    2013-01-01

    Loop-mediated isothermal amplification (LAMP) assay is a novel technique for amplifying DNA under constant temperature, with high specificity, sensitivity, rapidity and efficiency. We applied reverse transcription loop-mediated isothermal amplification (RT-LAMP) to visually detect Potato leafroll vi

  9. Rapid detection and quantification of Ebola Zaire virus by one-step real-time quantitative reverse transcription-polymerase chain reaction.

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    Ro, Young-Tae; Ticer, Anysha; Carrion, Ricardo; Patterson, Jean L

    2017-04-01

    Given that Ebola virus causes severe hemorrhagic fever in humans with mortality rates as high as 90%, rapid and accurate detection of this virus is essential both for controlling infection and preventing further transmission. Here, a one-step qRT-PCR assay for rapid and quantitative detection of an Ebola Zaire strain using GP, VP24 or VP40 genes as a target is introduced. Routine assay conditions for hydrolysis probe detection were established from the manufacturer's protocol used in the assays. The analytical specificity and sensitivity of each assay was evaluated using in vitro synthesized viral RNA transcripts. The assays were highly specific for the RNA transcripts, no cross-reactivity being observed among them. The limits of detection of the assays ranged from 10(2) to 10(3) copies per reaction. The assays were also evaluated using viral RNAs extracted from cell culture-propagated viruses (Ebola Zaire, Sudan and Reston strains), confirming that they are gene- and strain-specific. The RT-PCR assays detected viral RNAs in blood samples from virus-infected animal, suggesting that they can be also a useful method for identifying Ebola virus in clinical samples. © 2017 The Societies and John Wiley & Sons Australia, Ltd.

  10. One-step detection of Bean pod mottle virus in soybean seeds by the reverse-transcription loop-mediated isothermal amplification

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    Wei Qi-Wei

    2012-09-01

    Full Text Available Abstract Background Bean pod mottle virus (BPMV is a wide-spread and destructive virus that causes huge economic losses in many countries every year. A sensitive, reliable and specific method for rapid surveillance is urgently needed to prevent further spread of BPMV. Methods A degenerate reverse-transcription loop-mediated isothermal amplification (RT-LAMP primer set was designed on the conserved region of BPMV CP gene. The reaction conditions of RT-LAMP were optimized and the feasibility, specificity and sensitivity of this method to detect BPMV were evaluated using the crude RNA rapidly extracted from soybean seeds. Results The optimized RT-LAMP parameters including 6 mM MgCl2, 0.8 M betaine and temperature at 62.5-65°C could successfully amplify the ladder-like bands from BPMV infected soybean seeds. The amplification was very specific to BPMV that no cross-reaction was observed with other soybean viruses. Inclusion of a fluorescent dye makes it easily be detected in-tube by naked eye. The sensitivity of RT-LAMP assay is higher than the conventional RT-PCR under the conditions tested, and the conventional RT-PCR couldn’t be used for detection of BPMV using crude RNA extract from soybean seeds. Conclusion A highly efficient and practical method was developed for the detection of BPMV in soybean seeds by the combination of rapid RNA extraction and RT-LAMP. This RT-LAMP method has great potential for rapid BPMV surveillance and will assist in preventing further spread of this devastating virus.

  11. A broad spectrum, one-step reverse-transcription PCR amplification of the neuraminidase gene from multiple subtypes of influenza A virus

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    Chen Wenbin

    2008-07-01

    Full Text Available Abstract Background The emergence of high pathogenicity strains of Influenza A virus in a variety of human and animal hosts, with wide geographic distribution, has highlighted the importance of rapid identification and subtyping of the virus for outbreak management and treatment. Type A virus can be classified into subtypes according to the viral envelope glycoproteins, hemagglutinin and neuraminidase. Here we review the existing specificity and amplification of published primers to subtype neuraminidase genes and describe a new broad spectrum primer pair that can detect all 9 neuraminidase subtypes. Results Bioinformatic analysis of 3,337 full-length influenza A neuraminidase segments in the NCBI database revealed semi-conserved regions not previously targeted by primers. Two degenerate primers with M13 tags, NA8F-M13 and NA10R-M13 were designed from these regions and used to generate a 253 bp cDNA product. One-step RT-PCR testing was successful in 31/32 (97% cases using a touchdown protocol with RNA from over 32 different cultured influenza A virus strains representing the 9 neuraminidase subtypes. Frozen blinded clinical nasopharyngeal aspirates were also assayed and were mostly of subtype N2. The region amplified was direct sequenced and then used in database searches to confirm the identity of the template RNA. The RT-PCR fragment generated includes one of the mutation sites related to oseltamivir resistance, H274Y. Conclusion Our one-step RT-PCR assay followed by sequencing is a rapid, accurate, and specific method for detection and subtyping of different neuraminidase subtypes from a range of host species and from different geographical locations.

  12. Microchip capillary electrophoresis with laser-induced fluorescence combined with one-step duplex reverse-transcription polymerase chain reaction for the rapid detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens.

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    Jia, Ruan; Chengjun, Sun; Heng, Chen; Chen, Zhou; Yuanqian, Li; Yongxin, Li

    2015-07-01

    Enterovirus 71 and Coxsackievirus A16 are the main pathogens causing hand-foot-mouth disease. In this paper, microchip capillary electrophoresis with laser-induced fluorescence combined with one-step duplex reverse transcript-polymerase chain reaction has been developed for the detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens. The specific reverse transcription-polymerase chain reaction amplicons labeled with SYBR Orange were separated by microchip capillary electrophoresis and detected by laser induced fluorescence detector within 7 min. The intraday and interday relative standard deviation of migration time for DNA Marker was in the range of 1.36-2.94 and 2.78-3.96%, respectively. The detection limits were as low as 2.06 × 10(3) copies/mL for Enterovirus 71 and 5 × 10(3) copies/mL for Coxsackievirus A16. No cross-reactivity was observed with rotavirus, astrovirus, norovirus, and adenovirus, which showed good specificity of the method. This assay was validated using 100 throat swab specimens that were detected by real-time reverse-transcript polymerase chain reaction in parallel and the two methods produced the same results. This study provided a rapid, sensitive and specific method for the detection of Enterovirus 71 and Coxsackievirus A16, which make a contribution to significant time and cost saving for the identification and treatment of patients.

  13. Evaluation and application of a one-step duplex real-time reverse transcription polymerase chain reaction assay for the rapid detection of influenza A (H7N9) virus from poultry samples.

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    Bao, Hongmei; Ma, Yong; Shi, Jianzhong; Zeng, Xianying; Zhao, Yuhui; Wang, Xiurong; Chen, Hualan

    2015-10-01

    In China, a novel reassortant influenza A (H7N9) virus, which has caused 435 cases of human infection, has recently emerged. Most cases of human infections with the H7N9 virus are known to be associated with a poultry farm and live-poultry markets. In this study, a one-step duplex real-time reverse transcription polymerase chain reaction (RRT-PCR) assay was developed for the simultaneous detection of the hemagglutinin (HA) and neuraminidase (NA) genes of the H7N9 virus for effective surveillance and early diagnosis of cases from clinical samples collected from live-poultry markets or poultry farms. The detection limit of this assay was as low as 0.1 EID50 of H7N9 viruses, which is similar to the detection limit of the real-time RT-PCR assay released by the Word Health Organization. The coefficients of variation (CVs) of both inter-assay and intra-assay reproducibility were less than 1.55 %, showing good reproducibility. No cross-reactivity was observed with RNA of other subtypes of influenza virus or other avian respiratory viruses. The assay can effectively detect H7N9 influenza virus RNA from multiple sources, including chickens, pigeons, ducks, humans, and the environment. Furthermore, the RRT-PCR assay was evaluated with more than 700 clinical samples collected from live-poultry markets and 120 experimentally infected chicken samples. Together, these results indicate that the duplex RRT-PCR assay is a specific, sensitive, and efficient diagnostic method for the epidemiological surveillance and diagnosis of H7N9 virus from different sources, particularly poultry samples.

  14. The use of a one-step real-time reverse transcription polymerase chain reaction (rRT-PCR) for the surveillance of viral hemorrhagic septicemia virus (VHSV) in Minnesota.

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    Phelps, Nicholas B D; Patnayak, Devi P; Jiang, Yin; Goyal, Sagar M

    2012-12-01

    Viral hemorrhagic septicemia virus (VHSV) is a highly contagious and pathogenic virus of fish. The virus infects more than 70 fish species worldwide, in both fresh and salt water. A new viral strain (VHSV-IVb) has proven both virulent and persistent, spreading throughout the Great Lakes of North America and to inland water bodies in the region. To better understand the geographic distribution of the virus, we used a modified real-time reverse transcription polymerase chain reaction (rRT-PCR) assay for high-throughput testing of fish for VHSV. The assay was shown to be twice as sensitive as the gold standard, virus isolation, and did not cross react with other viruses found in fish. In addition, the diagnostic turnaround time was reduced from 28 to 30 d for virus isolation to 2-4 d for rRT-PCR. To demonstrate the usefulness of the rRT-PCR assay, 115 high-priority water bodies in Minnesota were tested by both methods from April 2010 to June 2011. All survey sites tested negative for VHSV by both methods. The survey results have informed fisheries managers on the absence of VHSV in Minnesota and have better prepared them for the eventual arrival of the disease. In addition, the results demonstrate the value of this rRT-PCR as a surveillance tool to rapidly identify an outbreak so that it can be controlled in a timely manner.

  15. Development and evaluation of a real-time one step Reverse-Transcriptase PCR for quantitation of Chandipura Virus

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    Tandale Babasaheb V

    2008-12-01

    Full Text Available Abstract Background Chandipura virus (CHPV, a member of family Rhabdoviridae was attributed to an explosive outbreak of acute encephalitis in children in Andhra Pradesh, India in 2003 and a small outbreak among tribal children from Gujarat, Western India in 2004. The case-fatality rate ranged from 55–75%. Considering the rapid progression of the disease and high mortality, a highly sensitive method for quantifying CHPV RNA by real-time one step reverse transcriptase PCR (real-time one step RT-PCR using TaqMan technology was developed for rapid diagnosis. Methods Primers and probe for P gene were designed and used to standardize real-time one step RT-PCR assay for CHPV RNA quantitation. Standard RNA was prepared by PCR amplification, TA cloning and run off transcription. The optimized real-time one step RT-PCR assay was compared with the diagnostic nested RT-PCR and different virus isolation systems [in vivo (mice in ovo (eggs, in vitro (Vero E6, PS, RD and Sand fly cell line] for the detection of CHPV. Sensitivity and specificity of real-time one step RT-PCR assay was evaluated with diagnostic nested RT-PCR, which is considered as a gold standard. Results Real-time one step RT-PCR was optimized using in vitro transcribed (IVT RNA. Standard curve showed linear relationship for wide range of 102-1010 (r2 = 0.99 with maximum Coefficient of variation (CV = 5.91% for IVT RNA. The newly developed real-time RT-PCR was at par with nested RT-PCR in sensitivity and superior to cell lines and other living systems (embryonated eggs and infant mice used for the isolation of the virus. Detection limit of real-time one step RT-PCR and nested RT-PCR was found to be 1.2 × 100 PFU/ml. RD cells, sand fly cells, infant mice, and embryonated eggs showed almost equal sensitivity (1.2 × 102 PFU/ml. Vero and PS cell-lines (1.2 × 103 PFU/ml were least sensitive to CHPV infection. Specificity of the assay was found to be 100% when RNA from other viruses or healthy

  16. Chitosan-coated magnetic nanoparticles prepared in one step by reverse microemulsion precipitation.

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    López, Raúl G; Pineda, María G; Hurtado, Gilberto; León, Ramón Díaz de; Fernández, Salvador; Saade, Hened; Bueno, Darío

    2013-09-27

    Chitosan-coated magnetic nanoparticles (CMNP) were obtained at 70 °C and 80 °C in a one-step method, which comprises precipitation in reverse microemulsion in the presence of low chitosan concentration in the aqueous phase. X-ray diffractometry showed that CMNP obtained at both temperatures contain a mixture of magnetite and maghemite nanoparticles with ≈4.5 nm in average diameter, determined by electron microscopy, which suggests that precipitation temperature does not affect the particle size. The chitosan coating on nanoparticles was inferred from Fourier transform infrared spectrometry measurements; furthermore, the carbon concentration in the nanoparticles allowed an estimation of chitosan content in CMNP of 6%-7%. CMNP exhibit a superparamagnetic behavior with relatively high final magnetization values (≈49-53 emu/g) at 20 kOe and room temperature, probably due to a higher magnetite content in the mixture of magnetic nanoparticles. In addition, a slight direct effect of precipitation temperature on magnetization was identified, which was ascribed to a possible higher degree of nanoparticles crystallinity as temperature at which they are obtained increases. Tested for Pb2+ removal from a Pb(NO3)2 aqueous solution, CMNP showed a recovery efficacy of 100%, which makes them attractive for using in heavy metals ion removal from waste water.

  17. tCRISPRi: tunable and reversible, one-step control of gene expression

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    Li, Xin-Tian; Jun, Yonggun; Erickstad, Michael J.; Brown, Steven D.; Parks, Adam; Court, Donald L.; Jun, Suckjoon

    2016-12-01

    The ability to control the level of gene expression is a major quest in biology. A widely used approach employs deletion of a nonessential gene of interest (knockout), or multi-step recombineering to move a gene of interest under a repressible promoter (knockdown). However, these genetic methods are laborious, and limited for quantitative study. Here, we report a tunable CRISPR-cas system, “tCRISPRi”, for precise and continuous titration of gene expression by more than 30-fold. Our tCRISPRi system employs various previous advancements into a single strain: (1) We constructed a new strain containing a tunable arabinose operon promoter PBAD to quantitatively control the expression of CRISPR-(d)Cas protein over two orders of magnitude in a plasmid-free system. (2) tCRISPRi is reversible, and gene expression is repressed under knockdown conditions. (3) tCRISPRi shows significantly less than 10% leaky expression. (4) Most important from a practical perspective, construction of tCRISPRi to target a new gene requires only one-step of oligo recombineering. Our results show that tCRISPRi, in combination with recombineering, provides a simple and easy-to-implement tool for gene expression control, and is ideally suited for construction of both individual strains and high-throughput tunable knockdown libraries.

  18. A One-Step System for Convenient and Flexible Assembly of Transcription Activator-Like Effector Nucleases (TALENs).

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    Zhao, Jinlong; Sun, Wenye; Liang, Jing; Jiang, Jing; Wu, Zhao

    2016-09-01

    Transcription activator-like effector nucleases (TALENs) are powerful tools for targeted genome editing in diverse cell types and organisms. However, the highly identical TALE repeat sequences make it challenging to assemble TALEs using conventional cloning approaches, and multiple repeats in one plasmid are easily catalyzed for homologous recombination in bacteria. Although the methods for TALE assembly are constantly improving, these methods are not convenient because of laborious assembly steps or large module libraries, limiting their broad utility. To overcome the barrier of multiple assembly steps, we report a one-step system for the convenient and flexible assembly of a 180 TALE module library. This study is the first demonstration to ligate 9 mono-/dimer modules and one circular TALEN backbone vector in a one step process, generating 9.5 to 18.5 repeat sequences with an overall assembly rate higher than 50%. This system makes TALEN assembly much simpler than the conventional cloning of two DNA fragments because this strategy combines digestion and ligation into one step using circular vectors and different modules to avoid gel extraction. Therefore, this system provides a convenient tool for the application of TALEN-mediated genome editing in scientific studies and clinical trials.

  19. FusX: A Rapid One-Step Transcription Activator-Like Effector Assembly System for Genome Science.

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    Ma, Alvin C; McNulty, Melissa S; Poshusta, Tanya L; Campbell, Jarryd M; Martínez-Gálvez, Gabriel; Argue, David P; Lee, Han B; Urban, Mark D; Bullard, Cassandra E; Blackburn, Patrick R; Man, Toni K; Clark, Karl J; Ekker, Stephen C

    2016-06-01

    Transcription activator-like effectors (TALEs) are extremely effective, single-molecule DNA-targeting molecular cursors used for locus-specific genome science applications, including high-precision molecular medicine and other genome engineering applications. TALEs are used in genome engineering for locus-specific DNA editing and imaging, as artificial transcriptional activators and repressors, and for targeted epigenetic modification. TALEs as nucleases (TALENs) are effective editing tools and offer high binding specificity and fewer sequence constraints toward the targeted genome than other custom nuclease systems. One bottleneck of broader TALE use is reagent accessibility. For example, one commonly deployed method uses a multitube, 5-day assembly protocol. Here we describe FusX, a streamlined Golden Gate TALE assembly system that (1) is backward compatible with popular TALE backbones, (2) is functionalized as a single-tube 3-day TALE assembly process, (3) requires only commonly used basic molecular biology reagents, and (4) is cost-effective. More than 100 TALEN pairs have been successfully assembled using FusX, and 27 pairs were quantitatively tested in zebrafish, with each showing high somatic and germline activity. Furthermore, this assembly system is flexible and is compatible with standard molecular biology laboratory tools, but can be scaled with automated laboratory support. To demonstrate, we use a highly accessible and commercially available liquid-handling robot to rapidly and accurately assemble TALEs using the FusX TALE toolkit. Together, the FusX system accelerates TALE-based genomic science applications from basic science screening work for functional genomics testing and molecular medicine applications.

  20. Chitosan-Coated Magnetic Nanoparticles Prepared in One-Step by Precipitation in a High-Aqueous Phase Content Reverse Microemulsion

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    María Guadalupe Pineda

    2014-07-01

    Full Text Available Chitosan-coated magnetic nanoparticles (CMNP were prepared in one-step by precipitation in a high-aqueous phase content reverse microemulsion in the presence of chitosan. The high-aqueous phase concentration led to productivities close to 0.49 g CMNP/100 g microemulsion; much higher than those characteristic of precipitation in reverse microemulsions for preparing magnetic nanoparticles. The obtained nanoparticles present a narrow particle size distribution with an average diameter of 4.5 nm; appearing to be formed of a single crystallite; furthermore they present superparamagnetism and high magnetization values; close to 49 emu/g. Characterization of CMNP suggests that chitosan is present as a non-homogeneous very thin layer; which explains the slight reduction in the magnetization value of CMNP in comparison with that of uncoated magnetic nanoparticles. The prepared nanoparticles show high heavy ion removal capability; as demonstrated by their use in the treatment of Pb2+ aqueous solutions; from which lead ions were completely removed within 10 min.

  1. One-step microlithography

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    Kahlen, Franz-Josef; Sankaranarayanan, Srikanth; Kar, Aravinda

    1997-09-01

    Subject of this investigation is a one-step rapid machining process to create miniaturized 3D parts, using the original sample material. An experimental setup where metal powder is fed to the laser beam-material interaction region has been built. The powder is melted and forms planar, 2D geometries as the substrate is moved under the laser beam in XY- direction. After completing the geometry in the plane, the substrate is displaced in Z-direction, and a new layer of material is placed on top of the just completed deposit. By continuous repetition of this process, 3D parts wee created. In particular, the impact of the focal spot size of the high power laser beam on the smallest achievable structures was investigated. At a translation speed of 51 mm/s a minimum material thickness of 590 micrometers was achieved. Also, it was shown that a small Z-displacement has a negligible influence on the continuity of the material deposition over this power range. A high power CO2 laser was used as energy source, the material powder under investigation was stainless steel SS304L. Helium was used as shield gas at a flow rate of 15 1/min. The incident CO2 laser beam power was varied between 300 W and 400 W, with the laser beam intensity distribute in a donut mode. The laser beam was focused to a focal diameter of 600 (Mu) m.

  2. One-step separation and purification of two chromones and one pyrone from Aloe barbadensis Miller: a comparison between reversed-phase flash chromatography and high-speed counter current chromatography.

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    Zhong, Jia-Sheng; Wan, Jin-Zhi; Ding, Wen-Jing; Wu, Xiao-Fang; Xie, Zhi-Yong

    2014-01-01

    Chromones and pyrones are the major secondary metabolites of Aloe barbadensis Miller. As they are minor components of the plant, an efficient purification procedure for them is of great importance for promoting their pharmacological studies. To develop efficient methods for one-step separation and purification of two chromones (5-((S)-2'-oxo-4'-hydroxypentyl)-2-hydroxymethylchromone (1) and 5-((4E)-2'-oxo-pentenyl)-2-hydroxymethylchromone (3)) and one pyrone (aloenin aglycone (2)) from A. barbadensis via reversed-phase flash chromatography (RP-FC) and high-speed counter current chromatography (HSCCC). The RP-FC separation was performed using methanol:water (26:74, v/v) as the mobile phase at a flow rate of 20 mL/min. A solvent system composed of dichloromethane:methanol:water (3:1.5:1, v/v/v) was used for the HSCCC separation, at a flow rate of 2.0 mL/min. A one-step RP-FC operation within 110 min was successfully used for the purification of compounds 1 (27.9 mg, 96.5%), 2 (32.4 mg, 98.2%) and 3 (4.1 mg, 99.0%) from 129 mg of crude sample, and a one-step HSCCC separation within 95 min was successfully implemented for the purification of compounds 1 (31.1 mg, 97.6%), 2 (35.8 mg, 96.7%) and 3 (2.7 mg, 98.1%) from 134 mg of crude sample. The developed procedures were efficient, with low cost and high yield, which would afford sufficient amounts of high-purity compounds for chromatographic purposes and pharmacological activity screening. Copyright © 2014 John Wiley & Sons, Ltd.

  3. Reverse Transcription of Retroviruses and LTR Retrotransposons.

    Science.gov (United States)

    Hughes, Stephen H

    2015-04-01

    The enzyme reverse transcriptase (RT) was discovered in retroviruses almost 50 years ago. The demonstration that other types of viruses, and what are now called retrotransposons, also replicated using an enzyme that could copy RNA into DNA came a few years later. The intensity of the research in both the process of reverse transcription and the enzyme RT was greatly stimulated by the recognition, in the mid-1980s, that human immunodeficiency virus (HIV) was a retrovirus and by the fact that the first successful anti-HIV drug, azidothymidine (AZT), is a substrate for RT. Although AZT monotherapy is a thing of the past, the most commonly prescribed, and most successful, combination therapies still involve one or both of the two major classes of anti-RT drugs. Although the basic mechanics of reverse transcription were worked out many years ago, and the first high-resolution structures of HIV RT are now more than 20 years old, we still have much to learn, particularly about the roles played by the host and viral factors that make the process of reverse transcription much more efficient in the cell than in the test tube. Moreover, we are only now beginning to understand how various host factors that are part of the innate immunity system interact with the process of reverse transcription to protect the host-cell genome, the host cell, and the whole host, from retroviral infection, and from unwanted retrotransposition.

  4. The one step fermionic ladder

    Science.gov (United States)

    Das, Joy Prakash; Setlur, Girish S.

    2017-10-01

    The one step fermionic ladder refers to two parallel Luttinger Liquids (poles of the ladder) placed such that there is a finite probability of electrons hopping between the two poles at a pair of opposing points along each of the poles. The many-body Green function for such a system is calculated in presence of forward scattering interactions using the powerful non-chiral bosonization technique (NCBT). This technique is based on a non-standard harmonic analysis of the rapidly varying parts of the density fields appropriate for the study of strongly inhomogeneous ladder systems. The closed analytical expression for the correlation function obtained from NCBT is nothing but the series involving the RPA (Random Phase Approximation) diagrams in powers of the forward scattering coupling strength resummed to include only the most singular terms with the source of inhomogeneities treated exactly. Finally the correlation functions are used to study physical phenomena such as Friedel oscillations and the conductance of such systems with the potential difference applied across various ends.

  5. Cancer Cachexia: One Step Ahead.

    Science.gov (United States)

    Meriggi, Fausto

    2015-01-01

    Cachexia is one of the most common manifestations in advanced cancer patients, but too often it remains under- recognized and under-treated. Starvation is not the same of cachexia. Cachexia is defined by "weight loss >5% over past 6 months in absence of simple starvation or the combination of ongoing weight loss>2% with BMI cachexia is not fully understood, but inflammation and an increased catabolic response to a number of cancer-related factors seem to represent the basis of any assumption. Early diagnosis of a pre-cachectic or cachectic state is a key moment for the treatment of this complex syndrome, in order to guarantee an adequate food intake and suitable exercise and to interfere with the inflammatory processes that are typical of cachexia. Therefore, one of the main aims is to identify those patients most likely to develop the syndrome early. A multimodality baseline approach to cancer cachexia addresses reversible clinical contributory factors. There are currently no medicinal products that have a proven efficacy in the medical approach to cancer cachexia. Recently, anamorelin, a synthetic orally active ghrelin receptor agonist, showed promising results, but the best approach to cancer cachexia probably remains an early multimodal interventions consisting in nutritional intervention, exercise and rehabilitation program, and multi-target drug therapies. This review summarizes what we know and what still need to know about cancer cachexia syndrome.

  6. Screening of dengue virus in field-caught Aedes aegypti and Aedes albopictus (Diptera: Culicidae) by one-step SYBR green-based reverse transcriptase-polymerase chain reaction assay during 2004-2007 in Southern Taiwan.

    Science.gov (United States)

    Chen, Chien-Fu; Shu, Pei-Yun; Teng, Hwa-Jen; Su, Chien-Ling; Wu, Jhy-Wen; Wang, Jen-Hsin; Lin, Ting-Hsiang; Huang, Jyh-Hsiung; Wu, Ho-Sheng

    2010-12-01

    We carried out virological surveillance of dengue virus (DENV) in field-caught Aedes mosquitoes during 2004-2007 to estimate the monthly prevalence of infected females in dengue high-risk areas of Taiwan. A total of 92,892 Aedes aegypti (43,133 females and 49,759 males) and 79,315 Aedes albopictus (57,319 females and 21,996 males) adults were collected, grouped into 25,654 pools, and processed for virus detection using a one-step SYBR Green-based real-time reverse transcriptase-polymerase chain reaction assay. DENVs were periodically and sympatrically detected in Ae. aegypti females in accordance with major dengue outbreaks and the corresponding dengue serotypes. Only 0.2% of 7628 pools of Ae. aegypti females were positive for DENVs. This resulted in an overall estimated infection rate (maximum likelihood estimation) of 0.970 per 1000 mosquitoes (95% confidence interval [CI] = 0.53-1.65). The total monthly infection rates ranged from 0.50 to 2.23 per 1000 mosquitoes (95% CI = 0.03-10.71). When sampling areas were scaled down to the city level, monthly infection rates increased to 0.73-12.59 (95% CI = 0.06-59.19). Monthly infection rates over all sampling areas and at the city level increased significantly by month. All positive pools were collected in July (one pool), August (two pools), September (one pool), October (three pools), November (four pools), and December (one pool). All four virus serotypes were detected in mosquitoes, which were consistent with dengue serotypes infecting humans in 2004 (DENV-4), 2005 and 2006 (DENV-2 and DENV-3), and 2007 (DENV-1). Our results provide supporting evidence that, in general, DENV infection rates were low in local Aedes mosquito population during 2004-2007 and that transovarial transmission may not be occurring or is occurring at much lower rates than evidenced in some endemic countries.

  7. Reverse Transcriptase and Cellular Factors: Regulators of HIV-1 Reverse Transcription

    Directory of Open Access Journals (Sweden)

    David Harrich

    2009-11-01

    Full Text Available There is ample evidence that synthesis of HIV-1 proviral DNA from the viral RNA genome during reverse transcription requires host factors. However, only a few cellular proteins have been described in detail that affect reverse transcription and interact with reverse transcriptase (RT. HIV-1 integrase is an RT binding protein and a number of IN-binding proteins including INI1, components of the Sin3a complex, and Gemin2 affect reverse transcription. In addition, recent studies implicate the cellular proteins HuR, AKAP149, and DNA topoisomerase I in reverse transcription through an interaction with RT. In this review we will consider interactions of reverse transcription complex with viral and cellular factors and how they affect the reverse transcription process.

  8. From reverse transcription to human brain tumors

    Directory of Open Access Journals (Sweden)

    Dmitrenko V. V.

    2013-05-01

    Full Text Available Reverse transcriptase from avian myeloblastosis virus (AMV was the subject of the study, from which the investi- gations of the Department of biosynthesis of nucleic acids were started. Production of AMV in grams quantities and isolation of AMV reverse transcriptase were established in the laboratory during the seventies of the past cen- tury and this initiated research on the cDNA synthesis, cloning and investigation of the structure and functions of the eukaryotic genes. Structures of salmon insulin and insulin-like growth factor (IGF family genes and their transcripts were determined during long-term investigations. Results of two modern techniques, microarray-ba- sed hybridization and SAGE, were used for the identification of the genes differentially expressed in astrocytic gliomas and human normal brain. Comparison of SAGE results on the genes overexpressed in glioblastoma with the results of microarray analysis revealed a limited number of common genes. 105 differentially expressed genes, common to both methods, can be included in the list of candidates for the molecular typing of glioblastoma. The first experiments on the classification of glioblastomas based on the data of the 20 genes expression were conducted by using of artificial neural network analysis. The results of these experiments showed that the expression profiles of these genes in 224 glioblastoma samples and 74 normal brain samples could be according to the Koho- nen’s maps. The CHI3L1 and CHI3L2 genes of chitinase-like cartilage protein were revealed among the most overexpressed genes in glioblastoma, which could have prognostic and diagnostic potential. Results of in vitro experiments demonstrated that both proteins, CHI3L1 and CHI3L2, may initiate the phosphorylation of ERK1/ ERK2 and AKT kinases leading to the activation of MAPK/ERK1/2 and PI3K/AKT signaling cascades in human embryonic kidney 293 cells, human glioblastoma U87MG, and U373 cells. The new human cell line

  9. One-step fabrication of multifunctional micromotors

    Science.gov (United States)

    Gao, Wenlong; Liu, Mei; Liu, Limei; Zhang, Hui; Dong, Bin; Li, Christopher Y.

    2015-08-01

    Although artificial micromotors have undergone tremendous progress in recent years, their fabrication normally requires complex steps or expensive equipment. In this paper, we report a facile one-step method based on an emulsion solvent evaporation process to fabricate multifunctional micromotors. By simultaneously incorporating various components into an oil-in-water droplet, upon emulsification and solidification, a sphere-shaped, asymmetric, and multifunctional micromotor is formed. Some of the attractive functions of this model micromotor include autonomous movement in high ionic strength solution, remote control, enzymatic disassembly and sustained release. This one-step, versatile fabrication method can be easily scaled up and therefore may have great potential in mass production of multifunctional micromotors for a wide range of practical applications.Although artificial micromotors have undergone tremendous progress in recent years, their fabrication normally requires complex steps or expensive equipment. In this paper, we report a facile one-step method based on an emulsion solvent evaporation process to fabricate multifunctional micromotors. By simultaneously incorporating various components into an oil-in-water droplet, upon emulsification and solidification, a sphere-shaped, asymmetric, and multifunctional micromotor is formed. Some of the attractive functions of this model micromotor include autonomous movement in high ionic strength solution, remote control, enzymatic disassembly and sustained release. This one-step, versatile fabrication method can be easily scaled up and therefore may have great potential in mass production of multifunctional micromotors for a wide range of practical applications. Electronic supplementary information (ESI) available: Videos S1-S4 and Fig. S1-S3. See DOI: 10.1039/c5nr03574k

  10. Novel Rotavirus VP7 Typing Assay Using a One-Step Reverse Transcriptase PCR Protocol and Product Sequencing and Utility of the Assay for Epidemiological Studies and Strain Characterization, Including Serotype Subgroup Analysis

    Science.gov (United States)

    DiStefano, Daniel J.; Kraiouchkine, Nikolai; Mallette, Laura; Maliga, Marianne; Kulnis, Gregory; Keller, Paul M.; Clark, H. Fred; Shaw, Alan R.

    2005-01-01

    Rotavirus is the most common cause of severe dehydrating gastroenteritis in infants. To date, 10 different serotypes of rotavirus have been identified in human stools. While four or five serotypes dominate, serotype circulation varies with season and geography. Since our laboratory has been involved in the development of a multivalent rotavirus vaccine, it is important to identify the serotypes of rotavirus encountered during our clinical trials. We have developed methodologies for the molecular identification of rotavirus strains based on VP7 gene segment sequence. A 365-bp reverse transcriptase PCR product was generated from the VP7 gene segment using a pair of novel degenerate primers. All serotypes tested (both animal and human) yielded an identically sized product after amplification. Sequencing of these products is performed using truncated versions of the original primers. The sequence generated is compared against a database of rotavirus VP7 sequences, with the G type determined, based on the sequence homology. Using this assay, we have correctly identified human VP7 strains from a panel of available serotypes, as well as numerous animal strains. The assay was qualified using rotavirus positive stool samples, negative stool samples, and rotavirus-spiked stool samples. In addition, samples from cases of acute gastroenteritis collected at Children's Hospital of Philadelphia have been evaluated and indicate that the assay is able to discriminate subtle differences within serotypes. The assay has been utilized in the testing of >3,000 antigen-positive (enzyme immunoassay) samples collected during clinical trials of a rotavirus vaccine (RotaTeq) and identified a serotype in ∼92% of samples (3, 17, 19). PMID:16333070

  11. One-step lowrank wave extrapolation

    KAUST Repository

    Sindi, Ghada Atif

    2014-01-01

    Wavefield extrapolation is at the heart of modeling, imaging, and Full waveform inversion. Spectral methods gained well deserved attention due to their dispersion free solutions and their natural handling of anisotropic media. We propose a scheme a modified one-step lowrank wave extrapolation using Shanks transform in isotropic, and anisotropic media. Specifically, we utilize a velocity gradient term to add to the accuracy of the phase approximation function in the spectral implementation. With the higher accuracy, we can utilize larger time steps and make the extrapolation more efficient. Applications to models with strong inhomogeneity and considerable anisotropy demonstrates the utility of the approach.

  12. Transcription activator-like effector nuclease (TALEN-mediated CLYBL targeting enables enhanced transgene expression and one-step generation of dual reporter human induced pluripotent stem cell (iPSC and neural stem cell (NSC lines.

    Directory of Open Access Journals (Sweden)

    Trevor Cerbini

    Full Text Available Targeted genome engineering to robustly express transgenes is an essential methodology for stem cell-based research and therapy. Although designer nucleases have been used to drastically enhance gene editing efficiency, targeted addition and stable expression of transgenes to date is limited at single gene/locus and mostly PPP1R12C/AAVS1 in human stem cells. Here we constructed transcription activator-like effector nucleases (TALENs targeting the safe-harbor like gene CLYBL to mediate reporter gene integration at 38%-58% efficiency, and used both AAVS1-TALENs and CLYBL-TALENs to simultaneously knock-in multiple reporter genes at dual safe-harbor loci in human induced pluripotent stem cells (iPSCs and neural stem cells (NSCs. The CLYBL-TALEN engineered cell lines maintained robust reporter expression during self-renewal and differentiation, and revealed that CLYBL targeting resulted in stronger transgene expression and less perturbation on local gene expression than PPP1R12C/AAVS1. TALEN-mediated CLYBL engineering provides improved transgene expression and options for multiple genetic modification in human stem cells.

  13. Transcription activator-like effector nuclease (TALEN)-mediated CLYBL targeting enables enhanced transgene expression and one-step generation of dual reporter human induced pluripotent stem cell (iPSC) and neural stem cell (NSC) lines.

    Science.gov (United States)

    Cerbini, Trevor; Funahashi, Ray; Luo, Yongquan; Liu, Chengyu; Park, Kyeyoon; Rao, Mahendra; Malik, Nasir; Zou, Jizhong

    2015-01-01

    Targeted genome engineering to robustly express transgenes is an essential methodology for stem cell-based research and therapy. Although designer nucleases have been used to drastically enhance gene editing efficiency, targeted addition and stable expression of transgenes to date is limited at single gene/locus and mostly PPP1R12C/AAVS1 in human stem cells. Here we constructed transcription activator-like effector nucleases (TALENs) targeting the safe-harbor like gene CLYBL to mediate reporter gene integration at 38%-58% efficiency, and used both AAVS1-TALENs and CLYBL-TALENs to simultaneously knock-in multiple reporter genes at dual safe-harbor loci in human induced pluripotent stem cells (iPSCs) and neural stem cells (NSCs). The CLYBL-TALEN engineered cell lines maintained robust reporter expression during self-renewal and differentiation, and revealed that CLYBL targeting resulted in stronger transgene expression and less perturbation on local gene expression than PPP1R12C/AAVS1. TALEN-mediated CLYBL engineering provides improved transgene expression and options for multiple genetic modification in human stem cells.

  14. Reverse transcription of the HIV-1 pandemic.

    Science.gov (United States)

    Basavapathruni, Aravind; Anderson, Karen S

    2007-12-01

    The HIV/AIDS pandemic has existed for >25 years. Extensive work globally has provided avenues to combat viral infection, but the disease continues to rage on in the human population and infected approximately 4 million people in 2006 alone. In this review, we provide a brief history of HIV/AIDS, followed by analysis of one therapeutic target of HIV-1: its reverse transcriptase (RT). We discuss the biochemical characterization of RT in order to place emphasis on possible avenues of inhibition, which now includes both nucleoside and non-nucleoside modalities. Therapies against RT remain a cornerstone of anti-HIV treatment, but the virus eventually resists inhibition through the selection of drug-resistant RT mutations. Current inhibitors and associated resistance are discussed, with the hopes that new therapeutics can be developed against RT.

  15. Transcriptional Regulation of Telomerase Reverse Transcriptase (TERT) by MYC

    Science.gov (United States)

    Khattar, Ekta; Tergaonkar, Vinay

    2017-01-01

    Telomerase elongates telomeres and is crucial for maintaining genomic stability. While stem cells and cancer cells display high telomerase activity, normal somatic cells lack telomerase activity primarily due to transcriptional repression of telomerase reverse transcriptase (TERT), the catalytic component of telomerase. Transcription factor binding, chromatin status as well as epigenetic modifications at the TERT promoter regulates TERT transcription. Myc is an important transcriptional regulator of TERT that directly controls its expression by promoter binding and associating with other transcription factors. In this review, we discuss the current understanding of the molecular mechanisms behind regulation of TERT transcription by Myc. We also discuss future perspectives in investigating the regulation of Myc at TERT promoter during cancer development.

  16. Detection of Papaya leaf distortion mosaic virus by reverse-transcription loop-mediated isothermal amplification.

    Science.gov (United States)

    Shen, Wentao; Tuo, Decai; Yan, Pu; Li, Xiaoying; Zhou, Peng

    2014-01-01

    Papaya leaf distortion mosaic virus (PLDMV) can infect transgenic papaya resistant to a related pathogen, Papaya ringspot virus (PRSV), posing a substantial threat to papaya production in China. Current detection methods, however, are unable to be used for rapid detection in the field. Here, a reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of PLDMV, using a set of four RT-LAMP primers designed based on the conserved sequence of PLDMV CP. The RT-LAMP method detected specifically PLDMV and was highly sensitive, with a detection limit of 1.32×10(-6) μg of total RNA per reaction. Indeed, the reaction was 10 times more sensitive than one-step RT-PCR, while also requiring significantly less time and equipment. The effectiveness of RT-LAMP and one-step RT-PCR in detecting the virus were compared using 90 field samples of non-transgenic papaya and 90 field samples of commercialized PRSV-resistant transgenic papaya from Hainan Island. None of the non-transgenic papaya tested positive for PLDMV using either method. In contrast, 19 of the commercialized PRSV-resistant transgenic papaya samples tested positive by RT-LAMP assay, and 6 of those tested negative by RT-PCR. Therefore, the PLDMV-specific RT-LAMP is a simple, rapid, sensitive, and cost-effective tool in the field diagnosis and control of PLDMV.

  17. 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...

  18. Combined in vitro transcription and reverse transcription to amplify and label complex synthetic oligonucleotide probe libraries.

    Science.gov (United States)

    Murgha, Yusuf; Beliveau, Brian; Semrau, Kassandra; Schwartz, Donald; Wu, Chao-Ting; Gulari, Erdogan; Rouillard, Jean-Marie

    2015-06-01

    Oligonucleotide microarrays allow the production of complex custom oligonucleotide libraries for nucleic acid detection-based applications such as fluorescence in situ hybridization (FISH). We have developed a PCR-free method to make single-stranded DNA (ssDNA) fluorescent probes through an intermediate RNA library. A double-stranded oligonucleotide library is amplified by transcription to create an RNA library. Next, dye- or hapten-conjugate primers are used to reverse transcribe the RNA to produce a dye-labeled cDNA library. Finally the RNA is hydrolyzed under alkaline conditions to obtain the single-stranded fluorescent probes library. Starting from unique oligonucleotide library constructs, we present two methods to produce single-stranded probe libraries. The two methods differ in the type of reverse transcription (RT) primer, the incorporation of fluorescent dye, and the purification of fluorescent probes. The first method employs dye-labeled reverse transcription primers to produce multiple differentially single-labeled probe subsets from one microarray library. The fluorescent probes are purified from excess primers by oligonucleotide-bead capture. The second method uses an RNA:DNA chimeric primer and amino-modified nucleotides to produce amino-allyl probes. The excess primers and RNA are hydrolyzed under alkaline conditions, followed by probe purification and labeling with amino-reactive dyes. The fluorescent probes created by the combination of transcription and reverse transcription can be used for FISH and to detect any RNA and DNA targets via hybridization.

  19. Resetting the transcription factor network reverses terminal chronic hepatic failure.

    Science.gov (United States)

    Nishikawa, Taichiro; Bell, Aaron; Brooks, Jenna M; Setoyama, Kentaro; Melis, Marta; Han, Bing; Fukumitsu, Ken; Handa, Kan; Tian, Jianmin; Kaestner, Klaus H; Vodovotz, Yoram; Locker, Joseph; Soto-Gutierrez, Alejandro; Fox, Ira J

    2015-04-01

    The cause of organ failure is enigmatic for many degenerative diseases, including end-stage liver disease. Here, using a CCl4-induced rat model of irreversible and fatal hepatic failure, which also exhibits terminal changes in the extracellular matrix, we demonstrated that chronic injury stably reprograms the critical balance of transcription factors and that diseased and dedifferentiated cells can be returned to normal function by re-expression of critical transcription factors, a process similar to the type of reprogramming that induces somatic cells to become pluripotent or to change their cell lineage. Forced re-expression of the transcription factor HNF4α induced expression of the other hepatocyte-expressed transcription factors; restored functionality in terminally diseased hepatocytes isolated from CCl4-treated rats; and rapidly reversed fatal liver failure in CCl4-treated animals by restoring diseased hepatocytes rather than replacing them with new hepatocytes or stem cells. Together, the results of our study indicate that disruption of the transcription factor network and cellular dedifferentiation likely mediate terminal liver failure and suggest reinstatement of this network has therapeutic potential for correcting organ failure without cell replacement.

  20. APOBEC3G inhibits elongation of HIV-1 reverse transcripts.

    Directory of Open Access Journals (Sweden)

    Kate N Bishop

    2008-12-01

    Full Text Available APOBEC3G (A3G is a host cytidine deaminase that, in the absence of Vif, restricts HIV-1 replication and reduces the amount of viral DNA that accumulates in cells. Initial studies determined that A3G induces extensive mutation of nascent HIV-1 cDNA during reverse transcription. It has been proposed that this triggers the degradation of the viral DNA, but there is now mounting evidence that this mechanism may not be correct. Here, we use a natural endogenous reverse transcriptase assay to show that, in cell-free virus particles, A3G is able to inhibit HIV-1 cDNA accumulation not only in the absence of hypermutation but also without the apparent need for any target cell factors. We find that although reverse transcription initiates in the presence of A3G, elongation of the cDNA product is impeded. These data support the model that A3G reduces HIV-1 cDNA levels by inhibiting synthesis rather than by inducing degradation.

  1. One-Step RT-PCR protocols improve the rate of dengue diagnosis compared to Two-Step RT-PCR approaches.

    Science.gov (United States)

    De Paula, Sérgio Oliveira; de Melo Lima, Cristiane; Torres, Maria Paula; Pereira, Márcia Rodrigues Garbin; Lopes da Fonseca, Benedito Antônio

    2004-08-01

    Dengue is the most important arboviral disease transmitted to humans. In our laboratory, we have been working on the standardization of the polymerase chain reaction (PCR) diagnosis of this disease. In this work, we compared five commercial kits regularly used on reverse-transcription polymerase chain reaction (RT-PCR) protocols: two Two-Step kits (SuperScript II RT/Super Mix kit and reverse transcription system/Taq DNA polymerase) and three One-Step kits (ready-to-go RT-PCR Beads kit, QIAGEN One-Step RT-PCR kit, and AcessQuick RT-PCR system). Thirty-one serum samples of patients with clinical diagnosis of dengue fever (DF) were analyzed by RT-PCR and serology. RNA extraction was done with the QIAamp Viral RNA kit, and cDNA synthesis and PCR done according to the manufacturer's protocol for the five kits. Out of the 31 serum samples collected from patients suspected of having dengue, 27 were IgM-positive, confirming the dengue diagnosis. Out of those, 24 were positive by the ready-to-go RT-PCR Beads kit, 25 were positive by AcessQuick RT-PCR system and 27 were positive by QIAGEN One-Step RT-PCR kit. On the other hand, only six samples were positive by the SuperScript II RT/Super Mix kits and 10 were positive by reverse transcription system/Taq DNA polymerase kit. The best performance observed with the One-Step kits was confirmed in spiked samples with known quantities of dengue-1 virus since they detected up 1 x 10(2) PFU/ml, while the most sensitive Two-Step kit detected up 1 x 10(4) PFU/ml. These data show that One-Step RT-PCR kits yielded a higher rate of dengue virus detection than the Two-Step kits and correlated well with the serological diagnosis.

  2. Reverse protection assay: a tool to analyze transcriptional rates from individual promoters.

    Science.gov (United States)

    Zubo, Yan O; Kusnetsov, Victor V; Börner, Thomas; Liere, Karsten

    2011-12-20

    Transcriptional activity of entire genes in chloroplasts is usually assayed by run-on analyses. To determine not only the overall intensity of transcription of a gene, but also the rate of transcription from a particular promoter, we created the Reverse RNase Protection Assay (RePro): in-organello run-on transcription coupled to RNase protection to define distinct transcript ends during transcription. We demonstrate successful application of RePro in plastid promoter analysis and transcript 3' end processing.

  3. Reverse transcription loop-mediated isothermal amplification assay for rapid detection of Papaya ringspot virus.

    Science.gov (United States)

    Shen, Wentao; Tuo, Decai; Yan, Pu; Yang, Yong; Li, Xiaoying; Zhou, Peng

    2014-08-01

    Papaya ringspot virus (PRSV) and Papaya leaf distortion mosaic virus (PLDMV), which causes disease symptoms similar to PRSV, threaten commercial production of both non-transgenic-papaya and PRSV-resistant transgenic papaya in China. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay to detect PLDMV was developed previously. In this study, the development of another RT-LAMP assay to distinguish among transgenic, PRSV-infected and PLDMV-infected papaya by detection of PRSV is reported. A set of four RT-LAMP primers was designed based on the highly conserved region of the P3 gene of PRSV. The RT-LAMP method was specific and sensitive in detecting PRSV, with a detection limit of 1.15×10(-6)μg of total RNA per reaction. Indeed, the reaction was 10 times more sensitive than one-step RT-PCR. Field application of the RT-LAMP assay demonstrated that samples positive for PRSV were detected only in non-transgenic papaya, whereas samples positive for PLDMV were detected only in commercialized PRSV-resistant transgenic papaya. This suggests that PRSV remains the major limiting factor for non-transgenic-papaya production, and the emergence of PLDMV threatens the commercial transgenic cultivar in China. However, this study, combined with the earlier development of an RT-LAMP assay for PLDMV, will provide a rapid, sensitive and cost-effective diagnostic power to distinguish virus infections in papaya.

  4. Critical analysis of rhinovirus RNA load quantification by real-time reverse transcription-PCR.

    Science.gov (United States)

    Schibler, Manuel; Yerly, Sabine; Vieille, Gaël; Docquier, Mylène; Turin, Lara; Kaiser, Laurent; Tapparel, Caroline

    2012-09-01

    Rhinoviruses are the most frequent cause of human respiratory infections, and quantitative rhinovirus diagnostic tools are needed for clinical investigations. Although results obtained by real-time reverse-transcription PCR (RT-PCR) assays are frequently converted to viral RNA loads, this presents several limitations regarding accurate virus RNA quantification, particularly given the need to reliably quantify all known rhinovirus genotypes with a single assay. Using an internal extraction control and serial dilutions of an in vitro-transcribed rhinovirus RNA reference standard, we validated a quantitative one-step real-time PCR assay. We then used chimeric rhinovirus genomes with 5'-untranslated regions (5'UTRs) originating from the three rhinovirus species and from one enterovirus to estimate the impact of the 5'UTR diversity. Respiratory specimens from infected patients were then also analyzed. The assay quantification ability ranged from 4.10 to 9.10 log RNA copies/ml, with an estimated error margin of ±10%. This variation was mainly linked to target variability and interassay variability. Taken together, our results indicate that our assay can reliably estimate rhinovirus RNA load, provided that the appropriate error margin is used. In contrast, due to the lack of a universal rhinovirus RNA standard and the variability related to sample collection procedures, accurate absolute rhinovirus RNA quantification in respiratory specimens is currently hardly feasible.

  5. Transcription, reverse transcription, and analysis of RNA containing artificial genetic components.

    Science.gov (United States)

    Leal, Nicole A; Kim, Hyo-Joong; Hoshika, Shuichi; Kim, Myong-Jung; Carrigan, Matthew A; Benner, Steven A

    2015-04-17

    Expanding the synthetic biology of artificially expanded genetic information systems (AEGIS) requires tools to make and analyze RNA molecules having added nucleotide "letters". We report here the development of T7 RNA polymerase and reverse transcriptase to catalyze transcription and reverse transcription of xNA (DNA or RNA) having two complementary AEGIS nucleobases, 6-amino-5-nitropyridin-2-one (trivially, Z) and 2-aminoimidazo[1,2a]-1,3,5-triazin-4(8H)-one (trivially, P). We also report MALDI mass spectrometry and HPLC-based analyses for oligomeric GACUZP six-letter RNA and the use of ribonuclease (RNase) A and T1 RNase as enzymatic tools for the sequence-specific degradation of GACUZP RNA. We then applied these tools to analyze the GACUZP and GACTZP products of polymerases and reverse transcriptases (respectively) made from DNA and RNA templates. In addition to advancing this 6-letter AEGIS toward the biosynthesis of proteins containing additional amino acids, these experiments provided new insights into the biophysics of DNA.

  6. Rapid detection of peste des petits ruminants virus by a reverse transcription loop-mediated isothermal amplification assay.

    Science.gov (United States)

    Li, Lin; Bao, Jingyue; Wu, Xiaodong; Wang, Zhiliang; Wang, Junwei; Gong, Mingxia; Liu, Chunju; Li, Jinming

    2010-12-01

    Peste des petits ruminants virus (PPRV) is the causative agent of peste des petits ruminants (PPR), an economically important viral disease of small ruminants. In this report, a one-step, single-tube, reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of PPRV. A set of six LAMP primers were designed based on the matrix gene sequence of PPRV to amplify the target RNA by incubation at 63°C for 60min with Bst DNA polymerase and reverse transcriptase. The amplified products could be observed by the naked eye. The specificity of the RT-LAMP assay was validated by amplifying eight strains of PPRV isolated in different geographical areas. No cross-reactivity with other related viruses, including rinderpest virus, canine distemper virus and measles virus, was detected. The sensitivity of the assay was similar to that of real-time reverse transcription polymerase chain reaction (RT-PCR) and 10-fold higher than that of conventional RT-PCR. Twenty clinical samples were evaluated by the RT-LAMP assay, and the results were consistent with those of real-time RT-PCR. As a simple, rapid and accurate detection method, this RT-LAMP assay has important potential applications in the clinical diagnosis of PPR and the surveillance of PPRV. Copyright © 2010 Elsevier B.V. All rights reserved.

  7. Properties of the reverse transcription reaction in mRNA quantification

    DEFF Research Database (Denmark)

    Ståhlberg, Anders; Håkansson, Joakim; Xian, Xiaojie;

    2004-01-01

    BACKGROUND: In most measurements of gene expression, mRNA is first reverse-transcribed into cDNA. We studied the reverse transcription reaction and its consequences for quantitative measurements of gene expression. METHODS: We used SYBR green I-based quantitative real-time PCR (QPCR) to measure......-QPCR) was mainly attributable to the reverse transcription step. Reverse transcription efficiency depended on priming strategy, and the dependence was different for the five genes studied. Reverse transcription yields also depended on total RNA concentration. CONCLUSIONS: RT-QPCR gene expression measurements...... are comparable only when the same priming strategy and reaction conditions are used in all experiments and the samples contain the same total amount of RNA. Experimental accuracy is improved by running samples in (at least) duplicate starting with the reverse transcription reaction....

  8. Design and optimization of reverse-transcription quantitative PCR experiments.

    Science.gov (United States)

    Tichopad, Ales; Kitchen, Rob; Riedmaier, Irmgard; Becker, Christiane; Ståhlberg, Anders; Kubista, Mikael

    2009-10-01

    Quantitative PCR (qPCR) is a valuable technique for accurately and reliably profiling and quantifying gene expression. Typically, samples obtained from the organism of study have to be processed via several preparative steps before qPCR. We estimated the errors of sample withdrawal and extraction, reverse transcription (RT), and qPCR that are introduced into measurements of mRNA concentrations. We performed hierarchically arranged experiments with 3 animals, 3 samples, 3 RT reactions, and 3 qPCRs and quantified the expression of several genes in solid tissue, blood, cell culture, and single cells. A nested ANOVA design was used to model the experiments, and relative and absolute errors were calculated with this model for each processing level in the hierarchical design. We found that intersubject differences became easily confounded by sample heterogeneity for single cells and solid tissue. In cell cultures and blood, the noise from the RT and qPCR steps contributed substantially to the overall error because the sampling noise was less pronounced. We recommend the use of sample replicates preferentially to any other replicates when working with solid tissue, cell cultures, and single cells, and we recommend the use of RT replicates when working with blood. We show how an optimal sampling plan can be calculated for a limited budget. .

  9. Evaluation of one-step luminescent cyanoacrylate fuming.

    Science.gov (United States)

    Khuu, Alicia; Chadwick, Scott; Spindler, Xanthe; Lam, Rolanda; Moret, Sébastien; Roux, Claude

    2016-06-01

    One-step luminescent cyanoacrylates have recently been introduced as an alternative to the conventional cyanoacrylate fuming methods. These new techniques do not require the application of a luminescent post-treatment in order to enhance cyanoacrylate-developed fingermarks. In this study, three one-step polymer cyanoacrylates: CN Yellow Crystals (Aneval Inc.), PolyCyano UV (Foster+Freeman Ltd.) and PECA Multiband (BVDA), and one monomer cyanoacrylate: Lumikit™ (Crime Scene Technology), were evaluated against a conventional two-step cyanoacrylate fuming method (Cyanobloom (Foster+Freeman Ltd.) with rhodamine 6G stain). The manufacturers' recommended conditions or conditions compatible with the MVC™ 1000/D (Foster+Freeman Ltd.) were assessed with fingermarks aged for up to 8 weeks on non-porous and semi-porous substrates. Under white light, Cyanobloom generally gave better development than the one-step treatments across the substrates. Similarly when viewed under the respective luminescent conditions, Cyanobloom with rhodamine 6G stain resulted in improved contrast against the one-step treatments except on polystyrene, where PolyCyano UV and PECA Multiband gave better visualisation. Rhodamine 6G post-treatment of one-step samples did not significantly enhance the contrast of any of the one-step treatments against Cyanobloom/rhodamine 6G-treated samples.

  10. HuR interacts with human immunodeficiency virus type 1 reverse transcriptase, and modulates reverse transcription in infected cells

    Directory of Open Access Journals (Sweden)

    Ennifar Eric

    2008-06-01

    Full Text Available Abstract Reverse transcription of the genetic material of human immunodeficiency virus type 1 (HIV-1 is a critical step in the replication cycle of this virus. This process, catalyzed by reverse transcriptase (RT, is well characterized at the biochemical level. However, in infected cells, reverse transcription occurs in a multiprotein complex – the reverse transcription complex (RTC – consisting of viral genomic RNA associated with viral proteins (including RT and, presumably, as yet uncharacterized cellular proteins. Very little is known about the cellular proteins interacting with the RTC, and with reverse transcriptase in particular. We report here that HIV-1 reverse transcription is affected by the levels of a nucleocytoplasmic shuttling protein – the RNA-binding protein HuR. A direct protein-protein interaction between RT and HuR was observed in a yeast two-hybrid screen and confirmed in vitro by homogenous time-resolved fluorescence (HTRF. We mapped the domain interacting with HuR to the RNAse H domain of RT, and the binding domain for RT to the C-terminus of HuR, partially overlapping the third RRM RNA-binding domain of HuR. HuR silencing with specific siRNAs greatly impaired early and late steps of reverse transcription, significantly inhibiting HIV-1 infection. Moreover, by mutagenesis and immunoprecipitation studies, we could not detect the binding of HuR to the viral RNA. These results suggest that HuR may be involved in and may modulate the reverse transcription reaction of HIV-1, by an as yet unknown mechanism involving a protein-protein interaction with HIV-1 RT.

  11. Reverse protection assay: a tool to analyze transcriptional rates from individual promoters

    Directory of Open Access Journals (Sweden)

    Zubo Yan O

    2011-12-01

    Full Text Available Abstract Transcriptional activity of entire genes in chloroplasts is usually assayed by run-on analyses. To determine not only the overall intensity of transcription of a gene, but also the rate of transcription from a particular promoter, we created the Reverse RNase Protection Assay (RePro: in-organello run-on transcription coupled to RNase protection to define distinct transcript ends during transcription. We demonstrate successful application of RePro in plastid promoter analysis and transcript 3' end processing.

  12. Variable bandwidth and one-step local M-estimator

    Institute of Scientific and Technical Information of China (English)

    范剑青; 蒋建成

    2000-01-01

    A robust version of local linear regression smoothers augmented with variable bandwidth is studied. The proposed method inherits the advantages of local polynomial regression and overcomes the shortcoming of lack of robustness of least-squares techniques. The use of variable bandwidth enhances the flexibility of the resulting local M- estimators and makes them possible to cope well with spatially inho-mogeneous curves, heteroscedastic errors and nonuniform design densities. Under appropriate regularity conditions, it is shown that the proposed estimators exist and are asymptotically normal. Based on the robust estimation equation, one-step local M-estimators are introduced to reduce computational burden. It is demonstrated that the one-step local M-estimators share the same asymptotic distributions as the fully iterative M-estimators, as long as the initial estimators are good enough. In other words, the one-step local M-estimators reduce significantly the computation cost of the fully iterative M-estim

  13. One-step fabrication of supramolecular microcapsules from microfluidic droplets.

    Science.gov (United States)

    Zhang, Jing; Coulston, Roger J; Jones, Samuel T; Geng, Jin; Scherman, Oren A; Abell, Chris

    2012-02-10

    Although many techniques exist for preparing microcapsules, it is still challenging to fabricate them in an efficient and scalable process without compromising functionality and encapsulation efficiency. We demonstrated a simple one-step approach that exploits a versatile host-guest system and uses microfluidic droplets to generate porous microcapsules with easily customizable functionality. The capsules comprise a polymer-gold nanoparticle composite held together by cucurbit[8]uril ternary complexes. The dynamic yet highly stable micrometer-sized structures can be loaded in one step during capsule formation and are amenable to on-demand encapsulant release. The internal chemical environment can be probed with surface enhanced Raman spectroscopy.

  14. Convenient one-step synthesis of 5-carboxy-seminaphthofluoresceins

    DEFF Research Database (Denmark)

    Hammershøj, Peter; Thyhaug, Erling; Harris, Pernille

    2017-01-01

    The one-step synthesis and characterization of a series of regioisomerically pure 5-carboxy-seminaphthofluoresceins (5-carboxy-SNAFLs) is reported. The optical properties were determined in aqueous buffer at around biological pH, and highly pH sensitive, large Stokes-shift fluorophores with emiss......The one-step synthesis and characterization of a series of regioisomerically pure 5-carboxy-seminaphthofluoresceins (5-carboxy-SNAFLs) is reported. The optical properties were determined in aqueous buffer at around biological pH, and highly pH sensitive, large Stokes-shift fluorophores...

  15. Fundamental study of a one-step ambient temperature ferrite ...

    African Journals Online (AJOL)

    Fundamental study of a one-step ambient temperature ferrite process for treatment ... The approach involves the controlled oxidation of ferrous-containing AMD water at ... The resulting oxidation product is the ferrite (M13+2M22+O4) magnetite ...

  16. Convenient one-step synthesis of 5-carboxy-seminaphthofluoresceins

    DEFF Research Database (Denmark)

    Hammershøj, Peter; Thyhaug, Erling; Harris, Pernille

    2017-01-01

    The one-step synthesis and characterization of a series of regioisomerically pure 5-carboxy-seminaphthofluoresceins (5-carboxy-SNAFLs) is reported. The optical properties were determined in aqueous buffer at around biological pH, and highly pH sensitive, large Stokes-shift fluorophores with emiss...

  17. Single-Reaction Multiplex Reverse Transcription PCR for Detection of Zika, Chikungunya, and Dengue Viruses.

    Science.gov (United States)

    Waggoner, Jesse J; Gresh, Lionel; Mohamed-Hadley, Alisha; Ballesteros, Gabriela; Davila, Maria Jose Vargas; Tellez, Yolanda; Sahoo, Malaya K; Balmaseda, Angel; Harris, Eva; Pinsky, Benjamin A

    2016-07-01

    Clinical manifestations of Zika virus, chikungunya virus, and dengue virus infections can be similar. To improve virus detection, streamline molecular workflow, and decrease test costs, we developed and evaluated a multiplex real-time reverse transcription PCR for these viruses.

  18. Single-Reaction Multiplex Reverse Transcription PCR for Detection of Zika, Chikungunya, and Dengue Viruses

    OpenAIRE

    Waggoner, Jesse J.; Gresh, Lionel; Mohamed-Hadley, Alisha; Ballesteros, Gabriela; Davila, Maria Jose Vargas; Tellez, Yolanda; Sahoo, Malaya K.; Balmaseda, Angel; Harris, Eva; Pinsky, Benjamin A.

    2016-01-01

    Clinical manifestations of Zika virus, chikungunya virus, and dengue virus infections can be similar. To improve virus detection, streamline molecular workflow, and decrease test costs, we developed and evaluated a multiplex real-time reverse transcription PCR for these viruses.

  19. One-step pickling-activation before magnesium alloy plating

    Institute of Scientific and Technical Information of China (English)

    WANG Xin-juan; YU Gang; OUYANG Yue-jun; HE Xiao-mei; ZHANG Jun; YE Li-yuan

    2009-01-01

    A one-step pickling-activation process was proposed as an environmental friendly pretreatment method in phosphate-permanganate solution before electroplating on magnesium alloys. The effects of pickling-activation on qualities of coating were assessed by adhesion and porosity testing of copper plating. The interfacial reactions between specimen and solution were analyzed with SEM, EDX and XRD. The results show that the developed process of pickling-activation can equalize the potentials on substrate surface. The compacted zinc film can be obtained by zinc immersion after treating magnesium alloy in the pH 4-6 phosphate-permanganate solution for 3-5 min. The adhesion and corrosion resistance of copper plating are enhanced. The one-step pickling-activation can replace the existing two-step process of acid pickling and activation which contains a great deal of chromium and fluorine. The procedure of surface pretreatment is simplified and the production environment is improved.

  20. A novel one-step Helicobacter pylori saliva antigen test

    Directory of Open Access Journals (Sweden)

    Bi-Ling Yang

    2015-02-01

    Conclusion: The one-step HPS test exhibited a high sensitivity and low specificity compared with the other tests, indicating that it is not sufficiently accurate for use in a clinical setting for diagnosing H. pylori infection. However, the test is simple to use (requiring only a saliva sample, inexpensive, and noninvasive in its application, and thus appealing for use in population-based prevalence surveys of the epidemiology of H. pylori infection.

  1. Variable bandwidth and one-step local M-estimator

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    A robust version of local linear regression smoothers augmented with variable bandwidth is studied. The proposed method inherits the advantages of local polynomial regression and overcomes the shortcoming of lack of robustness of least-squares techniques. The use of variable bandwidth enhances the flexibility of the resulting local M-estimators and makes them possible to cope well with spatially inhomogeneous curves, heteroscedastic errors and nonuniform design densities. Under appropriate regularity conditions, it is shown that the proposed estimators exist and are asymptotically normal. Based on the robust estimation equation, one-step local M-estimators are introduced to reduce computational burden. It is demonstrated that the one-step local M-estimators share the same asymptotic distributions as the fully iterative M-estimators, as long as the initial estimators are good enough. In other words, the one-step local M-estimators reduce significantly the computation cost of the fully iterative M-estimators without deteriorating their performance. This fact is also illustrated via simulations.

  2. A Class of Multiderivative Hybrid One-step Methods

    Institute of Scientific and Technical Information of China (English)

    Feng-jian Yang; Xin-ming Chen; Ai-guo Zhang

    2002-01-01

    This paper presents a class of hybrid one-step methods that are obtained by using Cramer's rule and rational approximations to function exp (q). The algorithms fall into the catalogue of implicit formula, which involves sth order derivative and s+1 free parameters. The order of the algorithms satisfies s+1≤p≤2s+2. The stability of the methods is also studied, necessary and sufficient conditions for A-stability and L-stability are given. In addition, some examples are also given to demonstrate the method presented.

  3. One-step 18F labeling of biomolecules using organotrifluoroborates

    Science.gov (United States)

    Liu, Zhibo; Lin, Kuo-Shyan; Bénard, François; Pourghiasian, Maral; Kiesewetter, Dale O; Perrin, David M; Chen, Xiaoyuan

    2017-01-01

    Herein we present a general protocol for the functionalization of biomolecules with an organotrifluoroborate moiety so that they can be radiolabeled with aqueous 18F fluoride (18F−) and used for positron emission tomography (PET) imaging. Among the β+-emitting radionuclides, fluorine-18 (18F) is the isotope of choice for PET, and it is produced, on-demand, in many hospitals worldwide. Organotrifluoroborates can be 18F-labeled in one step in aqueous conditions via 18F–19F isotope exchange. This protocol features a recently designed ammoniomethyltrifluoroborate, and it describes the following: (i) a synthetic strategy that affords modular synthesis of radiolabeling precursors via a copper-catalyzed ‘click’ reaction; and (ii) a one-step 18F-labeling method that obviates the need for HPLC purification. Within 30 min, 18F-labeled PET imaging probes, such as peptides, can be synthesized in good chemical and radiochemical purity (>98%), satisfactory radiochemical yield of 20–35% (n > 20, non-decay corrected) and high specific activity of 40–111 GBq/µmol (1.1–3.0 Ci/µmol). The entire procedure, including the precursor preparation and 18F radiolabeling, takes 7–10 d. PMID:26313478

  4. A rapid, one step molecular identification of Trichoderma citrinoviride and Trichoderma reesei.

    Science.gov (United States)

    Saroj, Dina B; Dengeti, Shrinivas N; Aher, Supriya; Gupta, Anil K

    2015-06-01

    Trichoderma species are widely used as production hosts for industrial enzymes. Identification of Trichoderma species requires a complex molecular biology based identification involving amplification and sequencing of multiple genes. Industrial laboratories are required to run identification tests repeatedly in cell banking procedures and also to prove absence of production host in the product. Such demands can be fulfilled by a brief method which enables confirmation of strain identity. This communication describes one step identification method for two common Trichoderma species; T. citrinoviride and T. reesei, based on identification of polymorphic region in the nucleotide sequence of translation elongation factor 1 alpha. A unique forward primer and common reverse primer resulted in 153 and 139 bp amplicon for T. citrinoviride and T. reesei, respectively. Simplification was further introduced by using mycelium as template for PCR amplification. Method described in this communication allows rapid, one step identification of two Trichoderma species.

  5. Multicenter Evaluation of a Transcription-Reverse Transcription Concerted Assay for Rapid Detection of Mycobacterium tuberculosis Complex in Clinical Specimens ▿

    Science.gov (United States)

    Drouillon, V.; Delogu, G.; Dettori, G.; Lagrange, P. H.; Benecchi, M.; Houriez, F.; Baroli, K.; Ardito, F.; Torelli, R.; Chezzi, C.; Fadda, G.; Herrmann, J.-L.

    2009-01-01

    A European multicenter study was performed to evaluate the performance of a new method, based on the transcription-reverse transcription concerted reaction (TRC-2), which enabled one-step amplification and real-time detection of the Mycobacterium tuberculosis 16S rRNA target directly in clinical specimens. A total of 633 respiratory and nonrespiratory specimens were tested, and the results were compared with those from smears and cultures. A total of 129 patients (Paris center) were followed up in order to evaluate the clinical performance of TRC-2. By using M. tuberculosis complex strains to inoculate sterile sputa, the detection limit of TRC-2 was found to be 30 to 50 CFU/ml. A total of 548 respiratory specimens and 59 extrapulmonary specimens were assessable. For pulmonary specimens, the sensitivities of TRC-2 and acid-fast smear were 86.8% and 50.4%, respectively (P = 0.002). The specificities were 97.5% and 100%, respectively. For extrapulmonary specimens, the sensitivities of TRC-2 and acid-fast smear were 83.3% and 8.3% (P < 0.0001), and the specificities were 95.8% and 100%, respectively. Fifteen of 129 patients were diagnosed with pulmonary tuberculosis (TB). The sensitivities of culture and TRC-2 were 80% (12/15) and 86.7% (13/15) (P = 0.16), and the specificities were 100% and 93.9%, respectively. Based on an 11.6% incidence of TB in our population, the positive predictive values of TRC-2 and culture were 81.3% and 100%, respectively, and the negative predictive values were 98.2% and 97.4%, respectively. These results demonstrated that detection of M. tuberculosis complex in clinical specimens by TRC-2 with ready-to-use reagents was an efficient and rapid method for the diagnosis of pulmonary and extrapulmonary TB. PMID:19741080

  6. One-step method for the production of nanofluids

    Energy Technology Data Exchange (ETDEWEB)

    Kostic, Milivoje (Sycamore, IL); Golubovic, Mihajlo (Chicago, IL); Hull, John (Downers Grove, IL); Choi, Stephen U. S. (Naperville, IL)

    2011-08-16

    A one step method and system for producing nanofluids by a nanoparticle-source evaporation and deposition of the evaporant into a base fluid. The base fluid such oil or ethylene glycol is placed in a rotating cylindrical drum having an adjustable heater-boat-evaporator and heat exchanger-cooler apparatus. As the drum rotates, a thin liquid layer is formed on the inside surface of the drum. An insulated heater-boat-evaporator having an evaporant material (nanoparticle-source) placed within its boat evaporator is adjustably positioned near a portion of the rotating thin liquid layer, the evaporant material being heated thereby evaporating a portion of the evaporant material and forming nanoparticles, the nanoparticles absorbed by the liquid film to form nanofluid.

  7. One-step method for the production of nanofluids

    Energy Technology Data Exchange (ETDEWEB)

    Kostic, Milivoje (Chicago, IL); Golubovic, Mihajlo (Chicago, IL); Hull, John R. (Downers Grove, IL); Choi, Stephen U. S. (Napersville, IL)

    2010-05-18

    A one step method and system for producing nanofluids by a particle-source evaporation and deposition of the evaporant into a base fluid. The base fluid such (i.e. ethylene glycol) is placed in a rotating cylindrical drum having an adjustable heater-boat-evaporator and heat exchanger-cooler apparatus. As the drum rotates, a thin liquid layer is formed on the inside surface of the drum. A heater-boat-evaporator having an evaporant material (particle-source) placed within its boat evaporator is adjustably positioned near a portion of the rotating thin liquid layer, the evaporant material being heated thereby evaporating a portion of the evaporant material, the evaporated material absorbed by the liquid film to form nanofluid.

  8. One-step electrodeposition process to fabricate cathodic superhydrophobic surface

    Energy Technology Data Exchange (ETDEWEB)

    Chen Zhi, E-mail: c2002z@nwpu.edu.cn [Department of Applied Physics, Northwestern Polytechnical University, Xi' an 710129 (China); Li Feng [Department of Applied Physics, Northwestern Polytechnical University, Xi' an 710129 (China); Hao Limei [Department of Applied Physics, Xi' an University of Science and Technology, Xi' an 710054 (China); Chen Anqi; Kong Youchao [Department of Applied Physics, Northwestern Polytechnical University, Xi' an 710129 (China)

    2011-12-01

    In this work, a rapid one-step process is developed to fabricate superhydrophobic cathodic surface by electrodepositing copper plate in an electrolyte solution containing manganese chloride (MnCl{sub 2}{center_dot}4H{sub 2}O), myristic acid (CH{sub 3}(CH{sub 2}){sub 12}COOH) and ethanol. The superhydrophobic surfaces were characterized by means of scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). The shortest electrolysis time for fabricating a superhydrophobic surface is about 1 min, the measured maximum contact angle is 163 Degree-Sign and rolling angle is less than 3 Degree-Sign . Furthermore, this method can be easily extended to other conductive materials. The approach is time-saving and cheap, and it is supposed to have a promising future in industrial fields.

  9. One-step electrodeposition process to fabricate cathodic superhydrophobic surface

    Science.gov (United States)

    Chen, Zhi; Li, Feng; Hao, Limei; Chen, Anqi; Kong, Youchao

    2011-12-01

    In this work, a rapid one-step process is developed to fabricate superhydrophobic cathodic surface by electrodepositing copper plate in an electrolyte solution containing manganese chloride (MnCl2·4H2O), myristic acid (CH3(CH2)12COOH) and ethanol. The superhydrophobic surfaces were characterized by means of scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). The shortest electrolysis time for fabricating a superhydrophobic surface is about 1 min, the measured maximum contact angle is 163° and rolling angle is less than 3°. Furthermore, this method can be easily extended to other conductive materials. The approach is time-saving and cheap, and it is supposed to have a promising future in industrial fields.

  10. An isothermal, label-free, and rapid one-step RNA amplification/detection assay for diagnosis of respiratory viral infections.

    Science.gov (United States)

    Koo, Bonhan; Jin, Choong Eun; Lee, Tae Yoon; Lee, Jeong Hoon; Park, Mi Kyoung; Sung, Heungsup; Park, Se Yoon; Lee, Hyun Jung; Kim, Sun Mi; Kim, Ji Yeun; Kim, Sung-Han; Shin, Yong

    2017-04-15

    Recently, RNA viral infections caused by respiratory viruses, such as influenza, parainfluenza, respiratory syncytial virus, coronavirus, and Middle East respiratory syndrome-coronavirus (MERS-CoV), and Zika virus, are a major public health threats in the world. Although myriads of diagnostic methods based on RNA amplification have been developed in the last decades, they continue to lack speed, sensitivity, and specificity for clinical use. A rapid and accurate diagnostic method is needed for appropriate control, including isolation and treatment of the patients. Here, we report an isothermal, label-free, one-step RNA amplification and detection system, termed as iROAD, for the diagnosis of respiratory diseases. It couples a one-step isothermal RNA amplification method and a bio-optical sensor for simultaneous viral RNA amplification/detection in a label-free and real-time manner. The iROAD assay offers a one-step viral RNA amplification/detection example to rapid analysis (<20min). The detection limit of iROAD assay was found to be 10-times more sensitive than that of real-time reverse transcription-PCR method. We confirmed the clinical utility of the iROAD assay by detecting viral RNAs obtained from 63 human respiratory samples. We envision that the iROAD assay will be useful and potentially adaptable for better diagnosis of emerging infectious diseases including respiratory diseases.

  11. Reverse transcription using random pentadecamer primers increases yield and quality of resulting cDNA

    DEFF Research Database (Denmark)

    Stangegaard, Michael; Dufva, I.H.; Dufva, Hans Martin

    2006-01-01

    oligonucleotides (pentadecamers) consistently, yielded at least 2 fold as much cDNA as did random hexamers using either-poly(A) RNA or an amplified version of messenger RNA (aRNA) as a template. The cDNA generated using pentadecamers did not differ in size distribution or the amount of incorporated label compared...... with cDNA generated with random hexamers. The increased efficiency of priming using random pentadecamers resulted in reverse transcription of > 80% of the template aRNA, while random hexamers induced reverse transcription of only 40% of the template aRNA. This suggests a better coverage...... that random pentadecamers can replace random hexamers in reverse transcription reactions on both poly(A) RNA and amplified RNA, resulting in higher cDNA yields and quality....

  12. The specificity and flexibility of l1 reverse transcription priming at imperfect T-tracts.

    Directory of Open Access Journals (Sweden)

    Clément Monot

    2013-05-01

    Full Text Available L1 retrotransposons have a prominent role in reshaping mammalian genomes. To replicate, the L1 ribonucleoprotein particle (RNP first uses its endonuclease (EN to nick the genomic DNA. The newly generated DNA end is subsequently used as a primer to initiate reverse transcription within the L1 RNA poly(A tail, a process known as target-primed reverse transcription (TPRT. Prior studies demonstrated that most L1 insertions occur into sequences related to the L1 EN consensus sequence (degenerate 5'-TTTT/A-3' sites and frequently preceded by imperfect T-tracts. However, it is currently unclear whether--and to which degree--the liberated 3'-hydroxyl extremity on the genomic DNA needs to be accessible and complementary to the poly(A tail of the L1 RNA for efficient priming of reverse transcription. Here, we employed a direct assay for the initiation of L1 reverse transcription to define the molecular rules that guide this process. First, efficient priming is detected with as few as 4 matching nucleotides at the primer 3' end. Second, L1 RNP can tolerate terminal mismatches if they are compensated within the 10 last bases of the primer by an increased number of matching nucleotides. All terminal mismatches are not equally detrimental to DNA extension, a C being extended at higher levels than an A or a G. Third, efficient priming in the context of duplex DNA requires a 3' overhang. This suggests the possible existence of additional DNA processing steps, which generate a single-stranded 3' end to allow L1 reverse transcription. Based on these data we propose that the specificity of L1 reverse transcription initiation contributes, together with the specificity of the initial EN cleavage, to the distribution of new L1 insertions within the human genome.

  13. Complementary assays reveal a relationship between HIV-1 uncoating and reverse transcription.

    Science.gov (United States)

    Hulme, Amy E; Perez, Omar; Hope, Thomas J

    2011-06-14

    During the early stages of HIV-1 replication the conical capsid composed of p24(CA) protein dissociates from the rest of the cytoplasmic viral complex by a process called uncoating. Although proper uncoating is known to be required for HIV-1 infection, many questions remain about the timing and factors involved in the process. Here we have used two complementary assays to study the process of uncoating in HIV-1-infected cells, specifically looking at the timing of uncoating and its relationship to reverse transcription. We developed a fluorescent microscopy-based uncoating assay that detects the association of p24(CA) with HIV-1 viral complexes in cells. We also used an owl monkey kidney (OMK) cell assay that is based on timed TRIM-CypA-mediated restriction of HIV-1 replication. Results from both assays indicate that uncoating is initiated within 1 h of viral fusion. In addition, treatment with the reverse transcriptase inhibitor nevirapine delayed uncoating in both assays. Analysis of reverse transcription products in OMK cells revealed that the generation of early reverse transcription products coincides with the timing of uncoating in these assays. Collectively, these results suggest that some aspect of reverse transcription has the ability to influence the kinetics of uncoating.

  14. One step facile synthesis of ferromagnetic magnetite nanoparticles

    Science.gov (United States)

    Suppiah, Durga Devi; Abd Hamid, Sharifah Bee

    2016-09-01

    The ferromagnetic properties of magnetite (Fe3O4) were influenced by the nanoparticle size, hence importance were given to the synthesis method. This paper clearly shows that magnetite nanoparticles were successfully synthesized by employing one step controlled precipitation method using a single salt (Iron(II) sulfate) iron precursor. The acquired titration curve from this method provides vital information on the possible reaction mechanism leading to the magnetite (Fe3O4) nanoparticles formation. Goethite (α-FeOOH) was obtained at pH 4, while the continuous addition of hydroxyl ions (OH-) forms iron hydroxides (Fe(OH)2). This subsequently reacts with the goethite, producing magnetite (Fe3O4) at pH 10. Spectroscopy studies validate the magnetite phase existence while structural and morphology analysis illustrates cubic shaped magnetite with an average size of 35 nm was obtained. The synthesized magnetite might be superparamagnetic though lower saturation magnetization (67.5 emu/g) measured at room temperature as compared to bulk magnetite. However the nanoparticles surface anisotropy leads to higher remanence (12 emu/g) and coercivity (117.7 G) making the synthesized magnetite an excellent candidate to be utilized in recording devices. The understanding of the magnetite synthesis mechanism can not only be used to achieve even smaller magnetite nanoparticles but also to prepare different types of iron oxides hydroxides using different iron precursor source.

  15. Methanobactin-mediated one-step synthesis of gold nanoparticles.

    Science.gov (United States)

    Xin, Jia-ying; Cheng, Dan-dan; Zhang, Lan-xuan; Lin, Kai; Fan, Hong-chen; Wang, Yan; Xia, Chun-gu

    2013-11-01

    Preparation of gold nanoparticles with a narrow size distribution has enormous importance in nanotechnology. Methanobactin (Mb) is a copper-binding small peptide that appears to function as an agent for copper sequestration and uptake in methanotrophs. Mb can also bind and catalytically reduce Au (III) to Au (0). In this study, we demonstrate a facile Mb-mediated one-step synthetic route to prepare monodispersed gold nanoparticles. Continuous reduction of Au (III) by Mb can be achieved by using hydroquinone as the reducing agent. The gold nanoparticles have been characterized by UV-visible spectroscopy. The formation and the surface plasmon resonance properties of the gold nanoparticles are highly dependent on the ratio of Au (III) to Mb in solution. X-ray photoelectron spectroscopy (XPS), fluorescence spectra and Fourier transform-infrared spectroscopy (FT-IR) spectra suggest that Mb molecules catalytically reduce Au (III) to Au (0) with the concomitant production of gold nanoparticles, and then, Mb statically adsorbed onto the surface of gold nanoparticles to form an Mb-gold nanoparticles assembly. This avoids secondary nucleation. The formed gold nanoparticles have been demonstrated to be monodispersed and uniform by transmission electron microscopy (TEM) images. Analysis of these particles shows an average size of 14.9 nm with a standard deviation of 1.1 nm. The gold nanoparticles are extremely stable and can resist aggregation, even after several months.

  16. One-step electrospinning of cross-linked chitosan fibers.

    Science.gov (United States)

    Schiffman, Jessica D; Schauer, Caroline L

    2007-09-01

    Chitin is a nitrogen-rich polysaccharide that is abundant in crustaceans, mollusks, insects, and fungi and is the second most abundant organic material found in nature next to cellulose. Chitosan, the N-deacetylated derivative of chitin, is environmentally friendly, nontoxic, biodegradable, and antibacterial. Fibrous mats are typically used in industries for filter media, catalysis, and sensors. Decreasing fiber diameters within these mats causes many beneficial effects such as increased specific surface area to volume ratios. When the intrinsically beneficial effects of chitosan are combined with the enhanced properties of nanofibrous mats, applications arise in a wide range of fields, including medical, packaging, agricultural, and automotive. This is particularly important as innovative technologies that focus around bio-based materials are currently of high urgency, as they can decrease dependencies on fossil fuels. We have demonstrated that Schiff base cross-linked chitosan fibrous mats can be produced utilizing a one-step electrospinning process that is 25 times faster and, therefore, more economical than a previously reported two-step vapor-cross-linking method. These fibrous mats are insoluble in acidic, basic, and aqueous solutions for 72 h. Additionally, this improved production method results in a decreased average fiber diameter, which measures 128 +/- 40 nm. Chemical and structural analyses were conducted utilizing Fourier transform infrared spectroscopy, solubility studies, and scanning electron microscopy.

  17. Methanobactin-Mediated One-Step Synthesis of Gold Nanoparticles

    Directory of Open Access Journals (Sweden)

    Yan Wang

    2013-11-01

    Full Text Available Preparation of gold nanoparticles with a narrow size distribution has enormous importance in nanotechnology. Methanobactin (Mb is a copper-binding small peptide that appears to function as an agent for copper sequestration and uptake in methanotrophs. Mb can also bind and catalytically reduce Au (III to Au (0. In this study, we demonstrate a facile Mb-mediated one-step synthetic route to prepare monodispersed gold nanoparticles. Continuous reduction of Au (III by Mb can be achieved by using hydroquinone as the reducing agent. The gold nanoparticles have been characterized by UV-visible spectroscopy. The formation and the surface plasmon resonance properties of the gold nanoparticles are highly dependent on the ratio of Au (III to Mb in solution. X-ray photoelectron spectroscopy (XPS, fluorescence spectra and Fourier transform-infrared spectroscopy (FT-IR spectra suggest that Mb molecules catalytically reduce Au (III to Au (0 with the concomitant production of gold nanoparticles, and then, Mb statically adsorbed onto the surface of gold nanoparticles to form an Mb-gold nanoparticles assembly. This avoids secondary nucleation. The formed gold nanoparticles have been demonstrated to be monodispersed and uniform by transmission electron microscopy (TEM images. Analysis of these particles shows an average size of 14.9 nm with a standard deviation of 1.1 nm. The gold nanoparticles are extremely stable and can resist aggregation, even after several months.

  18. One Step Hydrogen Generation Through Sorption Enhanced Reforming

    Energy Technology Data Exchange (ETDEWEB)

    Mays, Jeff [Gas Technology Inst., Des Plaines, IL (United States)

    2017-08-03

    One-step hydrogen generation, using Sorption Enhanced Reforming (SER) technology, is an innovative means of providing critical energy and environmental improvements to US manufacturing processes. The Gas Technology Institute (GTI) is developing a Compact Hydrogen Generator (CHG) process, based on SER technology, which successfully integrates previously independent process steps, achieves superior energy efficiency by lowering reaction temperatures, and provides pathways to doubling energy productivity with less environmental pollution. GTI’s prior CHG process development efforts have culminated in an operational pilot plant. During the initial pilot testing, GTI identified two operating risks- 1) catalyst coating with calcium aluminate compounds, 2) limited solids handling of the sorbent. Under this contract GTI evaluated alternative materials (one catalyst and two sorbents) to mitigate both risks. The alternate catalyst met performance targets and did not experience coating with calcium aluminate compounds of any kind. The alternate sorbent materials demonstrated viable operation, with one material enabling a three-fold increase in sorbent flow. The testing also demonstrated operation at 90% of its rated capacity. Lastly, a carbon dioxide co-production study was performed to assess the advantage of the solid phase separation of carbon dioxide- inherent in the CHG process. Approximately 70% lower capital cost is achievable compared to SMR-based hydrogen production with CO2 capture, as well as improved operating costs.

  19. Visual detection of the human metapneumovirus using reverse transcription loop-mediated isothermal amplification with hydroxynaphthol blue dye

    Directory of Open Access Journals (Sweden)

    Wang Xiang

    2012-07-01

    Full Text Available Abstract Background Human metapneumovirus (hMPV is a major cause of acute respiratory infections ranging from wheezing to bronchiolitis and pneumonia in children worldwide. The objective of this study is to develop a visual reverse transcription loop-mediated isothermal amplification (RT-LAMP assay for the detection of hMPV and applied to the clinical samples. Results In this study, visual RT-LAMP assay for hMPV was performed in one step with the addition of hydroxynaphthol blue (HNB, and were used to detect respiratory samples. Six primers, including two outer primers (F3 and B3, two inner primers (FIP, BIP and two loop primers (LF and LB, were designed for hMPV N gene by the online software. Moreover, the RT-LAMP assay showed good specificity and no cross-reactivity was observed with human rhinovirus (HRV, human respiratory syncytial Virus (RSV, or influenza virus A/PR/8/34 (H1N1. The detection limit of the RT-LAMP assay was approximately ten viral RNA copies, lower than that of traditional reverse transcriptase polymerase chain reaction (RT-PCR 100 RNA copies. In the 176 nasopharyngeal samples, 23 (13.1% were conformed as hMPV positive by RT-LAMP, but 18 (10.2% positive by RT-PCR. Conclusion Compared with conventional RT-PCR, the visual hMPV RT-LAMP assay performed well in the aspect of detect time, sensitivity, specificity and visibility. It is anticipated that the RT-LAMP will be used for clinical tests in hospital or field testing during outbreaks and in emergency.

  20. One-step continuous synthesis of functionalized magnetite nanoflowers

    Science.gov (United States)

    Thomas, G.; Demoisson, F.; Chassagnon, R.; Popova, E.; Millot, N.

    2016-04-01

    For the first time, functionalized magnetite nanoparticles (Fe3O4 NPs) that form aggregates with a nanoflower morphology were synthesized using a rapid (11 s) one-step continuous hydrothermal process, which was recently modified, and their application as a T 2 magnetic resonance imaging (MRI) contrast agent was evaluated. The nanoparticles functionalized with 3,4-dihydroxy-L-phenylalanine (LDOPA) or 3,4-dihydroxyhydrocinnamic acid (DHCA) consisted of small crystallites of approximately 15 nm of diameter that assembled to form flower-shaped aggregate structures. The Fe3O4-LDOPA nanoflowers exhibited a high transverse relaxivity, r 2 of 418 ± 10 l mmolFe -1 s-1 at 3 T owing to magnetic dipolar interactions, which is twice as that of the commercial Feridex®/Endorem®. The prepared nanostructures were compared with bare Fe3O4 NPs and citrated Fe3O4 NPs. DHCA, LDOPA, and citric acid (CA) were found to have an anti-oxidizing effect and to influence the crystallite size and the lattice parameter of the NPs. DHCA and LDOPA increased the crystallite size, whereas CA decreased it. Surface modification increased the colloidal stability of NPs as compared to bare NPs. Nanoflower suspensions of Fe3O4-LDOPA NPs were found to be stable in the phosphate-buffered saline, saline medium, and minimal essential medium and formed aggregates of sizes smaller than 120 nm. All samples were found to be superparamagnetic in nature and the highest saturation magnetization was obtained for the Fe3O4-LDOPA samples. These NPs can bind to polymers such as PEG, and to fluorescent and chelating agents owing to the presence of free -NH2 or -COOH groups on the surface of NPs, allowing their use in dual imaging applications.

  1. A reverse transcription loop-mediated isothermal amplification (LAMP) assay for the detection of feline Coronavirus

    National Research Council Canada - National Science Library

    Angelica Stranieri; Stefania Lauzi; Alessia Giordano; Saverio Paltrinieri

    2016-01-01

    ...). The addition of two loop primers allows the reaction time to be of one hour only (Nagamine et al., 2002). The aim of this study was to develop a reverse transcription LAMP assay for an easy and inexpensive detection of feline Coronavirus...

  2. Sequence-specific inhibition of duck hepatitis B virus reverse transcription by peptide nucleic acids (PNA)

    DEFF Research Database (Denmark)

    Robaczewska, Magdalena; Narayan, Ramamurthy; Seigneres, Beatrice

    2005-01-01

    BACKGROUND/AIMS: Peptide nucleic acids (PNAs) appear as promising new antisense agents, that have not yet been examined as hepatitis B virus (HBV) inhibitors. Our aim was to study the ability of PNAs targeting the duck HBV (DHBV) encapsidation signal epsilon to inhibit reverse transcription (RT...

  3. Reversible Histone Acetylation Involved in Transcriptional Regulation of WT1 Gene

    Institute of Scientific and Technical Information of China (English)

    Yangguang SHAO; Jun LU; Cao CHENG; Liguo CUI; Guoping ZHANG; Baiqu HUANG

    2007-01-01

    To validate the involvement of reversible histone acetylation in the transcriptional regulation of human Wilms' tumor 1 gene (WT1), we analyzed the roles of histone deacetylases (HDACs) and histone acetyltransferase in this epigenetic process. Of the six HDACs (HDAC1-6) examined, HDAC4 and HDAC5 were found to have significant repressing effects on the activity of the WT1 reporter gene, as revealed by luciferase reporter assays and quantitative real-time reverse transcription-polymerase chain reaction assays.Luciferase reporter assays showed that the histone acetyltransferase p300 was able to counteract the HDAC4/HDAC5-mediated repression and that p300/CBP synergized with transcription factors Sp1, c-Myb, and Ets-1 in activation of the WT1 reporter. Chromatin immunoprecipitation experiments showed that p300 promotes the acetylation level of histone H3 at the WT1 intronic enhancer. Based on these data, we proposed a hypothetical model for the involvement of reversible histone acetylation in transcriptional regulation of the WT1 gene. This study provides further insight into the mechanisms of transcriptional regulation of the WT1 gene and WT1-associated diseases treatment.

  4. Application of Reverse Transcription-PCR and Real-Time PCR in Nanotoxicity Research

    Science.gov (United States)

    Mo, Yiqun; Wan, Rong; Zhang, Qunwei

    2016-01-01

    Reverse transcription-polymerase chain reaction (RT-PCR) is a relatively simple and inexpensive technique to determine the expression level of target genes and is widely used in biomedical science research including nanotoxicology studies for semiquantitative analysis. Real-time PCR allows for the detection of PCR amplification in the exponential growth phase of the reaction and is much more quantitative than traditional RT-PCR. Although a number of kits and reagents for RT-PCR and real-time PCR are commercially available, the basic principles are the same. Here, we describe the procedures for total RNA isolation by using TRI Reagent, for reverse transcription (RT) by M-MLV reverse transcriptase, and for PCR by GoTaq® DNA Polymerase. And real-time PCR will be performed on an iQ5 multicolor real-time PCR detection system by using iQ™ SYBR Green Supermix. PMID:22975959

  5. Gene expression analysis using strains constructed by NHEJ-mediated one-step promoter cloning in the yeast Kluyveromyces marxianus.

    Science.gov (United States)

    Suzuki, Ayako; Fujii, Hiroshi; Hoshida, Hisashi; Akada, Rinji

    2015-09-01

    Gene expression analysis provides valuable information to evaluate cellular state. Unlike quantitative mRNA analysis techniques like reverse-transcription PCR and microarray, expression analysis using a reporter gene has not been commonly used for multiple-gene analysis, probably due to the difficulty in preparing multiple reporter-gene constructs. To circumvent this problem, we developed a novel one-step reporter-gene construction system mediated by non-homologous end joining (NHEJ) in the yeast Kluyveromyces marxianus. As a selectable reporter gene, the ScURA3 selection marker was fused in frame with a red fluorescent gene yEmRFP (ScURA3:yEmRFP). The N-terminally truncated ScURA3:yEmRFP fragment was prepared by PCR. Promoter sequences were also prepared by PCR using primers containing the sequence of the deleted ScURA3 N-terminus to attach at their 3(') ends. The two DNA fragments were used for the transformation of a ura3(-) strain of K. marxianus, in which two DNA fragments are randomly joined and integrated into the chromosome through NHEJ. Only the correctly aligned fragments produced transformants on uracil-deficient medium and expressed red fluorescence under the control of the introduced promoters. A total of 36 gene promoters involved in glycolysis and other pathways were analyzed. Fluorescence measurements of these strains allowed real-time gene expression analysis in different culture conditions.

  6. One-step multiplex quantitative RT-PCR for the simultaneous detection of viroids and phytoplasmas of pome fruit trees.

    Science.gov (United States)

    Malandraki, Ioanna; Varveri, Christina; Olmos, Antonio; Vassilakos, Nikon

    2015-03-01

    A one-step multiplex real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) based on TaqMan chemistry was developed for the simultaneous detection of Pear blister canker viroid and Apple scar skin viroid along with universal detection of phytoplasmas, in pome trees. Total nucleic acids (TNAs) extraction was performed according to a modified CTAB protocol. Primers and TaqMan MGB probes for specific detection of the two viroids were designed in this study, whereas for phytoplasma detection published universal primers and probe were used, with the difference that the later was modified to carry a MGB quencher. The pathogens were detected simultaneously in 10-fold serial dilutions of TNAs from infected plant material into TNAs of healthy plant up to dilutions 10(-5) for viroids and 10(-4) for phytoplasmas. The multiplex real-time assay was at least 10 times more sensitive than conventional protocols for viroid and phytoplasma detection. Simultaneous detection of the three targets was achieved in composite samples at least up to a ratio of 1:100 triple-infected to healthy tissue, demonstrating that the developed assay has the potential to be used for rapid and massive screening of viroids and phytoplasmas of pome fruit trees in the frame of certification schemes and surveys.

  7. Exogenous reference gene normalization for real-time reverse transcription-polymerase chain reaction analysis under dynamic endogenous transcription

    Institute of Scientific and Technical Information of China (English)

    Stephen Johnston; Zachary Gallaher; Krzysztof Czaja

    2012-01-01

    Quantitative real-time reverse transcription-polymerase chain reaction (qPCR) is widely used to investigate transcriptional changes following experimental manipulations to the nervous system. Despite the widespread utilization of qPCR, the interpretation of results is marred by the lack of a suitable reference gene due to the dynamic nature of endogenous transcription. To address this inherent deficiency, we investigated the use of an exogenous spike-in mRNA, luciferase, as an internal reference gene for the 2-ΔΔCt normalization method. To induce dynamic transcription, we systemically administered capsaicin, a neurotoxin selective for C-type sensory neurons expressing the TRPV-1 receptor, to adult male Sprague-Dawley rats. We later isolated nodose ganglia for qPCR analysis with the reference being either exogenous luciferase mRNA or the commonly used endogenous reference β-III tubulin. The exogenous luciferase mRNA reference clearly demonstrated the dynamic expression of the endogenous reference. Furthermore, variability of the endogenous reference would lead to misinterpretation of other genes of interest. In conclusion, traditional reference genes are often unstable under physiologically normal situations, and certainly unstable following the damage to the nervous system. The use of exogenous spike-in reference provides a consistent and easily implemented alternative for the analysis of qPCR data.

  8. Virion-incorporated alpha-enolase suppresses the early stage of HIV-1 reverse transcription.

    Science.gov (United States)

    Kishimoto, Naoki; Iga, Nozomi; Yamamoto, Kengo; Takamune, Nobutoki; Misumi, Shogo

    2017-03-04

    Human immunodeficiency virus type-1 (HIV-1) particles contain not only viral-encoded but also host-encoded proteins. Interestingly, several studies showed that host proteins play a critical role in viral infectivity, replication and/or immunoreactivity in the next target cells. Here, we show that alpha-enolase (ENO1) is incorporated into HIV-1 virions and the virion-incorporated ENO1 prevents the early stage of HIV-1 reverse transcription. We found that viral particles contain two isoforms of ENO1 with different isoelectric points by two-dimensional electrophoresis. Suppression of ENO1 expression by RNA interference in the HIV-1 producer cells decreased ENO1 incorporation into virions without altering the packaging of viral structural proteins and viral production but increased viral infectivity. Although the low-level-ENO1-packaging virus maintained comparable levels of reverse transcriptase activity, viral genomic RNA and tRNA(Lys3) packaging to the control virus, its levels of early cDNA products of reverse transcription were higher than those of the control virus. In contrast, the high-level-ENO1-packaging virus, which was produced from ENO1-overexpressing cells, showed decreased infectivity and the levels of early cDNA products. Taken together, these findings reveal a novel function of ENO1 as a negative regulation factor targeting HIV-1 reverse transcription.

  9. Development of a neutralization assay for influenza virus using an endpoint assessment based on quantitative reverse-transcription PCR.

    Directory of Open Access Journals (Sweden)

    Belete Teferedegne

    Full Text Available A microneutralization assay using an ELISA-based endpoint assessment (ELISA-MN is widely used to measure the serological response to influenza virus infection and vaccination. We have developed an alternative microneutralization assay for influenza virus using a quantitative reverse transcription PCR-based endpoint assessment (qPCR-MN in order to improve upon technical limitations associated with ELISA-MN. For qPCR-MN, infected MDCK-London cells in 96-well cell-culture plates are processed with minimal steps such that resulting samples are amenable to high-throughput analysis by downstream one-step quantitative reverse transcription PCR (qRT-PCR; SYBR Green chemistry with primers targeting a conserved region of the M1 gene of influenza A viruses. The growth curves of three recent vaccine strains demonstrated that the qRT-PCR signal detected at 6 hours post-infection reflected an amplification of at least 100-fold over input. Using ferret antisera, we have established the feasibility of measuring virus neutralization at 6 hours post-infection, a duration likely confined to a single virus-replication cycle. The neutralization titer for qPCR-MN was defined as the highest reciprocal serum dilution necessary to achieve a 90% inhibition of the qRT-PCR signal; this endpoint was found to be in agreement with ELISA-MN using the same critical reagents in each assay. qPCR-MN was robust with respect to assay duration (6 hours vs. 12 hours. In addition, qPCR-MN appeared to be compliant with the Percentage Law (i.e., virus neutralization results appear to be consistent over an input virus dose ranging from 500 to 12,000 TCID(50. Compared with ELISA-MN, qPCR-MN might have inherent properties conducive to reducing intra- and inter-laboratory variability while affording suitability for automation and high-throughput uses. Finally, our qRT-PCR-based approach may be broadly applicable to the development of neutralization assays for a wide variety of viruses.

  10. [Rapid detection of Macrobrachium rosenbergii nodavirus isolated in China by a reverse-transcription loop-mediated isothermal amplification assay combined with a lateral flow dipstick method].

    Science.gov (United States)

    Lin, Feng; Liu, Li; Hao, Gui-Jie; Cao, Zheng; Sheng, Peng-Cheng; Wu, Ying-Lei; Shen, Jin-Yu

    2014-09-01

    White coloration of the muscle of the giant river prawn (Macrobrachium rosenbergii) is a serious problem in China. The Macrobrachium rosenbergii Nodavirus (MrNV) has been confirmed to be the pathogen that causes this disorder. To develop a rapid, sensitive and specific technology for the detection of Macrobrachium rosenbergii Nodavirus isolated from China (MrNV-China), a reverse-transcription loop- mediated isothermal amplification assay combined with a lateral flow dipstick (RT-LAMP-LFD) assay method is described. A set of four primers and a labeled probe were designed specifically to recognize six distinct regions of the MrNV RNA2 gene. Results showed the sensitivity of the RT-LAMP-LFD assay was ten-times higher than the reverse-transcription loop-mediated isothermal amplification assay (RT-LAMP) with agarose gel electrophoresis. The assay was conducted with one-step amplification at 61°C in a single tube within 45 min. No product was generated from shrimps infected with other viruses, including DNA viruses (infectious hypodermal and hematopoietic necrosis virus (IHHNV); white spot syndrome virus (WSSV)) and RNA viruses (Taura syndrome virus (TSV); infectious myonecrosis virus (IMNV); yellow head virus (YHV)). Results were visualized by the LFD method. Therefore, the described rapid and sensitive assay is potentially useful for MrNV detection.

  11. Development and evaluation of a simple assay for Marburg virus detection using a reverse transcription-loop-mediated isothermal amplification method.

    Science.gov (United States)

    Kurosaki, Yohei; Grolla, Allen; Fukuma, Aiko; Feldmann, Heinz; Yasuda, Jiro

    2010-07-01

    Marburg virus (MARV) causes a severe hemorrhagic fever in humans with a high mortality rate. The rapid and accurate identification of the virus is required to appropriately provide infection control and outbreak management. Here, we developed and evaluated a one-step reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for the rapid and simple detection of MARV. By combining two sets of primers specific for the Musoke and Ravn genetic lineages, a multiple RT-LAMP assay detected MARV strains of both lineages, and no cross-reactivity with other hemorrhagic fever viruses (Ebola virus and Lassa virus) was observed. The assay could detect 10(2) copies of the viral RNA per tube within 40 min by real-time monitoring of the turbidities of the reaction mixtures. The assay was further evaluated using viral RNA extracted from clinical specimens collected in the 2005 Marburg hemorrhagic fever outbreak in Angola and yielded positive results for samples containing MARV at greater than 10(4) 50% tissue culture infective doses/ml, exhibiting 78% (14 of 18 samples positive) consistency with the results of a reverse transcription-PCR assay carried out in the field laboratory. The results obtained by both agarose gel electrophoresis and naked-eye judgment indicated that the RT-LAMP assay developed in this study is an effective tool for the molecular detection of MARV. Furthermore, it seems suitable for use for field diagnostics or in laboratories in areas where MARV is endemic.

  12. Transcript Regulation of Human Telomerase Reverse Transcriptase by c-myc and mad1

    Institute of Scientific and Technical Information of China (English)

    Lin ZOU; Peng-Hui ZHANG; Chun-Li LUO; Zhi-Guang TU

    2005-01-01

    Telomerase activity is highly positive correlated to most malignant neoplasms. Human telomerase reverse transcriptase (hTERT) is the rate-limiting factor of telomerase activity. Recent studies have shown that the expression of hTERT is mainly determined by its transcript regulation. Among the transcript regulation factors of hTERT, c-myc and mad1 are well known. Here, we constructed c-myc and mad1 eukaryotic expression vectors, then co-transfected them with the wild-type (Tw) or mutant hTERT promoter (Td)luciferase reporter plasmid, which were double-mutated in the E-box sequences from CACGTG to CACCTG of Tw. The change of luciferase activity in different cells was detected. The results showed that Tw was obviously activated in T24 and EJ bladder cancer cells, but not in normal fibrocytes. c-myc and mad1 had positive and negative effects respectively on the Tw transcript in a dose-dependent manner, while the roles of c-myc and mad1 in regulating the Td transcript were reversed. c-myc combined with mad1 can downregulate Tw but not Td. These observations indicate that c-myc and mad1 can regulate the hTERT transcript in a different manner in hTERT positive cells, but not in normal cells. This may provide an insight into some telomerase-related carcinogenesis mechanisms.

  13. Reverse Transcription-PCR Analysis of the Regulation of the Manganese Peroxidase Gene Family

    OpenAIRE

    Gettemy, Jessica M.; Ma, Biao; Alic, Margaret; Gold, Michael H.

    1998-01-01

    Manganese peroxidase (MnP) gene expression in the lignin-degrading fungus Phanerochaete chrysosporium is regulated by nutrient nitrogen levels and by Mn(II), the substrate for the enzyme, as well as by heat shock and other factors. Reverse transcription-PCR (RT-PCR) of total RNA can distinguish the mRNAs of each of the three sequenced P. chrysosporium mnp genes, i.e., mnp1, mnp2, and mnp3. Quantitative RT-PCR demonstrates that each of the three transcripts is present at a similar low basal le...

  14. Utility of IgM ELISA, TaqMan real-time PCR, reverse transcription PCR, and RT-LAMP assay for the diagnosis of Chikungunya fever.

    Science.gov (United States)

    Reddy, Vijayalakshmi; Ravi, Vasanthapuram; Desai, Anita; Parida, Manmohan; Powers, Ann M; Johnson, Barbara W

    2012-11-01

    Chikungunya fever a re-emerging infection with expanding geographical boundaries, can mimic symptoms of other infections like dengue, malaria which makes the definitive diagnosis of the infection important. The present study compares the utility of four laboratory diagnostic methods viz. IgM capture ELISA, an in house reverse transcription PCR for the diagnosis of Chikungunya fever, TaqMan real-time PCR, and a one step reverse transcription-loop mediated isothermal amplification assay (RT-LAMP). Out of the 70 serum samples tested, 29 (41%) were positive for Chikungunya IgM antibody by ELISA and 50 (71%) samples were positive by one of the three molecular assays. CHIKV specific nucleic acid was detected in 33/70 (47%) by reverse transcription PCR, 46/70 (66%) by TaqMan real-time PCR, and 43/70 (62%) by RT-LAMP assay. A majority of the samples (62/70; 89%) were positive by at least one of the four assays used in the study. The molecular assays were more sensitive for diagnosis in the early stages of illness (2-5 days post onset) when antibodies were not detectable. In the later stages of illness, the IgM ELISA is a more sensitive diagnostic test. In conclusion we recommend that the IgM ELISA be used as an initial screening test followed one of the molecular assays in samples that are collected in the early phase of illness and negative for CHIKV IgM antibodies. Such as approach would enable rapid confirmation of the diagnosis and implementation of public health measures especially during outbreaks. Copyright © 2012 Wiley Periodicals, Inc.

  15. Evaluation of reverse transcription-PCR protocols based on the fusion gene for diagnosis of bovine respiratory syncytial virus infections

    OpenAIRE

    Selim A.; Gaede W.

    2013-01-01

    Bovine respiratory syncytial virus (BRSV) is a pneumovirus in the family paramyxoviridae, is an important cause of acute respiratory disease in postweaning calves and feedlot cattle. The real-time reverse transcriptase PCR protocols were developed to detect BRSV infection in infected animals. The sensitivity of RT-PCR protocols based on fusion gene were evaluated using different Mastermixes such as Qiagen One Step RT-PCR (Qiagen) for conventional RT-PCR, Su...

  16. Ketamine and Imipramine Reverse Transcriptional Signatures of Susceptibility and Induce Resilience-Specific Gene Expression Profiles.

    Science.gov (United States)

    Bagot, Rosemary C; Cates, Hannah M; Purushothaman, Immanuel; Vialou, Vincent; Heller, Elizabeth A; Yieh, Lynn; LaBonté, Benoit; Peña, Catherine J; Shen, Li; Wittenberg, Gayle M; Nestler, Eric J

    2017-02-15

    Examining transcriptional regulation by antidepressants in key neural circuits implicated in depression and understanding the relation to transcriptional mechanisms of susceptibility and natural resilience may help in the search for new therapeutic agents. Given the heterogeneity of treatment response in human populations, examining both treatment response and nonresponse is critical. We compared the effects of a conventional monoamine-based tricyclic antidepressant, imipramine, and a rapidly acting, non-monoamine-based antidepressant, ketamine, in mice subjected to chronic social defeat stress, a validated depression model, and used RNA sequencing to analyze transcriptional profiles associated with susceptibility, resilience, and antidepressant response and nonresponse in the prefrontal cortex (PFC), nucleus accumbens, hippocampus, and amygdala. We identified similar numbers of responders and nonresponders after ketamine or imipramine treatment. Ketamine induced more expression changes in the hippocampus; imipramine induced more expression changes in the nucleus accumbens and amygdala. Transcriptional profiles in treatment responders were most similar in the PFC. Nonresponse reflected both the lack of response-associated gene expression changes and unique gene regulation. In responders, both drugs reversed susceptibility-associated transcriptional changes and induced resilience-associated transcription in the PFC. We generated a uniquely large resource of gene expression data in four interconnected limbic brain regions implicated in depression and its treatment with imipramine or ketamine. Our analyses highlight the PFC as a key site of common transcriptional regulation by antidepressant drugs and in both reversing susceptibility- and inducing resilience-associated molecular adaptations. In addition, we found region-specific effects of each drug, suggesting both common and unique effects of imipramine versus ketamine. Copyright © 2016 Society of Biological

  17. Rapid detection of newly isolated Tembusu-related Flavivirus by reverse-transcription loop-mediated isothermal amplification assay

    Directory of Open Access Journals (Sweden)

    Wang Youling

    2011-12-01

    Full Text Available Abstract Background From April 2010 to January 2011, a severe new viral disease had devastated most duck-farming regions in China. This disease affected not only laying ducks but also meat ducks, causing huge economic losses for the poultry industry. The objective of this study is to develop a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP assay for the detection of the new virus related to Tembusu-related Flavivirus. Results The RT-LAMP assay is very simple and rapid, and the amplification can be completed within 50 min under isothermal conditions at 63°C by a set of 6 primers targeting the E gene based on the sequences analysis of the newly isolated viruses and other closely related Flavivirus.The monitoring of gene amplification can also be visualized by using SYBR green I fluorescent dye. In addition, the RT-LAMP assay for newly isolated Tembusu-related Flavivirus showed higher sensitivity with an RNA detection-limit of 2 copies/μL compared with 190 copies/μL of the conventional RT-PCR method. The specificity was identified without cross reaction to other common avian pathogens. By screening a panel of clinical samples this method was more feasible in clinical settings and there was higher positive coincidence rate than conventional RT-PCR and virus isolation. Conclusion The RT-LAMP assay for newly isolated Tembusu-related Flavivirus is a valuable tool for the rapid and real-time detection not only in well-equipped laboratories but also in general conditions.

  18. Detection of Babesia microti parasites by highly sensitive 18S rRNA reverse transcription PCR.

    Science.gov (United States)

    Hanron, Amelia E; Billman, Zachary P; Seilie, Annette M; Chang, Ming; Murphy, Sean C

    2017-03-01

    Babesia are increasingly appreciated as a cause of transfusion-transmitted infection. Sensitive methods are needed to screen blood products. We report herein that B. microti 18S rRNA is over 1,000-fold more abundant than its coding genes, making reverse transcription PCR (RT-PCR) much more sensitive than PCR. Babesia 18S rRNA may be useful for screening the blood supply.

  19. Detection of All Species of the Genus Alphavirus by Reverse Transcription-PCR with Diagnostic Sensitivity▿

    OpenAIRE

    Grywna, K.; Kupfer, B.; Panning, M.; Drexler, J. F.; Emmerich, P.; Drosten, C.; Kummerer, B. M.

    2010-01-01

    Clinical arbovirus screening requires exclusion of a broad range of viruses with as few assays as possible. We present a reverse transcription-PCR (RT-PCR) for the detection of all species of the genus Alphavirus qualified for exclusion screening (limit of detection [LOD], 5 to 100 RNA copies per reaction across all Alphavirus species; detection of viremia down to ca. 10,000 copies per ml).

  20. A reverse transcription-polymerase chain reaction assay for the detection of avian pneumovirus (Colorado strain).

    Science.gov (United States)

    Ali, A; Reynolds, D L

    1999-01-01

    A reverse transcription-polymerase chain reaction assay was developed for the detection of avian pneumovirus (Colorado strain) (APV-Col). The specific primers were designed from the published sequence of the matrix protein gene of APV-Col. The primers amplified a product of 631 nucleotides from APV-Col. The assay identified only APV-Col and did not react with Newcastle disease virus and infectious bronchitis virus.

  1. Detection of Human Picornaviruses by Multiplex Reverse Transcription-PCR and Liquid Hybridization

    OpenAIRE

    Jokela, Pia; Joki-Korpela, Päivi; Maaronen, Marita; Glumoff, Virpi; Hyypiä, Timo

    2005-01-01

    A qualitative multiplex reverse transcription (RT)-PCR and liquid hybridization assay for the detection of human enteroviruses, rhinoviruses, parechoviruses, and Aichi virus was developed. Furthermore, a separate assay for the recognition of hepatitis A virus was established to complement the test pattern so that all human picornaviruses were covered. The amplicons, which represented the 5′ untranslated regions of the viral RNA genomes, were identified in liquid hybridization reactions with g...

  2. One-step synthesis of hierarchically porous carbons for high-performance electric double layer supercapacitors

    Science.gov (United States)

    Zhang, Haitao; Zhang, Lei; Chen, Jun; Su, Hai; Liu, Fangyan; Yang, Weiqing

    2016-05-01

    With plenty of unique porous structure at micro-/nano scale, hierarchically porous carbons (HPCs) are promising for usage in advanced electric double layer supercapacitors (EDLCs) as the electrode materials. However, wide-range adoption of HPC for practical application is largely shadowed by its extremely complex synthesis process with considerably low production efficiency. Herein we reported a simple template-free, one-step sintering method, to massively produce the HPCs for high-performance EDLCs. Resorting to the 3D structure modification of the wide pore size distribution, high surface area of HPCs (up to 3000 m2 g-1) was achieved. By using 1 M Na2SO4 as electrolyte, the as-fabricated HPCs based EDLCs can be operated reversibly over a wide voltage window of 1.6 V with superior specific capacitance of 240 F g-1 under a current density of 0.5 A g-1. In the meanwhile, the EDLCs exhibit excellent rate capability (high power density of 16 kW kg-1 at 10.2 Wh kg-1) and long-term cycling stability with 9% loss of its initial capacitance after 2000 cycles. This output performance distinguished itself among most of the carbon-based EDLCs with neutral aqueous electrolyte. Thus, the template-free one-step sintering method produced HPCs for EDLCs represents a new approach for high-performance energy storage.

  3. NFAT5 regulates transcription of the mouse telomerase reverse transcriptase gene

    Energy Technology Data Exchange (ETDEWEB)

    Fujiki, Tsukasa; Udono, Miyako; Kotake, Yojiro; Yamashita, Makiko; Shirahata, Sanetaka; Katakura, Yoshinori, E-mail: katakura.yoshinori.528@m.kyushu-u.ac.jp

    2010-12-10

    We aimed to clarify the transcription-regulation mechanisms of the mouse telomerase reverse transcriptase gene (mTERT). First, we searched for the promoter region required for transcriptional activation of mTERT and identified an enhancer cis-element (named mTERT-EE) located between - 200 and - 179 bp of the mouse TERT gene (mTERT). EMSA results suggested that nuclear factor of activated T cells (NFAT) member proteins bind to mTERT-EE. We then identified NFAT5 as the factor binding to mTERT-EE and found that it activates the transcription of the mTERT core promoter. The results that siRNA directed against NFAT5 significantly reduced mTERT expression and mTERT core promoter activity and that the expressions of NFAT5 and mTERT were well correlated in various mouse tissues except liver suggest that NFAT5 dominantly and directly regulates mTERT expression. To clarify their functionality further, we investigated the effect of hypertonic stress, a known stimulus affecting the expression and transcriptional activity of NFAT5, on mTERT expression. The result indicated that hypertonic stress activates mTERT transcription via the activation and recruitment of NFAT5 to the mTERT promoter. These results provide useful information about the transcription-regulation mechanisms of mTERT.

  4. Development of a highly sensitive real-time one step RT-PCR combined complementary locked primer technology and conjugated minor groove binder probe

    Directory of Open Access Journals (Sweden)

    Lee Sunhwa

    2011-06-01

    Full Text Available Abstract Background Enterovirus (EV infections are commonly associated with encephalitis and meningitis. Detection of enteroviral RNA in clinical specimens has been demonstrated to improve the management of patients, by ruling out other causes of disease. Method To develop a sensitive and reliable assay for routine laboratory diagnosis, we developed a real-time one step reverse transcription polymerase chain reaction (RT-PCR assay with minor groove binder probes and primers modified with complementary locked primer technology (TMC-PCR. We checked the sensitivity of the developed assay by comparing it to a previously published TaqMan probe real-time one-step RT-PCR (TTN-PCR procedure using enteroviral isolates, Enterovirus Proficiency panels from Quality Control on Molecular Diagnostics (QCMD-2007, and clinical specimens from patients with suspected EV infections. Results One hundred clinical specimens from 158 suspected viral meningitis cases were determined to be positive by the TMC-PCR assay (63.29%, whereas only 60 were found to be positive by the TTN-PCR assay (37.97%. The positive and negative agreements between the TMC-PCR and TTN-PCR assays were 100% and 59.2%, respectively. Conclusion This data suggest that the TMC-PCR assay may be suitable for routine diagnostic screening from patient suspected EV infection.

  5. One-step reduced kinetics for lean hydrogen-air deflagration

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez-Galisteo, D.; Sanchez, A.L. [Area de Mecanica de Fluidos, Univ. Carlos III de Madrid, Leganes 28911 (Spain); Linan, A. [ETSI Aeronauticos, Pl. Cardenal Cisneros 3, Madrid 28040 (Spain); Williams, F.A. [Dept. of Mechanical and Aerospace Engineering, University of California San Diego, La Jolla, CA 92093-0411 (United States)

    2009-05-15

    A short mechanism consisting of seven elementary reactions, of which only three are reversible, is shown to provide good predictions of hydrogen-air lean-flame burning velocities. This mechanism is further simplified by noting that over a range of conditions of practical interest, near the lean flammability limit all reaction intermediaries have small concentrations in the important thin reaction zone that controls the hydrogen-air laminar burning velocity and therefore follow a steady state approximation, while the main species react according to the global irreversible reaction 2H{sub 2} + O{sub 2} {yields} 2H{sub 2}O. An explicit expression for the non-Arrhenius rate of this one-step overall reaction for hydrogen oxidation is derived from the seven-step detailed mechanism, for application near the flammability limit. The one-step results are used to calculate flammability limits and burning velocities of planar deflagrations. Furthermore, implications concerning radical profiles in the deflagration and reasons for the success of the approximations are clarified. It is also demonstrated that adding only two irreversible direct recombination steps to the seven-step mechanism accurately reproduces burning velocities of the full detailed mechanism for all equivalence ratios at normal atmospheric conditions and that an eight-step detailed mechanism, constructed from the seven-step mechanism by adding to it the fourth reversible shuffle reaction, improves predictions of O and OH profiles. The new reduced-chemistry descriptions can be useful for both analytical and computational studies of lean hydrogen-air flames, decreasing required computation times. (author)

  6. Targeted HIV-1 Latency Reversal Using CRISPR/Cas9-Derived Transcriptional Activator Systems.

    Directory of Open Access Journals (Sweden)

    Julia K Bialek

    Full Text Available CRISPR/Cas9 technology is currently considered the most advanced tool for targeted genome engineering. Its sequence-dependent specificity has been explored for locus-directed transcriptional modulation. Such modulation, in particular transcriptional activation, has been proposed as key approach to overcome silencing of dormant HIV provirus in latently infected cellular reservoirs. Currently available agents for provirus activation, so-called latency reversing agents (LRAs, act indirectly through cellular pathways to induce viral transcription. However, their clinical performance remains suboptimal, possibly because reservoirs have diverse cellular identities and/or proviral DNA is intractable to the induced pathways. We have explored two CRISPR/Cas9-derived activator systems as targeted approaches to induce dormant HIV-1 proviral DNA. These systems recruit multiple transcriptional activation domains to the HIV 5' long terminal repeat (LTR, for which we have identified an optimal target region within the LTR U3 sequence. Using this target region, we demonstrate transcriptional activation of proviral genomes via the synergistic activation mediator complex in various in culture model systems for HIV latency. Observed levels of induction are comparable or indeed higher than treatment with established LRAs. Importantly, activation is complete, leading to production of infective viral particles. Our data demonstrate that CRISPR/Cas9-derived technologies can be applied to counteract HIV latency and may therefore represent promising novel approaches in the quest for HIV elimination.

  7. Validation of Reference Genes for Transcriptional Analyses in Pleurotus ostreatus by Using Reverse Transcription-Quantitative PCR.

    Science.gov (United States)

    Castanera, Raúl; López-Varas, Leticia; Pisabarro, Antonio G; Ramírez, Lucía

    2015-06-15

    Recently, the lignin-degrading basidiomycete Pleurotus ostreatus has become a widely used model organism for fungal genomic and transcriptomic analyses. The increasing interest in this species has led to an increasing number of studies analyzing the transcriptional regulation of multigene families that encode extracellular enzymes. Reverse transcription (RT) followed by real-time PCR is the most suitable technique for analyzing the expression of gene sets under multiple culture conditions. In this work, we tested the suitability of 13 candidate genes for their use as reference genes in P. ostreatus time course cultures for enzyme production. We applied three different statistical algorithms and obtained a combination of stable reference genes for optimal normalization of RT-quantitative PCR assays. This reference index can be used for future transcriptomic analyses and validation of transcriptome sequencing or microarray data. Moreover, we analyzed the expression patterns of a laccase and a manganese peroxidase (lacc10 and mnp3, respectively) in lignocellulose and glucose-based media using submerged, semisolid, and solid-state fermentation. By testing different normalization strategies, we demonstrate that the use of nonvalidated reference genes as internal controls leads to biased results and misinterpretations of the biological responses underlying expression changes. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  8. How Many Microorganisms Are Present? Quantitative Reverse Transcription PCR (qRT-PCR)

    Science.gov (United States)

    Price, Andy; Álvarez, Laura Acuña; Whitby, Corinne; Larsen, Jan

    Quantitative reverse transcription PCR (qRT-PCR) is a variation of conventional quantitative or real-time PCR, whereby mRNA is first converted into the complementary DNA (cDNA) by reverse transcription, the cDNA is then subsequently quantified by qPCR. The use of mRNA as the initial template allows the quantification of gene transcripts, rather than gene copy numbers. mRNA is only produced by actively metabolising cells and is produced by its corresponding gene to provide a 'blueprint' in order for a cell to manufacture a specific protein. Conventional qPCR detects not only DNA present in actively metabolising cells but also inactive and dead cells. qRT-PCR has the advantage that only actively metabolising cells are detected, hence provides a more reliable measure of microbial activity in oilfield samples. When qRT-PCR is combined with primers and probes for specific genes, the activity of microbial processes important in the oilfield, such as sulphate reduction, methanogenesis and nitrate reduction can be monitored.

  9. Transcription Regulation of the Human Telomerase Reverse Transcriptase (hTERT Gene

    Directory of Open Access Journals (Sweden)

    Muhammad Khairul Ramlee

    2016-08-01

    Full Text Available Embryonic stem cells and induced pluripotent stem cells have the ability to maintain their telomere length via expression of an enzymatic complex called telomerase. Similarly, more than 85%–90% of cancer cells are found to upregulate the expression of telomerase, conferring them with the potential to proliferate indefinitely. Telomerase Reverse Transcriptase (TERT, the catalytic subunit of telomerase holoenzyme, is the rate-limiting factor in reconstituting telomerase activity in vivo. To date, the expression and function of the human Telomerase Reverse Transcriptase (hTERT gene are known to be regulated at various molecular levels (including genetic, mRNA, protein and subcellular localization by a number of diverse factors. Among these means of regulation, transcription modulation is the most important, as evident in its tight regulation in cancer cell survival as well as pluripotent stem cell maintenance and differentiation. Here, we discuss how hTERT gene transcription is regulated, mainly focusing on the contribution of trans-acting factors such as transcription factors and epigenetic modifiers, as well as genetic alterations in hTERT proximal promoter.

  10. Counterselection of prokaryotic ribosomal RNA during reverse transcription using non-random hexameric oligonucleotides.

    Science.gov (United States)

    Gonzalez, J M; Robb, F T

    2007-12-01

    Ribosomal RNA (rRNA) is the major component in total RNA extracts, interfering with the synthesis of cDNA corresponding to messenger RNA (mRNA). In this study, we present a novel strategy for selectively discriminating against rRNA and favoring mRNA from prokaryotes during synthesis of cDNA by reverse transcriptase. Our technique is based on the fact that rRNA sequences, in many species, are G+C rich relative to the genome at large, and highly conserved among prokaryotes. The sequence TTTT is therefore rarely found in rRNA sequences. However, TTTT priming sites are found at a much higher frequency in protein-encoding gene sequences. We designed specific hexamers (HD/DHTTTT) to prime reverse transcription reactions resulting in a selective synthesis of cDNA corresponding to mRNA from prokaryotic total RNA extractions.

  11. Analysis of liver connexin expression using reverse transcription quantitative real-time polymerase chain reaction

    Science.gov (United States)

    Maes, Michaël; Willebrords, Joost; Crespo Yanguas, Sara; Cogliati, Bruno; Vinken, Mathieu

    2016-01-01

    Summary Although connexin production is mainly regulated at the protein level, altered connexin gene expression has been identified as the underlying mechanism of several pathologies. When studying the latter, appropriate methods to quantify connexin mRNA levels are required. The present chapter describes a well-established reverse transcription quantitative real-time polymerase chain reaction procedure optimized for analysis of hepatic connexins. The method includes RNA extraction and subsequent quantification, generation of complementary DNA, quantitative real-time polymerase chain reaction and data analysis. PMID:27207283

  12. Development of one-step SYBR Green real-time RT-PCR for quantifying bovine viral diarrhea virus type-1 and its comparison with conventional RT-PCR

    Directory of Open Access Journals (Sweden)

    Lou Sai

    2011-07-01

    Full Text Available Abstract Background Bovine viral diarrhea virus (BVDV is a worldwide pathogen in cattle and acts as a surrogate model for hepatitis C virus (HCV. One-step real-time fluorogenic quantitative reverse transcription polymerase chain reaction (RT-PCR assay based on SYBR Green I dye has not been established for BVDV detection. This study aims to develop a quantitative one-step RT-PCR assay to detect BVDV type-1 in cell culture. Results One-step quantitative SYBR Green I RT-PCR was developed by amplifying cDNA template from viral RNA and using in vitro transcribed BVDV RNA to establish a standard curve. The assay had a detection limit as low as 100 copies/ml of BVDV RNA, a reaction efficiency of 103.2%, a correlation coefficient (R2 of 0.995, and a maximum intra-assay CV of 2.63%. It was 10-fold more sensitive than conventional RT-PCR and can quantitatively detect BVDV RNA levels from 10-fold serial dilutions of titrated viruses containing a titer from 10-1 to 10-5 TCID50, without non-specific amplification. Melting curve analysis showed no primer-dimers and non-specific products. Conclusions The one-step SYBR Green I RT-PCR is specific, sensitive and reproducible for the quantification of BVDV in cell culture. This one-step SYBR Green I RT-PCR strategy may be further optimized as a reliable assay for diagnosing and monitoring BVDV infection in animals. It may also be applied to evaluate candidate agents against HCV using BVDV cell culture model.

  13. Rapid detection and differentiation of wild-type and three attenuated lapinized vaccine strains of classical swine fever virus by reverse transcription polymerase chain reaction.

    Science.gov (United States)

    Pan, Chu-Hsiang; Jong, Ming-Hwa; Huang, Yu-Liang; Huang, Tien-Shine; Chao, Parn-Hwa; Lai, Shiow-Suey

    2008-07-01

    A simple one-step reverse transcription polymerase chain reaction (RT-PCR) method was developed based on T-rich insertions in the viral genome for simultaneous detection and differentiation of wild type and vaccine strains of Classical swine fever virus (CSFV). The CSFV-specific primers were designed to contain the sequences of the T-rich insertion sites that exist uniquely in the 3' nontranslated regions (3' NTR) of the genome of lapinized CSFV vaccine strains. By using a one-step RT-PCR or a nested PCR followed by an agarose gel electrophoresis or a multicapillary electrophoresis, the wild-type and lapinized vaccine strains of CSFV in clinical samples could be detected and accurately distinguished. These assays can be applied to at least 3 attenuated lapinized vaccine strains, lapinized Philippines Coronel (LPC), hog cholera lapinized virus (HCLV), and Chinese strain (C strain). The detection limit of the wild-type virus was 6.3 TCID(50) (50% tissue culture infective dose)/ml for RT-PCR and 0.63 TCID(50)/ml for nested PCR. In previous studies, notable T-rich insertions of 12-13 nucleotides (nt) were found in the 3' NTR of the genome of lapinized vaccine strains of CSFV. However, this study discovered that 2 T-rich insertions, 42 and 36 nt in length, are present in the viral genome of lapinized vaccine strains LPC/PRK (primary rabbit kidney) and LPC/TS (Tam-Sui), respectively. These T-rich insertions of 12, 36, and 42 nt length increases the size of PCR fragments, which are favorable genetic markers for rapid detection of and differentiation between wild-type and different lapinized vaccine strains of CSFV.

  14. Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification.

    Science.gov (United States)

    Silva, Gonçalo; Bömer, Moritz; Nkere, Chukwuemeka; Kumar, P Lava; Seal, Susan E

    2015-09-15

    Yam mosaic virus (YMV; genus Potyvirus) is considered to cause the most economically important viral disease of yams (Dioscorea spp.) in West Africa which is the dominant region for yam production globally. Yams are a vegetatively propagated crop and the use of virus-free planting material forms an essential component of disease control. Current serological and PCR-based diagnostic methods for YMV are time consuming involving a succession of target detection steps. In this study, a novel assay for specific YMV detection is described that is based on isothermal reverse transcription-recombinase polymerase amplification (RT-exoRPA). This test has been shown to be reproducible and able to detect as little as 14 pg/μl of purified RNA obtained from an YMV-infected plant, a sensitivity equivalent to that obtained with the reverse transcription-polymerase chain reaction (RT-PCR) in current general use. The RT-exoRPA assay has, however, several advantages over the RT-PCR; positive samples can be detected in less than 30 min, and amplification only requires a single incubation temperature (optimum 37°C). These features make the RT-exoRPA assay a promising candidate for adapting into a field test format to be used by yam breeding programmes or certification laboratories. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Inherited human sex reversal due to impaired nucleocytoplasmic trafficking of SRY defines a male transcriptional threshold.

    Science.gov (United States)

    Chen, Yen-Shan; Racca, Joseph D; Phillips, Nelson B; Weiss, Michael A

    2013-09-17

    Human testis determination is initiated by SRY (sex determining region on Y chromosome). Mutations in SRY cause gonadal dysgenesis with female somatic phenotype. Two subtle variants (V60L and I90M in the high-mobility group box) define inherited alleles shared by an XY sterile daughter and fertile father. Whereas specific DNA binding and bending are unaffected in a rat embryonic pre-Sertoli cell line, the variants exhibited selective defects in nucleocytoplasmic shuttling due to impaired nuclear import (V60L; mediated by Exportin-4) or export (I90M; mediated by chromosome region maintenance 1). Decreased shuttling limits nuclear accumulation of phosphorylated (activated) SRY, in turn reducing occupancy of DNA sites regulating Sertoli-cell differentiation [the testis-specific SRY-box 9 (Sox9) enhancer]. Despite distinct patterns of biochemical and cell-biological perturbations, V60L and I90M each attenuated Sox9 expression in transient transfection assays by twofold. Such attenuation was also observed in studies of V60A, a clinical variant associated with ovotestes and hence ambiguity between divergent cell fates. This shared twofold threshold is reminiscent of autosomal syndromes of transcription-factor haploinsufficiency, including XY sex reversal associated with mutations in SOX9. Our results demonstrate that nucleocytoplasmic shuttling of SRY is necessary for robust initiation of testicular development. Although also characteristic of ungulate orthologs, such shuttling is not conserved among rodents wherein impaired nuclear export of the high-mobility group box and import-dependent phosphorylation are compensated by a microsatellite-associated transcriptional activation domain. Human sex reversal due to subtle defects in the nucleocytoplasmic shuttling of SRY suggests that its transcriptional activity lies near the edge of developmental ambiguity.

  16. Comparative analysis of module-based versus direct methods for reverse-engineering transcriptional regulatory networks

    Directory of Open Access Journals (Sweden)

    Joshi Anagha

    2009-05-01

    Full Text Available Abstract Background A myriad of methods to reverse-engineer transcriptional regulatory networks have been developed in recent years. Direct methods directly reconstruct a network of pairwise regulatory interactions while module-based methods predict a set of regulators for modules of coexpressed genes treated as a single unit. To date, there has been no systematic comparison of the relative strengths and weaknesses of both types of methods. Results We have compared a recently developed module-based algorithm, LeMoNe (Learning Module Networks, to a mutual information based direct algorithm, CLR (Context Likelihood of Relatedness, using benchmark expression data and databases of known transcriptional regulatory interactions for Escherichia coli and Saccharomyces cerevisiae. A global comparison using recall versus precision curves hides the topologically distinct nature of the inferred networks and is not informative about the specific subtasks for which each method is most suited. Analysis of the degree distributions and a regulator specific comparison show that CLR is 'regulator-centric', making true predictions for a higher number of regulators, while LeMoNe is 'target-centric', recovering a higher number of known targets for fewer regulators, with limited overlap in the predicted interactions between both methods. Detailed biological examples in E. coli and S. cerevisiae are used to illustrate these differences and to prove that each method is able to infer parts of the network where the other fails. Biological validation of the inferred networks cautions against over-interpreting recall and precision values computed using incomplete reference networks. Conclusion Our results indicate that module-based and direct methods retrieve largely distinct parts of the underlying transcriptional regulatory networks. The choice of algorithm should therefore be based on the particular biological problem of interest and not on global metrics which cannot be

  17. One-step preparation of [(18)F]FPBM for PET imaging of serotonin transporter (SERT) in the brain.

    Science.gov (United States)

    Qiao, Hongwen; Zhang, Yan; Wu, Zehui; Zhu, Lin; Choi, Seok Rye; Ploessl, Karl; Kung, Hank F

    2016-08-01

    Serotonin transporters (SERT) in the brain play an important role in normal brain function. Selective serotonin reuptake inhibitors such as fluoxetine, sertraline, paroxetine, escitalopram, etc., specifically target SERT binding in the brain. Development of SERT imaging agents may be useful for studying the function of SERT by in vivo imaging. A one-step preparation of [(18)F]FPBM, 2-(2'-(dimethylamino)methyl)-4'-(3-([(18)F]fluoropropoxy)phenylthio)benzenamine, for positron emission tomography (PET) imaging of SERT binding in the brain was achieved. An active OTs intermediate, 9, was reacted with [(18)F]F(-)/K222 to produce [(18)F]FPBM in one step and in high radiochemical yield. This labeling reaction was evaluated and optimized under different temperatures, bases, solvents, and varying amounts of precursor 9. The radiolabeling reaction led to the desired [(18)F]FPBM in one step and the crude product was purified by HPLC purification to give no-carrier-added [(18)F]FPBM (radiochemical yield, 24-33%, decay corrected; radiochemical purity >99%). PET imaging studies in normal monkeys (n=4) showed fast, pronounced uptakes in the midbrain and thalamus, regions known to be rich in SERT binding sites. A displacement experiment with escitalopram (5mg/kg iv injection at 30min after [(18)F]FPBM injection) showed a rapid and complete reversal of SERT binding, suggesting that binding by [(18)F]FPBM was highly specific and reversible. A one-step radiolabeling method coupled with HPLC purification for preparation of [(18)F]FPBM was developed. Imaging studies suggest that it is feasible to use this method to prepare [(18)F]FPBM for in vivo PET imaging of SERT binding in the brain.

  18. On-demand one-step synthesis of monodisperse functional polymeric microspheres with droplet microfluidics.

    Science.gov (United States)

    Yu, Xu; Cheng, Gong; Zhou, Ming-Da; Zheng, Si-Yang

    2015-04-07

    A simple and robust method for one-step synthesis of monodisperse functional polymeric microspheres was established by generation of reversed microemulsion droplets in aqueous phase inside microfluidic chips and controlled evaporation of the organic solvent. Using this method, water-soluble nanomaterials can be easily encapsulated into biodegradable Poly(D,L-lactic-co-glycolic acid) (PLGA) to form functional microspheres. By controlling the flow rate of microemulsion phase, PLGA polymeric microspheres with narrow size distribution and diameters in the range of ∼50-100 μm were obtained. As a demonstration of the versatility of the approach, high-quality fluorescent CdTe:Zn(2+) quantum dots (QDs) of various emission spectra, superparamagnetic Fe3O4 nanoparticles, and water-soluble carbon nanotubes (CNTs) were used to synthesize fluorescent PLGA@QDs, magnetic PLGA@Fe3O4, and PLGA@CNTs polymeric microspheres, respectively. In order to show specific applications, the PLGA@Fe3O4 were modified with polydopamine (PDA), and then the silver nanoparticles grew on the surfaces of the PLGA@Fe3O4@PDA polymeric microspheres by reducting the Ag(+) to Ag(0). The as-prepared PLGA@Fe3O4@PDA-Ag microspheres showed a highly efficient catalytic reduction of the 4-nitrophenol, a highly toxic substance. The monodisperse uniform functional PLGA polymeric microspheres can potentially be critically important for multiple biomedical applications.

  19. One step gold (bio)functionalisation based on CS{sub 2}-amine reaction

    Energy Technology Data Exchange (ETDEWEB)

    Almeida, Ines [Centro de Quimica e Bioquimica, Faculdade de Ciencias da Universidade de Lisboa, Ed. C8, Campo Grande, 1749-016 Lisboa (Portugal); Cascalheira, Antonio C. [Lumisense, Lda, Campus Faculdade de Ciencias da Universidade de Lisboa, Ed. ICAT, Campo Grande, 1749-016 Lisboa (Portugal); Viana, Ana S., E-mail: anaviana@fc.ul.p [Centro de Quimica e Bioquimica, Faculdade de Ciencias da Universidade de Lisboa, Ed. C8, Campo Grande, 1749-016 Lisboa (Portugal)

    2010-12-01

    Dithiocarbamates have been regarded as alternative anchor groups to thiols on gold surfaces, and claimed to be formed in situ through the reaction between secondary amines and carbon disulphide. In this paper, we further exploit this methodology for a convenient one step biomolecule immobilisation onto gold surfaces. First, the reactivity between CS{sub 2} and electroactive compounds containing amines, primary (dopamine), secondary (epinephrine), and an amino acid (tryptophan) has been investigated by electrochemical methods. Cyclic voltammetric characterisation of the modified electrodes confirmed the immobilisation of all the target compounds, allowing the estimation of their surface concentration. The best result was obtained with epinephrine, a secondary amine, for which a typical quasi-reversible behaviour of surface confined electroactive species could be clearly depicted. Electrochemical reductive desorption studies enabled to infer on the extent of the reaction and on the relative stability of the generated monolayers. Bio-functionalisation studies have been accomplished through the reaction of CS{sub 2} with glucose oxidase in aqueous medium, and the catalytic activity of the immobilised enzyme was evaluated towards glucose, by electrochemical methods in the presence of a redox mediator. Scanning tunnelling microscopy (STM) and Atomic force microscopy (AFM) were used respectively, to characterize the gold electrodes and Glucose Oxidase coverage and distribution on the modified surfaces.

  20. One-step electrolytic preparation of Si-Fe alloys as anodes for lithium ion batteries

    Science.gov (United States)

    Wang, Hailong; Sun, Diankun; Song, Qiqi; Xie, Wenqi; Jiang, Xu; Zhang, Bo

    2016-06-01

    One-step electrolytic formation of uniform crystalline Si-Fe alloy particles was successfully demonstrated in direct electro-reduction of solid mixed oxides of SiO2 and Fe2O3 in molten CaCl2 at 900∘C. Upon constant voltage electrolysis of solid mixed oxides at 2.8V between solid oxide cathode and graphite anode for 5h, electrolytic Si-Fe with the same Si/Fe stoichimetry of the precursory oxides was generated. The firstly generated Fe could function as depolarizers to enhance reduction rate of SiO2, resulting in the enhanced reduction kinetics to the electrolysis of individual SiO2. When evaluated as anode for lithium ion batteries, the prepared SiFe electrode showed a reversible lithium storage capacity as high as 470mAh g-1 after 100 cycles at 200mA g-1, promising application in high-performance lithium ion batteries.

  1. DDX3 DEAD-Box RNA helicase inhibits hepatitis B virus reverse transcription by incorporation into nucleocapsids.

    Science.gov (United States)

    Wang, Haifeng; Kim, Seahee; Ryu, Wang-Shick

    2009-06-01

    Viruses utilize host factors in many steps of their life cycles. Yet, little is known about host factors that contribute to the life cycle of hepatitis B virus (HBV), which replicates its genome by reverse transcription. To identify host factors that contribute to viral reverse transcription, we sought to identify cellular proteins that interact with HBV polymerase (Pol) by using affinity purification coupled with mass spectrometry. One of the HBV Pol-interacting host factors identified was DDX3 DEAD-box RNA helicase, which unwinds RNA in an ATPase-dependent manner. Recently, it was shown that DDX3 is essential for both human immunodeficiency virus and hepatitis C virus infection. In contrast, we found that the ectopic expression of DDX3 led to significantly reduced viral DNA synthesis. The DDX3-mediated inhibition of viral DNA synthesis did not affect RNA encapsidation, a step prior to reverse transcription, and indicated that DDX3 inhibits HBV reverse transcription. Mutational analysis revealed that mutant DDX3 with an inactive ATPase motif, but not that with an inactive RNA helicase motif, failed to inhibit viral DNA synthesis. Our interpretation is that DDX3 inhibits viral DNA synthesis at a step following ATP hydrolysis but prior to RNA unwinding. Finally, OptiPrep density gradient analysis revealed that DDX3 was incorporated into nucleocapsids, suggesting that DDX3 inhibits viral reverse transcription following nucleocapsid assembly. Thus, DDX3 represents a novel host restriction factor that limits HBV infection.

  2. Development of a One-Step Duplex RT-PCR Method for the Simultaneous Detection of VP3/VP1 and VP1/P2B Regions of the Hepatitis A Virus.

    Science.gov (United States)

    Kim, Mi-Ju; Lee, Shin-Young; Kim, Hyun-Joong; Lee, Jeong Su; Joo, In Sun; Kwak, Hyo Sun; Kim, Hae-Yeong

    2016-08-28

    The simultaneous detection and accurate identification of hepatitis A virus (HAV) is critical in food safety and epidemiological studies to prevent the spread of HAV outbreaks. Towards this goal, a one-step duplex reverse-transcription (RT)-PCR method was developed targeting the VP1/P2B and VP3/VP1 regions of the HAV genome for the qualitative detection of HAV. An HAV RT-qPCR standard curve was produced for the quantification of HAV RNA. The detection limit of the duplex RT-PCR method was 2.8 × 10(1) copies of HAV. The PCR products enabled HAV genotyping analysis through DNA sequencing, which can be applied for epidemiological investigations. The ability of this duplex RT-PCR method to detect HAV was evaluated with HAV-spiked samples of fresh lettuce, frozen strawberries, and oysters. The limit of detection of the one-step duplex RT-PCR for each food model was 9.4 × 10(2) copies/20 g fresh lettuce, 9.7 × 10(3) copies/20 g frozen strawberries, and 4.1 × 10(3) copies/1.5 g oysters. Use of a one-step duplex RT-PCR method has advantages such as shorter time, decreased cost, and decreased labor owing to the single amplification reaction instead of four amplifications necessary for nested RT-PCR.

  3. Fate of HIV-1 cDNA intermediates during reverse transcription is dictated by transcription initiation site of virus genomic RNA

    Science.gov (United States)

    Masuda, Takao; Sato, Yoko; Huang, Yu-Lun; Koi, Satoshi; Takahata, Tatsuro; Hasegawa, Atsuhiko; Kawai, Gota; Kannagi, Mari

    2015-01-01

    Retroviral reverse transcription is accomplished by sequential strand-transfers of partial cDNA intermediates copied from viral genomic RNA. Here, we revealed an unprecedented role of 5′-end guanosine (G) of HIV-1 genomic RNA for reverse transcription. Based on current consensus for HIV-1 transcription initiation site, HIV-1 transcripts possess a single G at 5′-ends (G1-form). However, we found that HIV-1 transcripts with additional Gs at 5′-ends (G2- and G3-forms) were abundantly expressed in infected cells by using alternative transcription initiation sites. The G2- and G3-forms were also detected in the virus particle, although the G1-form predominated. To address biological impact of the 5′-G number, we generated HIV clone DNA to express the G1-form exclusively by deleting the alternative initiation sites. Virus produced from the clone showed significantly higher strand-transfer of minus strong-stop cDNA (-sscDNA). The in vitro assay using synthetic HIV-1 RNAs revealed that the abortive forms of -sscDNA were abundantly generated from the G3-form RNA, but dramatically reduced from the G1-form. Moreover, the strand-transfer of -sscDNA from the G1-form was prominently stimulated by HIV-1 nucleocapsid. Taken together, our results demonstrated that the 5′-G number that corresponds to HIV-1 transcription initiation site was critical for successful strand-transfer of -sscDNA during reverse transcription. PMID:26631448

  4. Intracytoplasmic maturation of the human immunodeficiency virus type 1 reverse transcription complexes determines their capacity to integrate into chromatin

    Directory of Open Access Journals (Sweden)

    Kashanchi Fatah

    2006-01-01

    Full Text Available Abstract Background The early events of the HIV-1 life cycle include entry of the viral core into target cell, assembly of the reverse transcription complex (RTCs performing reverse transcription, its transformation into integration-competent complexes called pre-integration complexes (PICs, trafficking of complexes into the nucleus, and finally integration of the viral DNA into chromatin. Molecular details and temporal organization of these processes remain among the least investigated and most controversial problems in the biology of HIV. Results To quantitatively evaluate maturation and nuclear translocation of the HIV-1 RTCs, nucleoprotein complexes isolated from the nucleus (nRTC and cytoplasm (cRTC of HeLa cells infected with MLV Env-pseudotyped HIV-1 were analyzed by real-time PCR. While most complexes completed reverse transcription in the cytoplasm, some got into the nucleus before completing DNA synthesis. The HIV-specific RNA complexes could get into the nucleus when reverse transcription was blocked by reverse transcriptase inhibitor, although nuclear import of RNA complexes was less efficient than of DNA-containing RTCs. Analysis of the RTC nuclear import in synchronized cells infected in the G2/M phase of the cell cycle showed enrichment in the nuclei of RTCs containing incomplete HIV-1 DNA compared to non-synchronized cells, where RTCs with complete reverse transcripts prevailed. Immunoprecipitation assays identified viral proteins IN, Vpr, MA, and cellular Ini1 and PML associated with both cRTCs and nRTCs, whereas CA was detected only in cRTCs and RT was diminished in nRTCs. Cytoplasmic maturation of the complexes was associated with increased immunoreactivity with anti-Vpr and anti-IN antibodies, and decreased reactivity with antibodies to RT. Both cRTCs and nRTCs carried out endogenous reverse transcription reaction in vitro. In contrast to cRTCs, in vitro completion of reverse transcription in nRTCs did not increase their

  5. Effect of evaporation of solvents from one-step, self-etching adhesives

    DEFF Research Database (Denmark)

    Furuse, Adilson Yoshio; Peutzfeldt, Anne; Asmussen, Erik

    2008-01-01

    PURPOSE: To investigate whether and to what extent the bonding capacity of one-step, self-etching adhesives is influenced by the degree to which solvent is evaporated. MATERIALS AND METHODS: Seven one-step, self-etching adhesives were tested (Adper Prompt L-Pop, Clearfil S3 Bond, Futurabond NR, G...

  6. Typing of Poultry Influenza Virus (H5 and H7 by Reverse Transcription- Polymerase Chain Reaction

    Directory of Open Access Journals (Sweden)

    Cesare Bonacina

    2010-01-01

    Full Text Available The ability of the influenza Orthomixovirus to undergo to continually antigenically changes that can affect its pathogenicity and its diffusion, explains the growing seriousness of this disease and the recent epizoozies in various parts of the world. There have been 15 HA and 9 NA type A sub-types of the influenza virus identified all of which are present in birds. Until now the very virulent avian influenza viruses identified were all included to the H5 and H7 sub-types. We here show that is possible to identify the H5 and H7 sub-types with reverse transcription-polymerase chain reaction (RT-PCR by using a set of specific primers for each HA sub-type. The RT-PCR is a quick and sensitive method of identifying the HA sub-types of the influenza virus directly from homogenised organs.

  7. Application of anti-listerial bacteriocins: monitoring enterocin expression by multiplex relative reverse transcription-PCR.

    Science.gov (United States)

    Williams, D Ross; Chanos, Panagiotis

    2012-12-01

    Listeriosis is a deadly food-borne disease, and its incidence may be limited through the biotechnological exploitation of a number of anti-listerial biocontrol agents. The most widely used of these agents are bacteriocins and the Class II enterocins are characterized by their activity against Listeria. Enterocins are primarily produced by enterococci, particularly Enterococcus faecium and many strains have been described, often encoding multiple bacteriocins. The use of these strains in food will require that they are free of virulence functions and that they exhibit a high level expression of anti-listerial enterocins in fermentation conditions. Multiplex relative RT (reverse transcription)-PCR is a technique that is useful in the discovery of advantageous expression characteristics among enterocin-producing strains. It allows the levels of individual enterocin gene expression to be monitored and determination of how expression is altered under different growth conditions.

  8. Distinct transcriptional networks in quiescent myoblasts: a role for Wnt signaling in reversible vs. irreversible arrest.

    Science.gov (United States)

    Subramaniam, Sindhu; Sreenivas, Prethish; Cheedipudi, Sirisha; Reddy, Vatrapu Rami; Shashidhara, Lingadahalli Subrahmanya; Chilukoti, Ravi Kumar; Mylavarapu, Madhavi; Dhawan, Jyotsna

    2014-01-01

    Most cells in adult mammals are non-dividing: differentiated cells exit the cell cycle permanently, but stem cells exist in a state of reversible arrest called quiescence. In damaged skeletal muscle, quiescent satellite stem cells re-enter the cell cycle, proliferate and subsequently execute divergent programs to regenerate both post-mitotic myofibers and quiescent stem cells. The molecular basis for these alternative programs of arrest is poorly understood. In this study, we used an established myogenic culture model (C2C12 myoblasts) to generate cells in alternative states of arrest and investigate their global transcriptional profiles. Using cDNA microarrays, we compared G0 myoblasts with post-mitotic myotubes. Our findings define the transcriptional program of quiescent myoblasts in culture and establish that distinct gene expression profiles, especially of tumour suppressor genes and inhibitors of differentiation characterize reversible arrest, distinguishing this state from irreversibly arrested myotubes. We also reveal the existence of a tissue-specific quiescence program by comparing G0 C2C12 myoblasts to isogenic G0 fibroblasts (10T1/2). Intriguingly, in myoblasts but not fibroblasts, quiescence is associated with a signature of Wnt pathway genes. We provide evidence that different levels of signaling via the canonical Wnt pathway characterize distinct cellular states (proliferation vs. quiescence vs. differentiation). Moderate induction of Wnt signaling in quiescence is associated with critical properties such as clonogenic self-renewal. Exogenous Wnt treatment subverts the quiescence program and negatively affects clonogenicity. Finally, we identify two new quiescence-induced regulators of canonical Wnt signaling, Rgs2 and Dkk3, whose induction in G0 is required for clonogenic self-renewal. These results support the concept that active signal-mediated regulation of quiescence contributes to stem cell properties, and have implications for pathological

  9. On-chip single-copy real-time reverse-transcription PCR in isolated picoliter droplets

    Energy Technology Data Exchange (ETDEWEB)

    Beer, N R; Wheeler, E; Lee-Houghton, L; Watkins, N; Nasarabadi, S; Hebert, N; Leung, P; Arnold, D; Bailey, C; Colston, B

    2007-12-19

    The first lab-on-chip system for picoliter droplet generation and RNA isolation, followed by reverse transcription, and PCR amplification with real-time fluorescence detection in the trapped droplets has been developed. The system utilized a shearing T-junction in a fused silica device to generate a stream of monodisperse picoliter-scale droplets that were isolated from the microfluidic channel walls and each other by the oil phase carrier. An off-chip valving system stopped the droplets on-chip, allowing thermal cycling for reverse transcription and subsequent PCR amplification without droplet motion. This combination of the established real-time reverse transcription-PCR assay with digital microfluidics is ideal for isolating single-copy RNA and virions from a complex environment, and will be useful in viral discovery and gene-profiling applications.

  10. Detection of Cardamom mosaic virus and Banana bract mosaic virus in cardamom using SYBR Green based reverse transcription-quantitative PCR.

    Science.gov (United States)

    Siljo, A; Bhat, A I; Biju, C N

    2014-01-01

    Cardamom being perennial, propagated vegetatively, detecting viruses in planting material is important to check the spread of viruses through infected material. Thus development of effective and sensitive assay for detection of viruses is need of the time. In this view, assay for the detection of Cardamom mosaic virus (CdMV) and Banana bract mosaic virus (BBrMV), infecting cardamom was developed using SYBR Green one step reverse transcription-quantitative PCR (RT-qPCR). The RT-qPCR assay amplified all isolates of CdMV and BBrMV tested but no amplification was obtained with RNA of healthy plants. Recombinant plasmids carrying target virus regions corresponding to both viruses were quantified, serially diluted and used as standards in qPCR to develop standard curve to enable quantification. When tenfold serial dilutions of the total RNAs from infected plants were tested through RT-qPCR, the detection limit of the assay was estimated to be 16 copies for CdMV and 10 copies for BBrMV, which was approximately 1,000-fold higher than the conventional RT-PCR. The RT-qPCR assay was validated by testing field samples collected from different cardamom growing regions of India. This is the first report of RT-qPCR assay for the detection of CdMV and BBrMV in cardamom.

  11. Standardized positive controls for detection of norovirus by reverse transcription PCR

    Directory of Open Access Journals (Sweden)

    Oh SeHwan

    2011-05-01

    Full Text Available Abstract Background Norovirus is one of the most common causes of nonbacterial gastroenteritis in humans. Rapid spread by contaminated food and person-to-person transmission through the fecal-oral route are characteristics of norovirus epidemiology and result in high morbidity in vulnerable patient populations. Therefore, detection of norovirus is a major public health concern. Currently, the most common method for detecting and differentiating among norovirus strains in clinical and environmental samples is reverse transcription PCR (RT-PCR. Standardized positive controls used in RT-PCR assays to detect norovirus are designed to overcome the problem of false-negative results due to PCR inhibitors and suboptimal reaction conditions. Results In the current study, four types of RNA transcripts were produced from plasmids: norovirus GI-5 and GII-4 capsid regions with human rotavirus (VP7 gene derived fragment insertions, and norovirus GI-6 and GII-4 capsid regions with hepatitis A virus (VP1/P2A gene derived fragment insertions. These size-distinguishable products were used as positive controls under the RT-PCR assay conditions used to detect NoV in stool and groundwater samples. Their reliability and reproducibility was confirmed by multiple sets of experiments. Conclusions These standardized products may contribute to the reliable and accurate diagnosis by RT-PCR of norovirus outbreaks, when conducted by laboratories located in different regions.

  12. The reverse transcription inhibitor abacavir shows anticancer activity in prostate cancer cell lines.

    Directory of Open Access Journals (Sweden)

    Francesca Carlini

    Full Text Available BACKGROUND: Transposable Elements (TEs comprise nearly 45% of the entire genome and are part of sophisticated regulatory network systems that control developmental processes in normal and pathological conditions. The retroviral/retrotransposon gene machinery consists mainly of Long Interspersed Nuclear Elements (LINEs-1 and Human Endogenous Retroviruses (HERVs that code for their own endogenous reverse transcriptase (RT. Interestingly, RT is typically expressed at high levels in cancer cells. Recent studies report that RT inhibition by non-nucleoside reverse transcriptase inhibitors (NNRTIs induces growth arrest and cell differentiation in vitro and antagonizes growth of human tumors in animal model. In the present study we analyze the anticancer activity of Abacavir (ABC, a nucleoside reverse transcription inhibitor (NRTI, on PC3 and LNCaP prostate cancer cell lines. PRINCIPAL FINDINGS: ABC significantly reduces cell growth, migration and invasion processes, considerably slows S phase progression, induces senescence and cell death in prostate cancer cells. Consistent with these observations, microarray analysis on PC3 cells shows that ABC induces specific and dose-dependent changes in gene expression, involving multiple cellular pathways. Notably, by quantitative Real-Time PCR we found that LINE-1 ORF1 and ORF2 mRNA levels were significantly up-regulated by ABC treatment. CONCLUSIONS: Our results demonstrate the potential of ABC as anticancer agent able to induce antiproliferative activity and trigger senescence in prostate cancer cells. Noteworthy, we show that ABC elicits up-regulation of LINE-1 expression, suggesting the involvement of these elements in the observed cellular modifications.

  13. One-step discrimination scheme on N-particle Greenberger-Horne-Zeilinger bases

    Institute of Scientific and Technical Information of China (English)

    Wang Xin-Wen; Liu Xiang; Fang Mao-Fa

    2007-01-01

    We present an experimentally feasible one-step discrimination scheme on Bell bases with trapped ions, and then generalize it to the case of N-ion Greenberger-Horne-Zeilinger (GHZ) bases. In the scheme, all the orthogonal and complete N-ion GHZ internal states can be exactly discriminated only by one step, and thus it takes very short time. Moreover, the scheme is insensitive to thermal motion and dose not require the individual addressing of the ions. The Bell-state and GHZ-state one-step discrimination scheme can be widely used in quantum information processing based on ion-trap set-up.

  14. Detection of EWS-FLI1 fusion transcripts in paraffin embedded tissues of peripheral primitive neuroectodermal tumors by nested reverse transcription polymerase chain reaction

    Institute of Scientific and Technical Information of China (English)

    Qixing Gong; Qinhe Fan; Zhihong Zhang; Weiming Zhang

    2005-01-01

    Objective: To assess the feasibility and significance of detecting EWS-FLIlfusion transcripts in paraffin embedded tissues of peripheral primitive neuroectodermal tumors (PNETs) by nested reverse transcription polymerase chain reaction (RT-PCR).Methods: Twelve formalin-fixed and paraffin-embedded (FFPE) samples of PNET were retrieved from archive and consultation materials,together with eight cases of controlled tumor. EWS-FLI1 fusion transcripts were detected by nested RT-PCR. Home-keeping gene β-actin was used to detect the quality of mRNA. Results: β-actin mRNA was detected in 9 of the 12 tumor cases. EWS-FLI1 fusion transcripts were detected in 6 cases, among which 4 had a "type 1" fusion transcript and 2 had a "type 2" fusion transcript. None of the controlled tumor was detected the fusion gene. Conclusion: RT-PCR is a feasible method for the detection of EWS-FLI1 fusion transcripts in FFPE tissues in PNET and the result is meaningful in differential diagnosis and prognostic evaluation.

  15. Are one-step adhesives easier to use and better performing? Multifactorial assessment of contemporary one-step self-etching adhesives.

    Science.gov (United States)

    Van Landuyt, Kirsten L; Mine, Atsushi; De Munck, Jan; Jaecques, Siegfried; Peumans, Marleen; Lambrechts, Paul; Van Meerbeek, Bart

    2009-06-01

    The objective of this study was to examine whether one-step self-etching adhesives (1-SEAs) really have an advantage over multistep systems. Nine one-step self-etching adhesives (Absolute, Adper Prompt L-Pop, Clearfil S3 Bond, G-Bond, Hybrid Bond, iBond, One-up Bond F Plus, Optibond All-in-one and Xeno III) were included in this study. One two-step self-etching adhesive (Clearfil SE Bond) and one three-step etch-and-rinse adhesive (Optibond FL) served as controls. Their microtensile bond strength to bur-cut enamel and dentin was determined using a standardized protocol and the respective adhesive/dentin interface of these adhesives was characterized by transmission electron microscopy. Statistical analysis was performed with the Kruskal-Wallis nonparametric test. Regarding bond strength, the control adhesives tended to perform superior to the one-step adhesives. However, a significant difference between the control adhesives and some one-step adhesives could not always be demonstrated, partly due to the statistical setup of this study. Interface analysis by electron microscopy showed wide variation among the one-step adhesives, depending on their composition and their acidity. 1-SEAs also exhibited two different kinds of droplets, depending on their hydrophilicity. Hydrophobic HEMA-free 1-SEAs such as G-Bond were prone to phase separation, while especially HEMA-containing hydrophilic 1-SEAs, such as Clearfil S3 Bond and Xeno III were predisposed to forming osmosis-induced droplets. Hybrid bond, Absolute, and iBond featured both phase separation as well as osmosis. Optibond All-in-one exhibited a clustering reaction of the filler particles upon solvent evaporation. All adhesives including the control adhesives showed signs of nanoleakage, indicating that all adhesives are to some extent permeable to water. A definitive conclusion with regard to quantitative assessment of nanoleakage was much hindered by inconsistencies in the silver deposition. The application

  16. Rapid and Specific Detection of tdh, trh1, and trh2 mRNA of Vibrio parahaemolyticus by Transcription-Reverse Transcription Concerted Reaction with an Automated System

    OpenAIRE

    Nakaguchi, Yoshitsugu; Ishizuka, Tetsuya; Ohnaka, Satoru; Hayashi, Toshinori; Yasukawa, Kiyoshi; Ishiguro, Takahiko; Nishibuchi, Mitsuaki

    2004-01-01

    Vibrio parahaemolyticus strains carrying the thermostable direct hemolysin (TDH) tdh gene, the TDH-related hemolysin (trh) gene, or both genes are considered virulent strains. We previously demonstrated that the transcription-reverse transcription concerted (TRC) method could be used to quantify the amount of mRNA transcribed from the tdh gene by using an automated detection system. In this study, we devised two TRC-based assays to quantify the mRNAs transcribed from the trh1 and trh2 genes, ...

  17. A Modified Reverse One-Hybrid Screen Identifies Transcriptional Activation Domains in PHYTOCHROME-INTERACTING FACTOR 3.

    Science.gov (United States)

    Dalton, Jutta C; Bätz, Ulrike; Liu, Jason; Curie, Gemma L; Quail, Peter H

    2016-01-01

    Transcriptional activation domains (TADs) are difficult to predict and identify, since they are not conserved and have little consensus. Here, we describe a yeast-based screening method that is able to identify individual amino acid residues involved in transcriptional activation in a high throughput manner. A plant transcriptional activator, PIF3 (phytochrome interacting factor 3), was fused to the yeast GAL4-DNA-binding Domain (BD), driving expression of the URA3 (Orotidine 5'-phosphate decarboxylase) reporter, and used for negative selection on 5-fluroorotic acid (5FOA). Randomly mutagenized variants of PIF3 were then selected for a loss or reduction in transcriptional activation activity by survival on FOA. In the process, we developed a strategy to eliminate false positives from negative selection that can be used for both reverse-1- and 2-hybrid screens. With this method we were able to identify two distinct regions in PIF3 with transcriptional activation activity, both of which are functionally conserved in PIF1, PIF4, and PIF5. Both are collectively necessary for full PIF3 transcriptional activity, but neither is sufficient to induce transcription autonomously. We also found that the TAD appear to overlap physically with other PIF3 functions, such as phyB binding activity and consequent phosphorylation. Our protocol should provide a valuable tool for identifying, analyzing and characterizing novel TADs in eukaryotic transcription factors, and thus potentially contribute to the unraveling of the mechanism underlying transcriptional activation.

  18. A Modified Reverse One-Hybrid Screen Identifies Transcriptional Activation Domains in PHYTOCHROME-INTERACTING FACTOR 3

    Science.gov (United States)

    Dalton, Jutta C.; Bätz, Ulrike; Liu, Jason; Curie, Gemma L.; Quail, Peter H.

    2016-01-01

    Transcriptional activation domains (TADs) are difficult to predict and identify, since they are not conserved and have little consensus. Here, we describe a yeast-based screening method that is able to identify individual amino acid residues involved in transcriptional activation in a high throughput manner. A plant transcriptional activator, PIF3 (phytochrome interacting factor 3), was fused to the yeast GAL4-DNA-binding Domain (BD), driving expression of the URA3 (Orotidine 5′-phosphate decarboxylase) reporter, and used for negative selection on 5-fluroorotic acid (5FOA). Randomly mutagenized variants of PIF3 were then selected for a loss or reduction in transcriptional activation activity by survival on FOA. In the process, we developed a strategy to eliminate false positives from negative selection that can be used for both reverse-1- and 2-hybrid screens. With this method we were able to identify two distinct regions in PIF3 with transcriptional activation activity, both of which are functionally conserved in PIF1, PIF4, and PIF5. Both are collectively necessary for full PIF3 transcriptional activity, but neither is sufficient to induce transcription autonomously. We also found that the TAD appear to overlap physically with other PIF3 functions, such as phyB binding activity and consequent phosphorylation. Our protocol should provide a valuable tool for identifying, analyzing and characterizing novel TADs in eukaryotic transcription factors, and thus potentially contribute to the unraveling of the mechanism underlying transcriptional activation. PMID:27379152

  19. Detection of Zika Virus in Desiccated Mosquitoes by Real-Time Reverse Transcription PCR and Plaque Assay

    Science.gov (United States)

    Savage, Harry M.

    2017-01-01

    We assayed Zika virus–infected mosquitoes stored at room temperature for <30 days for live virus by using plaque assay and virus RNA by using real-time reverse transcription PCR. Viable virus was detected in samples stored <10 days, and virus RNA was detected in samples held for 30 days. PMID:28075325

  20. Rapid and specific detection of Lassa virus by reverse transcription-PCR coupled with oligonucleotide array hybridization.

    Science.gov (United States)

    Olschläger, Stephan; Günther, Stephan

    2012-07-01

    To facilitate sequence-specific detection of DNA amplified in a diagnostic reverse transcription (RT)-PCR for Lassa virus, we developed an array featuring 47 oligonucleotide probes for post-PCR hybridization of the amplicons. The array procedure may be performed with low-tech equipment and does not take longer than agarose gel detection.

  1. One-step apexification using platelet rich fibrin matrix and mineral trioxide aggregate apical barrier

    Directory of Open Access Journals (Sweden)

    Anisha Kumar

    2014-01-01

    In one-step apexification using MTA, the technical problem encountered is controlling the overfill or underfill of MTA. The use of a matrix material helps to overcome this shortcoming. Platelet rich fibrin (PRF is an immune platelet concentrate, which can be used as a matrix, it also promotes wound healing and repair. This case report presents a case of one step apexification using MTA as an apical barrier and autologous PRF as an internal matrix.

  2. A Simple Reverse Transcription-Polymerase Chain Reaction for Dengue Type 2 Virus Identification

    Directory of Open Access Journals (Sweden)

    Figueiredo Luiz Tadeu M

    1997-01-01

    Full Text Available We show here a simplified reverse transcription-polymerase chain reaction (RT-PCR for identification of dengue type 2 virus. Three dengue type 2 virus strains, isolated from Brazilian patients, and yellow fever vaccine 17DD, as a negative control, were used in this study. C6/36 cells were infected with the virus, and tissue culture fluids were collected after 7 days of infection period. The RT-PCR, a combination of RT and PCR done after a single addition of reagents in a single reaction vessel was carried out following a digestion of virus with 1% Nonidet P-40. The 50ml assay reaction mixture included 50 pmol of a dengue type 2 specific primer pair amplifying a 210 base pair sequence of the envelope protein gene, 0.1 mM of the four deoxynucleoside triphosphates, 7.5U of reverse transcriptase, and 1U of thermostable Taq DNA polymerase. The reagent mixture was incubated for 15 min at 37oC for RT followed by a variable amount of cycles of two-step PCR amplification (92oC for 60 sec, 53oC for 60 sec with slow temperature increment. The PCR products were subjected to 1.7% agarose gel electrophoresis and visualized with UV light after gel incubation in ethidium bromide solution. DNA bands were observed after 25 and 30 cycles of PCR. Virus amount as low as 102.8 TCID50/ml was detected by RT-PCR. Specific DNA amplification was observed with the three dengue type 2 strains. This assay has advantages compared to other RT-PCRs: it avoids laborious extraction of virus RNA; the combination of RT and PCR reduces assay time, facilitates the performance and reduces risk of contamination; the two-step PCR cycle produces a clear DNA amplification, saves assay time and simplifies the technique

  3. Early and transient reverse transcription during primary deltaretroviral infection of sheep

    Directory of Open Access Journals (Sweden)

    Wattel Eric

    2008-02-01

    Full Text Available Abstract Background Intraindividual genetic variability plays a central role in deltaretrovirus replication and associated leukemogenesis in animals as in humans. To date, the replication of these viruses has only been investigated during the chronic phase of the infection when they mainly spread through the clonal expansion of their host cells, vary through a somatic mutation process without evidence for reverse transcriptase (RT-associated substitution. Primary infection of a new organism necessary involves allogenic cell infection and thus reverse transcription. Results Here we demonstrate that the primary experimental bovine leukemia virus (BLV infection of sheep displays an early and intense burst of horizontal replicative dissemination of the virus generating frequent RT-associated substitutions that account for 69% of the in vivo BLV genetic variability during the first 8 months of the infection. During this period, evidence has been found of a cell-to-cell passage of a mutated sequence and of a sequence having undergone both RT-associated and somatic mutations. The detection of RT-dependent proviral substitution was restricted to a narrow window encompassing the first 250 days following seroconversion. Conclusion In contrast to lentiviruses, deltaretroviruses display two time-dependent mechanisms of genetic variation that parallel their two-step nature of replication in vivo. We propose that the early and transient RT-based horizontal replication helps the virus escape the first wave of host immune response whereas somatic-dependent genetic variability during persistent clonal expansion helps infected clones escape the persistent and intense immune pressure that characterizes the chronic phase of deltaretrovirus infection.

  4. Sensitive Detection of Measles Virus Infection in the Blood and Tissues of Humanized Mouse by One-step Quantitative RT-PCR

    Directory of Open Access Journals (Sweden)

    Shota eIkeno

    2013-10-01

    Full Text Available Live attenuated measles virus (MV has long been recognized as a safe and effective vaccine, and it has served as the basis for development of various MV-based vaccines. However, because MV is a human-tropic virus, the evaluation of MV-based vaccines has been hampered by the lack of a small-animal model. The humanized mouse, a recently developed system in which an immunodeficient mouse is transplanted with human fetal tissues or hematopoietic stem cells, may represent a suitable model. Here, we developed a sensitive one-step quantitative reverse transcription (qRT PCR that simultaneously measures nucleocapsid (N and human RNase P mRNA levels. The results can be used to monitor MV infection in a humanized mouse model. Using this method, we elucidated the replication kinetics of MV expressing EGFP both in vitro and in humanized mice in parallel with flow-cytometric analysis. Because our qRT-PCR system was sensitive enough to detect MV expression using RNA extracted from a small number of cells, it can be used to monitor MV infection in humanized mice by sequential blood sampling.

  5. Diagnosis of Cetacean morbillivirus: A sensitive one step real time RT fast-PCR method based on SYBR(®) Green.

    Science.gov (United States)

    Sacristán, Carlos; Carballo, Matilde; Muñoz, María Jesús; Bellière, Edwige Nina; Neves, Elena; Nogal, Verónica; Esperón, Fernando

    2015-12-15

    Cetacean morbillivirus (CeMV) (family Paramyxoviridae, genus Morbillivirus) is considered the most pathogenic virus of cetaceans. It was first implicated in the bottlenose dolphin (Tursiops truncatus) mass stranding episode along the Northwestern Atlantic coast in the late 1980s, and in several more recent worldwide epizootics in different Odontoceti species. This study describes a new one step real-time reverse transcription fast polymerase chain reaction (real-time RT-fast PCR) method based on SYBR(®) Green to detect a fragment of the CeMV fusion protein gene. This primer set also works for conventional RT-PCR diagnosis. This method detected and identified all three well-characterized strains of CeMV: porpoise morbillivirus (PMV), dolphin morbillivirus (DMV) and pilot whale morbillivirus (PWMV). Relative sensitivity was measured by comparing the results obtained from 10-fold dilution series of PMV and DMV positive controls and a PWMV field sample, to those obtained by the previously described conventional phosphoprotein gene based RT-PCR method. Both the conventional and real-time RT-PCR methods involving the fusion protein gene were 100- to 1000-fold more sensitive than the previously described conventional RT-PCR method.

  6. Specific detection of H5N1 avian influenza A virus in field specimens by a one-step RT-PCR assay

    Directory of Open Access Journals (Sweden)

    Gupta Sanjay

    2006-03-01

    Full Text Available Abstract Background Continuous outbreaks of the highly pathogenic H5N1 avian influenza A in Asia has resulted in an urgent effort to improve current diagnostics to aid containment of the virus and lower the threat of a influenza pandemic. We report here the development of a PCR-based assay that is highly specific for the H5N1 avian influenza A virus. Methods A one-step reverse-transcription PCR assay was developed to detect the H5N1 avian influenza A virus. The specificity of the assay was shown by testing sub-types of influenza A virus and other viral and bacterial pathogens; and on field samples. Results Validation on 145 field specimens from Vietnam and Malaysia showed that the assay was specific without cross reactivity to a number of other infuenza strains as well as human respiratory related pathogens. Detection was 100% from allantoic fluid in H5N1 positive samples, suggesting it to be a reliable sampling source for accurate detection. Conclusion The assay developed from this study indicates that the primers are specific for the H5N1 influenza virus. As shown by the field tested results, this assay would be highly useful as a diagnostic tool to help identify and control influenza epidemics.

  7. Multiplex, Quantitative, Reverse Transcription PCR Detection of Influenza Viruses Using Droplet Microfluidic Technology

    Directory of Open Access Journals (Sweden)

    Ravi Prakash

    2014-12-01

    Full Text Available Quantitative, reverse transcription, polymerase chain reaction (qRT-PCR is facilitated by leveraging droplet microfluidic (DMF system, which due to its precision dispensing and sample handling capabilities at microliter and lower volumes has emerged as a popular method for miniaturization of the PCR platform. This work substantially improves and extends the functional capabilities of our previously demonstrated single qRT-PCR micro-chip, which utilized a combination of electrostatic and electrowetting droplet actuation. In the reported work we illustrate a spatially multiplexed micro-device that is capable of conducting up to eight parallel, real-time PCR reactions per usage, with adjustable control on the PCR thermal cycling parameters (both process time and temperature set-points. This micro-device has been utilized to detect and quantify the presence of two clinically relevant respiratory viruses, Influenza A and Influenza B, in human samples (nasopharyngeal swabs, throat swabs. The device performed accurate detection and quantification of the two respiratory viruses, over several orders of RNA copy counts, in unknown (blind panels of extracted patient samples with acceptably high PCR efficiency (>94%. The multi-stage qRT-PCR assays on eight panel patient samples were accomplished within 35–40 min, with a detection limit for the target Influenza virus RNAs estimated to be less than 10 RNA copies per reaction.

  8. Reverse transcriptase genes are highly abundant and transcriptionally active in marine plankton assemblages

    KAUST Repository

    Lescot, Magali

    2015-11-27

    Genes encoding reverse transcriptases (RTs) are found in most eukaryotes, often as a component of retrotransposons, as well as in retroviruses and in prokaryotic retroelements. We investigated the abundance, classification and transcriptional status of RTs based on Tara Oceans marine metagenomes and metatranscriptomes encompassing a wide organism size range. Our analyses revealed that RTs predominate large-size fraction metagenomes (>5 μm), where they reached a maximum of 13.5% of the total gene abundance. Metagenomic RTs were widely distributed across the phylogeny of known RTs, but many belonged to previously uncharacterized clades. Metatranscriptomic RTs showed distinct abundance patterns across samples compared with metagenomic RTs. The relative abundances of viral and bacterial RTs among identified RT sequences were higher in metatranscriptomes than in metagenomes and these sequences were detected in all metatranscriptome size fractions. Overall, these observations suggest an active proliferation of various RT-assisted elements, which could be involved in genome evolution or adaptive processes of plankton assemblage.

  9. Reverse transcriptase genes are highly abundant and transcriptionally active in marine plankton assemblages.

    Science.gov (United States)

    Lescot, Magali; Hingamp, Pascal; Kojima, Kenji K; Villar, Emilie; Romac, Sarah; Veluchamy, Alaguraj; Boccara, Martine; Jaillon, Olivier; Iudicone, Daniele; Bowler, Chris; Wincker, Patrick; Claverie, Jean-Michel; Ogata, Hiroyuki

    2016-05-01

    Genes encoding reverse transcriptases (RTs) are found in most eukaryotes, often as a component of retrotransposons, as well as in retroviruses and in prokaryotic retroelements. We investigated the abundance, classification and transcriptional status of RTs based on Tara Oceans marine metagenomes and metatranscriptomes encompassing a wide organism size range. Our analyses revealed that RTs predominate large-size fraction metagenomes (>5 μm), where they reached a maximum of 13.5% of the total gene abundance. Metagenomic RTs were widely distributed across the phylogeny of known RTs, but many belonged to previously uncharacterized clades. Metatranscriptomic RTs showed distinct abundance patterns across samples compared with metagenomic RTs. The relative abundances of viral and bacterial RTs among identified RT sequences were higher in metatranscriptomes than in metagenomes and these sequences were detected in all metatranscriptome size fractions. Overall, these observations suggest an active proliferation of various RT-assisted elements, which could be involved in genome evolution or adaptive processes of plankton assemblage.

  10. Normalization of Reverse Transcription Quantitative PCR Data During Ageing in Distinct Cerebral Structures.

    Science.gov (United States)

    Bruckert, G; Vivien, D; Docagne, F; Roussel, B D

    2016-04-01

    Reverse transcription quantitative-polymerase chain reaction (RT-qPCR) has become a routine method in many laboratories. Normalization of data from experimental conditions is critical for data processing and is usually achieved by the use of a single reference gene. Nevertheless, as pointed by the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines, several reference genes should be used for reliable normalization. Ageing is a physiological process that results in a decline of many expressed genes. Reliable normalization of RT-qPCR data becomes crucial when studying ageing. Here, we propose a RT-qPCR study from four mouse brain regions (cortex, hippocampus, striatum and cerebellum) at different ages (from 8 weeks to 22 months) in which we studied the expression of nine commonly used reference genes. With the use of two different algorithms, we found that all brain structures need at least two genes for a good normalization step. We propose specific pairs of gene for efficient data normalization in the four brain regions studied. These results underline the importance of reliable reference genes for specific brain regions in ageing.

  11. Diagnosis of Brazilian vesiculoviruses by reverse transcription-polymerase chain reaction

    Directory of Open Access Journals (Sweden)

    Daniela Wey Bonutti

    2005-04-01

    Full Text Available We describe a reverse transcription-polymerase chain reaction (RT-PCR and a nested-PCR for diagnosis of Piry, Carajás, Cocal, and Alagoas vesiculoviruses from Brazil. The RNA extracts of viral and clinical samples were submitted to a RT-PCR using Vesiculovirus G primers that amplify part of the glycoprotein gene. The RT-PCR produced amplicons of expected size, 290 base pair, for the four studied viruses. The RT-PCR showed a high sensitivity being 151.3 times (2.18 log more sensitive for the detection of Piry virus than the classical procedure for virus detection in tissue culture based on the viral cytophatic effect. Amplicons had nucleotides sequenced and were aligned in order to select internal primers for a nested-PCR to confirm the origin of Piry, Carajás, Cocal, and Alagoas Vesiculovirus. Ten blood and tarsal pad epithelial samples of infected Guinea-pigs had Vesiculovirus genome amplified by RT-nested-PCR.

  12. Detection of HCV-RNA by Reverse Transcription Polymerase Chain Reaction Using Biotinylated and Radioiodinated Primers

    Energy Technology Data Exchange (ETDEWEB)

    Ryu, Jin Sook; Moon, Dae Hyuk; Cheon, Jun Hong; Chung, Yoon Young; Park, Hung Dong; Chung, Young Hwa; Lee, Young Sang [Asan Medical Center, University of Ulsan, Seoul (Korea, Republic of)

    1994-07-15

    This study was performed to evaluate the clinical applicability of the reverse transcription polymerase chain reaction (RT-PCR) kit of HCV-RNA using biotinylated and radioiodinated primers. Study subjects were 118 patients with positive anti-HCV. HCV-RNA in patients serum was extracted by guanidium thiocyanate method. After first amplification, the product was reamplified by primers labelled with biotin and I-125. The final amplification product was detected by counting the radioactivity after incubation in avidin coated tubes. In 51 samples, the test was repeated for evaluation of reproducibility. This new method was also compared with conventional RT-PCR methods in 34 samples from patients with chronic liver disease. The results were as follows, 1) HCV-RNA was positive in 85(97%)of 88 patients with chronic liver disease, and in 23 (73%) of 30 patients with normal liver function. 2) In comparison with conventional method, HCV-RNA was detected in 32(94%) of 34 patients with new method, whereas in 27(79% ) of the same group with conventional method 3) Repeated test with new method in 52 samples demonstrated 82% of concordant result. In conclusion, new method with biotinylated and radioiodinated primers was more sensitive than conventional method. However, great care must be taken for quality control because there were considerable interassay variation and possibility of false positivity and false negativity.

  13. Quantification of mRNA Levels by Fluorescently Labelled Reverse Transcription Competitive PCR

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    A reproducible,quantitative,non-radioactive method for the analysis of mRNA expression is described.After RNA preparation and cDNA synthesi s,the cDNA was co-amplified with an internal standard in the same PCR system.Th e PCR products containing both targen and internal standard amplificates were el ectrophoresed and detected on an ABI 377 DNA Sequencer.For each sample,β-actin was also quantified by an identical procedure to compensate for relative differ ences between samples in the integrity of the individual RNA samples and for var iations in reverse transcription.Due to the linear relationship between cDNA con tent and PCR product ratio of target cDNA template and competitive standard,a si ngle PCR reaction was sufficient for quantification of a sample.The experimental results showed that the method is a mRNA quantitative RT-PCR method with high sensitivity and good reproducibility.It can be used in large-scale accurate qu antitative analyses of mRNA expression of any gene.

  14. Effect of One-Step and Multi-Steps Polishing System on Enamel Roughness

    Directory of Open Access Journals (Sweden)

    Cynthia Sumali

    2013-07-01

    Full Text Available Normal 0 false false false MicrosoftInternetExplorer4 The final procedures of orthodontic treatment are bracket debonding and cleaning the remaining adhesive. Multi-step polishing system is the most common method used. The disadvantage of that system is long working time, because of the stages that should be done. Therefore, dental material manufacturer make an improvement to the system, to reduce several stages into one stage only. This new system is known as one-step polishing system. Objective: To compare the effect of one-step and multi-step polishing system on enamel roughness after orthodontic bracket debonding. Methods: Randomized control trial was conducted included twenty-eight maxillary premolar randomized into two polishing system; one-step OptraPol (Ivoclar, Vivadent and multi-step AstroPol (Ivoclar, Vivadent. After bracket debonding, the remaining adhesive on each group was cleaned by subjective polishing system for ninety seconds using low speed handpiece. The enamel roughness was subjected to profilometer, registering two roughness parameters (Ra, Rz. Independent t-test was used to analyze the mean score of enamel roughness in each group. Results: There was no significant difference of enamel roughness between one-step and multi-step polishing system (p>0.005. Conclusion: One-step polishing system can produce a similar enamel roughness to multi-step polishing system after bracket debonding and adhesive cleaning.DOI: 10.14693/jdi.v19i3.136

  15. Evaluation of one-step micro polishers for residual resin removal after debonding on fluorosed teeth

    Directory of Open Access Journals (Sweden)

    Padmalatha Challa

    2014-01-01

    Full Text Available Aim and objectives: To evaluate the effectiveness of one step micro polishers for residual resin removal on fluorosed teeth using scanning electron microscope (SEM. Methods and Material: 55 teeth with mild to moderate fluorosis were selected with five teeth as control. Metal brackets were bonded onto 50 teeth which were divided into 5 groups. The finishing and polishing methods which were tested include tungsten carbide burs (TCB, multistep finishing system (Sof-Lex, one step polishers (PoGo and combination of TCB with multistep and one step polishing systems. After resin removal, all the samples were examined under SEM for assessment of the enamel surface. Results: The enamel surface was closest to untouched enamel in samples finished with the PoGo one step polishers followed by Sof-Lex multistep finishing system. However, they took the longest time to finish. TCB required the shortest time for residual resin removal. Conclusions: All polishing systems produce a certain degree of damage to the enamel surface with the smoothest surface being produced by one step polishers on fluorosed teeth.

  16. HIGH ORDER ONE-STEP A-STABLE EXPONENTIALLY FITTED METHODS

    Institute of Scientific and Technical Information of China (English)

    YangFengiian; ChenXinming; LuoYiping

    1999-01-01

    In this paper,the necessary and sutlicient conditions for general one-step methoos to be exponentially fitted at q0∈C are given, A class of multtderivative hybrid one-step methods of order at least s+1 is constructed with s+1 parameters,where s is the order of derivative. The necessary and sufficient conditions for these methods to be A-stable and exponentially fitted is proved, Furthermore,a class of A-stable 2 parameters hybrid one-step methods of order at least 8 are constructed,which use 4th order derivative,These methods are exponentially fitted at q0 if and only its fitted function f(q) satisfies f(q0)= 0, Finally,an A-stable exponentlally fitted method of order 8 is obtained.

  17. Canine distemper virus detection by different methods of One-Step RT-qPCR

    Directory of Open Access Journals (Sweden)

    Claudia de Camargo Tozato

    2016-01-01

    Full Text Available ABSTRACT: Three commercial kits of One-Step RT-qPCR were evaluated for the molecular diagnosis of Canine Distemper Virus. Using the kit that showed better performance, two systems of Real-time RT-PCR (RT-qPCR assays were tested and compared for analytical sensitivity to Canine Distemper Virus RNA detection: a One-Step RT-qPCR (system A and a One-Step RT-qPCR combined with NESTED-qPCR (system B. Limits of detection for both systems were determined using a serial dilution of Canine Distemper Virus synthetic RNA or a positive urine sample. In addition, the same urine sample was tested using samples with prior centrifugation or ultracentrifugation. Commercial kits of One-Step RT-qPCR assays detected canine distemper virus RNA in 10 (100% urine samples from symptomatic animals tested. The One-Step RT-qPCR kit that showed better results was used to evaluate the analytical sensitivity of the A and B systems. Limit of detection using synthetic RNA for the system A was 11 RNA copies µL-1 and 110 RNA copies µl-1 for first round System B. The second round of the NESTED-qPCR for System B had a limit of detection of 11 copies µl-1. Relationship between Ct values and RNA concentration was linear. The RNA extracted from the urine dilutions was detected in dilutions of 10-3 and10-2 by System A and B respectively. Urine centrifugation increased the analytical sensitivity of the test and proved to be useful for routine diagnostics. The One-Step RT-qPCR is a fast, sensitive and specific method for canine distemper routine diagnosis and research projects that require sensitive and quantitative methodology.

  18. Simple One-Step Method to Synthesize Polypyrrole-Indigo Carmine-Silver Nanocomposite

    Directory of Open Access Journals (Sweden)

    Lara Fernandes Loguercio

    2016-01-01

    Full Text Available A nanocomposite of indigo carmine doped polypyrrole/silver nanoparticles was obtained by a one-step electrochemical process. The nanocomposite was characterized by scanning electron microscopy, infrared spectroscopy, ultraviolet-visible-near infrared spectroscopy, and cyclic voltammetry. The simple one-step process allowed the growth of silver nanoparticles during the polymerization of polypyrrole, resulting in films with electrochromic behavior and improved electroactivity. In addition, polypyrrole chains in the nanocomposite were found to present longer conjugation length than pristine polypyrrole films.

  19. The symmetric MSD encoder for one-step adder of ternary optical computer

    Science.gov (United States)

    Kai, Song; LiPing, Yan

    2016-08-01

    The symmetric Modified Signed-Digit (MSD) encoding is important for achieving the one-step MSD adder of Ternary Optical Computer (TOC). The paper described the symmetric MSD encoding algorithm in detail, and developed its truth table which has nine rows and nine columns. According to the truth table, the state table was developed, and the optical-path structure and circuit-implementation scheme of the symmetric MSD encoder (SME) for one-step adder of TOC were proposed. Finally, a series of experiments were designed and performed. The observed results of the experiments showed that the scheme to implement SME was correct, feasible and efficient.

  20. One-step Conversion of Furfural into 2-Methyltetrahydrofuran under Mild Conditions.

    Science.gov (United States)

    Dong, Fang; Zhu, Yulei; Ding, Guoqiang; Cui, Jinglei; Li, Xianqing; Li, Yongwang

    2015-05-11

    One-step direct conversion of biomass-derived furfural to 2-methyltetrahydrofuran was realized under atmospheric pressure over a dual solid catalyst based on two-stage-packed Cu-Pd in a reactor; this is the first report that one-step conversion of furfural resulted in high yield of 2-methyltetrahydrofuran (97.1 %) under atmospheric pressure. This strategy provided a successive hydrogenation process, which avoids high H2 pressure, uses the reactor efficiently, and eliminates the product-separation step. Therefore, it could enhance the overall efficiency as a result of low cost and high yield.

  1. Development of one-step Loop-Mediated Isothermal Amplification (LAMP) for the detection of norovirus in oysters

    Science.gov (United States)

    The aim of this study was to develop a simple and rapid technique for detecting human norovirus (NoV). The loop-mediated isothermal amplification (LAMP) technique was evaluated and found to be sensitive, highly specific, and useful for routine oyster testing. Reverse transcription-LAMP (RT-LAMP) pri...

  2. VP4 and VP7 genotyping by reverse transcription-PCR of human rotavirus in mexican children with acute diarrhea.

    Science.gov (United States)

    Rodríguez Castillo, A; Villa, A V; Ramírez González, J E; Mayén Pimentel, E; Melo Munguía, M; Díaz De Jesús, B; Olivera Díaz, H; García Lozano, H

    2000-10-01

    Dual typing (VP4 and VP7) of rotavirus obtained from 257 Mexican children during three epidemiological seasons was performed by reverse transcription-PCR. The P1G1 genotype was the most prevalent (40%), followed by P1G3 (19%) and P2G2 (16%). Thirty-one specimens (12%) presented mixed infections, while some genotypes were not found. This is the first dual typing of isolates from diarrhea cases in Mexico.

  3. Development and Validation of a Multiplex Reverse Transcription PCR Assay for Simultaneous Detection of Three Papaya Viruses

    OpenAIRE

    Tuo, Decai; Shen, Wentao; Yang, Yong; Yan, Pu; Li, Xiaoying; Zhou, Peng

    2014-01-01

    Papaya ringspot virus (PRSV), Papaya leaf distortion mosaic virus (PLDMV), and Papaya mosaic virus (PapMV) produce similar symptoms in papaya. Each threatens commercial production of papaya on Hainan Island, China. In this study, a multiplex reverse transcription PCR assay was developed to detect simultaneously these three viruses by screening combinations of mixed primer pairs and optimizing the multiplex RT-PCR reaction conditions. A mixture of three specific primer pairs was used to amplif...

  4. One-step Multiplex Transgenesis via Sleeping Beauty Transposition in Cattle.

    Science.gov (United States)

    Garrels, Wiebke; Talluri, Thirumala R; Apfelbaum, Ronja; Carratalá, Yanet P; Bosch, Pablo; Pötzsch, Kerstin; Grueso, Esther; Ivics, Zoltán; Kues, Wilfried A

    2016-02-24

    Genetically modified cattle are important for developing new biomedical models and for an improved understanding of the pathophysiology of zoonotic diseases. However, genome editing and genetic engineering based on somatic cell nuclear transfer suffer from a low overall efficiency. Here, we established a highly efficient one-step multiplex gene transfer system into the bovine genome.

  5. Atomic GHZ States Prepared in Two Directly Coupled Cavities with Virtual Excitations in One Step

    Institute of Scientific and Technical Information of China (English)

    杨榕灿; 黄志平; 郭强; 张鹏飞; 钟纯勇; 张天才

    2011-01-01

    A scheme for one-step preparation of atomic GHZ states in two directly coupled cavities via virtual excitations is proposed. In the whole procedure, the information is carried only in two ground states of A-type atoms, while the excited states of atoms and cavity modes are virtually excited, leading the system to be insensitive to atomic spontaneous emission and photon loss.

  6. Fast-ion energy resolution by one-step reaction gamma-ray spectrometry

    DEFF Research Database (Denmark)

    Salewski, Mirko; Nocente, M.; Gorini, G.

    2016-01-01

    The spectral broadening of γ-rays from fusion plasmas can be measured in high-resolution gamma-ray spectrometry (GRS). We derive weight functions that determine the observable velocity space and quantify the velocity-space sensitivity of one-step reaction high-resolution GRS measurements in magne...

  7. One-step synthesis of silver nanoparticle-filled Nylon 6 nanofibers and their antibacterial properties

    Science.gov (United States)

    A novel and facile one-step approach to in situ synthesize silver nanoparticle-filled nylon 6 nanofibers by electrospinning is reported. The method does not need post-treatments and can be carried out at ambient conditions without using additional chemicals. It employs the electrospinning solvent as...

  8. Noether's theorem and one-step corrections method for holonomic system

    Institute of Scientific and Technical Information of China (English)

    Shang Mei; Chen Xiang-Wei

    2006-01-01

    In this paper, a new computational method for improving the accuracy of numerically computed solutions is introduced. The computational method is based on the one-step method and conserved quantities of holonomic systems are considered as kinematical constraints in this method.

  9. One-step synthesis of three-dimensional Pd polyhedron networks with enhanced electrocatalytic performance.

    Science.gov (United States)

    Xu, You; Xu, Rui; Cui, Jianhua; Liu, Yang; Zhang, Bin

    2012-04-21

    Three-dimensional Pd polyhedron networks (Pd PNs) have been fabricated for the first time through a one-step, Cu(2+)-assisted, solution-chemical approach. These as-prepared 3D Pd PNs exhibit high stability and remarkably improved electrocatalytic activity toward formic acid oxidation over commercially available Pd black. This journal is © The Royal Society of Chemistry 2012

  10. Constraint Stabilization and One-Step Correction Method for Dynamical Systems

    Institute of Scientific and Technical Information of China (English)

    SHANG Mei; CHEN Xiang-wei; MEI Feng-xiang

    2007-01-01

    A computational method of constraint stabilization and correction is introduced.The method is based on the Baumgart's one-step method.Constraint conditions are addressed to stabilize and correct the solution.Two examples are given to illustrate the results of the method.

  11. One-step polymer surface modification for minimizing drug, protein, and DNA adsorption in microanalytical systems

    DEFF Research Database (Denmark)

    Larsen, Esben Kjær Unmack; Larsen, Niels Bent

    2013-01-01

    The non-specific adsorption of dissolved analytes strongly reduces the sensitivity and reliability in polymer microanalytical systems. Here, a one-step aqueous phase procedure modifies polymer material surfaces to strongly reduce their non-specific adsorption of a broad range of organic analytes ...... systems, including polystyrene (PS), cyclic olefin copolymer (COC), liquid crystalline polymer (LCP), and polyimide (PI)....

  12. Two Steps Forward, One Step Backward: Must This Be the Future of Diversity?

    Science.gov (United States)

    Butler, Johnnella E.

    2013-01-01

    Johnnella Butler writes here that the title of this article "Two Steps Forward, One Step Backward," expresses the "wicked problem" of diversity as a concrete goal in higher education. The concept of the "wicked problem," is a term coined in the late 1960s by social planners. Consulting Wikipedia, as so many of our…

  13. Development of a one step strip test for the detection of sulfadimidine residues

    NARCIS (Netherlands)

    Verheijen, R.; Stouten, P.; Cazemier, G.; Haasnoot, W.

    1998-01-01

    The one step strip test described is a competitive immunoassay in which the detector reagent consists of colloidal gold particles coated with affinity purified polyclonal anti-sulfadimidine (SDD) antibodies. The capture reagent in the assay is an SDD–ovalbumin conjugate which is immobilised on the l

  14. One step to generate quantum controlled phase-shift gate using a trapped ion

    Institute of Scientific and Technical Information of China (English)

    Zhang Shi-Jun; Ma Chi; Zhang Wen-Hai; Ye Liu

    2008-01-01

    This paper presents a very simple scheme for generating quantum controlled phase-shift gate with only one step by using the two vibrational modes of a trapped ion as the two qubits.The scheme couples two vibration degrees of freedom coupled with a suitable chosen laser excitation via the ionic states.

  15. One-step isolation of plasma membrane proteins using magnetic beads with immobilized concanavalin A

    DEFF Research Database (Denmark)

    Lee, Yu-Chen; Block, Gregory; Chen, Huiwen;

    2008-01-01

    We have developed a simple method for isolating and purifying plasma membrane proteins from various cell types. This one-step affinity-chromatography method uses the property of the lectin concanavalin A (ConA) and the technique of magnetic bead separation to obtain highly purified plasma membran...

  16. High fat diet-induced changes of mouse hepatic transcription and enhancer activity can be reversed by subsequent weight loss

    DEFF Research Database (Denmark)

    Siersbæk, Majken; Varticovski, Lyuba; Yang, Shutong

    2017-01-01

    of chow, to identify HFD-mediated changes to the hepatic transcriptional program that may persist after weight loss. Mice fed a HFD displayed increased fasting insulin levels, hepatosteatosis and major changes in hepatic gene transcription associated with modulation of H3K27Ac at enhancers...... fully restored to normal levels. Moreover, HFD-regulated H3K27Ac and mRNA levels returned to similar levels as control mice. These data demonstrates that the transcription regulatory landscape in the liver induced by HFD is highly dynamic and can be reversed by weight loss. This provides hope...... for efficient treatment of early obesity-associated changes to hepatic complications by simple weight loss intervention without persistent reprograming of the liver transcriptome....

  17. Interphase fluorescence in situ hybridization and reverse transcription polymerase chain reaction as a diagnostic aid for synovial sarcoma.

    Science.gov (United States)

    Shipley, J; Crew, J; Birdsall, S; Gill, S; Clark, J; Fisher, C; Kelsey, A; Nojima, T; Sonobe, H; Cooper, C; Gusterson, B

    1996-02-01

    Identification of the t(X;18)(p11.2;q11.2) that is associated with a high proportion of synovial sarcoma can be a useful diagnostic aid. The translocation results in fusion of the SYT gene on chromosome 18 to either the SSX1 or the SSX2 gene, two homologous genes within Xp11.2. Two-color interphase fluorescence in situ hybridization and reverse transcription polymerase chain reaction were assessed as approaches to identify the rearrangement in well characterized cases. The presence of the translocation, and the specific chromosome X gene disrupted, were inferred from the configuration of signals from chromosome-specific centromere probes, paints, and markers flanking each gene in preparations of interphase nuclei. Rearrangement was found in two cell lines and eight of nine tumor samples, including analysis of five touch imprints. This was consistent with cytogenetic data in four cases and reverse transcription polymerase chain reaction analysis using primers known to amplify both SYT-SSX1 and SYT-SSX2 transcripts. The transcripts were distinguished by restriction with LspI and SmaI. Contrary to previous suggestions, there was no obvious correlation between histological subtype and involvement of the SSX1 or SSX2 gene. These approaches could also be applied to the identification of tumor-free margins and metastatic disease.

  18. Efficient in vitro inhibition of HIV-1 gag reverse transcription by peptide nucleic acid (PNA) at minimal ratios of PNA/RNA

    DEFF Research Database (Denmark)

    Koppelhus, Uffe; Zachar, Vladimir; Nielsen, P.E.;

    1997-01-01

    We have tested the inhibitory potential of peptide nucleic acid (PNA) on in vitro reverse transcription of the HIV-1 gag gene. PNA was designed to target different regions of the HIV-1 gag gene and the effect on reverse transcription by HIV-1, MMLV and AMV reverse transcriptases (RTs) was investi......We have tested the inhibitory potential of peptide nucleic acid (PNA) on in vitro reverse transcription of the HIV-1 gag gene. PNA was designed to target different regions of the HIV-1 gag gene and the effect on reverse transcription by HIV-1, MMLV and AMV reverse transcriptases (RTs......) was investigated. We found that a bis-PNA (parallel antisense 10mer linked to antiparallel antisense 10mer) was superior to both the parallel antisense 10mer and antiparallel antisense 10mer in inhibiting reverse transcription of the gene, thus indicating triplex formation at the target sequence. A complete arrest...... that would indicate PNA-mediated RNase H activation of the tested RTs. In conclusion, PNA appears to have a potential to become a specific and efficient inhibitor of reverse transcription in vivo , provided sufficient intracellular levels are achievable....

  19. A modified reverse one-hybrid screen identifies transcriptional activation in Phyochrome-Interacting Factor 3

    Science.gov (United States)

    Transcriptional activation domains (TAD) are difficult to predict and identify, since they are not conserved and have little consensus. Here, we describe a yeast-based screening method that is able to identify individual amino acid residues involved in transcriptional activation in a high throughput...

  20. One-Step Dynamic Classifier Ensemble Model for Customer Value Segmentation with Missing Values

    Directory of Open Access Journals (Sweden)

    Jin Xiao

    2014-01-01

    Full Text Available Scientific customer value segmentation (CVS is the base of efficient customer relationship management, and customer credit scoring, fraud detection, and churn prediction all belong to CVS. In real CVS, the customer data usually include lots of missing values, which may affect the performance of CVS model greatly. This study proposes a one-step dynamic classifier ensemble model for missing values (ODCEM model. On the one hand, ODCEM integrates the preprocess of missing values and the classification modeling into one step; on the other hand, it utilizes multiple classifiers ensemble technology in constructing the classification models. The empirical results in credit scoring dataset “German” from UCI and the real customer churn prediction dataset “China churn” show that the ODCEM outperforms four commonly used “two-step” models and the ensemble based model LMF and can provide better decision support for market managers.

  1. THE CRITICAL LOAD PARAMETER OF A TIMOSHENKO BEAM WITH ONE-STEP CHANGE IN CROSS SECTION

    Directory of Open Access Journals (Sweden)

    Goran Janevski

    2014-12-01

    Full Text Available The paper analyzes the transverse vibration of a Timoshenko beam with one-step change in cross-section when subjected to an axial force. The axial force is equal in both of the beam portions. Three types of beam which occur commonly in engineering application are considered. The frequency equation of the Timoshenko beam with one-step change in cross-section is expressed as the fourth order determinant equated to zero. The critical compressive axial force is expressed as a function of the critical load parameter which is tabulated for four classical boundary conditions. Apart from the results presented in Tables, the paper also provides calculated values of the critical load parameter for other values of system parameters along with the graphic representation of their dependence on the step position parameter.

  2. One Step Combustion Synthesis Of YAG:Ce Phosphor For Solid State Lighting

    Science.gov (United States)

    Yadav, Pooja; Gupta, K. Vijay Kumar; Muley, Aarti; Joshi, C. P.; Moharil, S. V.

    2011-10-01

    YAG:Ce is an important phosphor having applications in various fields ranging from solid state lighting to scintillation detectors. YAG phosphors doped with activators are mainly synthesized by solid state reaction techniques that require high sintering temperatures (above 1500°C) to eliminate YAM and YAP phases. Though several soft chemical routes have been explored for synthesis of YAG, most of these methods are complex and phase pure materials are not obtained in one step, but prolonged annealing at temperatures around 1000 C or above becomes necessary. One step combustion synthesis of YAG:Ce3+ and related phosphors carried out at 500 C furnace temperature is reported here. Activation with Ce3+ could be achieved during the synthesis without taking recourse to any post-combustion thermal treatment. LEDs prepared from the combustion synthesized YAG:Ce3+, exhibited properties comparable to those produced from the commercial phosphor.

  3. One-Step Implementation of Single Spin Measurement in Spin Star Network

    Institute of Scientific and Technical Information of China (English)

    DENG Hong-Liang; FANG Xi-Ming

    2008-01-01

    We present an efficient one-step scheme for a single spin measurement based on nuclear magnetic resonance (NMR)techniques.This scheme considerably reduces the time of operation using a spin star network where a target spin and an ancillary spin are coupled to a ring of N spins.As opposed to the proposal in [Phys.Rev.Lett.97(2006)100501]using a cubic lattice crystal to achieve a cubic speedup,the distinct advantage of this scheme is that under ideal conditions it requires the application of only one step to create a system of N correlated spins.In the process of single spin measurement,the total macroscopic magnetization,the individual magnetization and the transfer fideity are calculated analytically as simple cosine functions of time and the amplitude of irradiation.

  4. One-step purification of metallothionein extracted from two different sources.

    Science.gov (United States)

    Honda, Rubens T; Araújo, Roziete Mendes; Horta, Bruno Brasil; Val, Adalberto L; Demasi, Marilene

    2005-06-25

    We describe a one-step purification of hepatic metallothionein from the Amazon fish Colossoma macropomum injected with cadmium and from the copper-loaded metallothionein from the yeast Saccharomyces cerevisiae, performed by affinity chromatography through metal-chelating columns. Yeast metallothionein was purified from Cu2+-loaded resin and eluted by a continuous EDTA gradient whereas hepatic metallothionein extracted from fishes was purified by Ni2+-loaded resin and eluted by a continuous imidazol gradient. Purified metallothioneins were evaluated by SDS-PAGE and characterized by UV spectra of the apo- and Cd2+-loaded protein. This method allowed high purity and yield as well as rapid one-step extraction of both metal-loaded and apoprotein.

  5. One-step solution combustion synthesis of pure Ni nanopowders with enhanced coercivity: The fuel effect

    Science.gov (United States)

    Khort, Alexander; Podbolotov, Kirill; Serrano-García, Raquel; Gun'ko, Yurii K.

    2017-09-01

    In this paper, we report a new modified one-step combustion synthesis technique for production of Ni metal nanoparticles. The main unique feature of our approach is the use of microwave assisted foam preparation. Also, the effect of different types of fuels (urea, citric acid, glycine and hexamethylenetetramine) on the combustion process and characteristics of resultant solid products were investigated. It is observed that the combination of microwave assisted foam preparation and using of hexamethylenetetramine as a fuel allows producing pure ferromagnetic Ni metal nanoparticles with enhanced coercivity (78 Oe) and high value of saturation magnetization (52 emu/g) by one-step solution combustion synthesis under normal air atmosphere without any post-reduction processing.

  6. High fat diet-induced changes of mouse hepatic transcription and enhancer activity can be reversed by subsequent weight loss.

    Science.gov (United States)

    Siersbæk, Majken; Varticovski, Lyuba; Yang, Shutong; Baek, Songjoon; Nielsen, Ronni; Mandrup, Susanne; Hager, Gordon L; Chung, Jay H; Grøntved, Lars

    2017-01-10

    Epigenetic factors have been suggested to play an important role in metabolic memory by trapping and maintaining initial metabolic changes within the transcriptional regulatory machinery. In this study we fed mice a high fat diet (HFD) for seven weeks followed by additional five weeks of chow, to identify HFD-mediated changes to the hepatic transcriptional program that may persist after weight loss. Mice fed a HFD displayed increased fasting insulin levels, hepatosteatosis and major changes in hepatic gene transcription associated with modulation of H3K27Ac at enhancers, but no significant changes in chromatin accessibility, indicating that HFD-regulated gene transcription is primarily controlled by modulating the activity of pre-established enhancers. After return to the same body weight as chow fed control mice, the fasting insulin, glucose, and hepatic triglyceride levels were fully restored to normal levels. Moreover, HFD-regulated H3K27Ac and mRNA levels returned to similar levels as control mice. These data demonstrates that the transcription regulatory landscape in the liver induced by HFD is highly dynamic and can be reversed by weight loss. This provides hope for efficient treatment of early obesity-associated changes to hepatic complications by simple weight loss intervention without persistent reprograming of the liver transcriptome.

  7. Fast-ion energy resolution by one-step reaction gamma-ray spectrometry

    DEFF Research Database (Denmark)

    Salewski, Mirko; Nocente, M.; Gorini, G.;

    2016-01-01

    The spectral broadening of γ-rays from fusion plasmas can be measured in high-resolution gamma-ray spectrometry (GRS). We derive weight functions that determine the observable velocity space and quantify the velocity-space sensitivity of one-step reaction high-resolution GRS measurements in magne...... general formalism using reactions with and without intrinsic broadening of the γ-rays for the GRS diagnostic at JET....

  8. Ethanol Manufacture through One-step Cellulose Liquefaction Developed by Zhongren Bioenergy Company

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    @@ The pilot scale tests of one-step direct liquefaction of cel-lulose biomass developed by a Sino-US joint venture, the Huaibei Zhongren Bioenergy Technical Development Company, Ltd. in Anhui province, have made great success. This method aiming to produce fuel and chemical feedstocks from cellulose biomass requires mild reaction conditions and all organic substances contained in the cellulose biom-ass can be completely converted without losses (without carbonization and gasification).

  9. Explicit One-Step P-Stable Methods for Second Order Periodic Initial Value Problems

    Institute of Scientific and Technical Information of China (English)

    Qinghong Li; Yongzhong Song

    2006-01-01

    In this paper, we present an explicit one-step method for solving periodic initial value problems of second order ordinary differential equations. The method is P-stable, and of first algebraic order and high phase-lag order. To improve the algebraic order, we give a composition second order scheme with the proposed method and its adjoint. We report some numerical results to illustrate the efficiency of our methods.

  10. Profilometry with Grating Projection Based on One-step Phase Shift

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    An optical technique for 3-D shape measurement is presented. This technique, based on a deformed projected grating pattern which carries 3-D information of the measured object, can automatically and accurately obtain the phase map of a measured object by using one-step phase shift algorithm.In comparison with traditional phase-shift technique, the technique is much faster, with the equivalent accuracy. Only one frame image is sufficient for measuring. Experimental result of typical object is presented.

  11. Genomic prediction for Nordic Red Cattle using one-step and selection index blending

    DEFF Research Database (Denmark)

    Guosheng, Su; Madsen, Per; Nielsen, Ulrik Sander

    2012-01-01

    This study investigated the accuracy of direct genomic breeding values (DGV) using a genomic BLUP model, genomic enhanced breeding values (GEBV) using a one-step blending approach, and GEBV using a selection index blending approach for 15 traits of Nordic Red Cattle. The data comprised 6,631 bull......-step blending approach is a good alternative to predict GEBV in practical genetic evaluation program....

  12. Simple Scheme for Directly Measuring Concurrence of Two-Qubit Pure States in One Step

    Institute of Scientific and Technical Information of China (English)

    YANG Rong-Can; LIN Xiu; HUANG Zhi-Ping; LI Hong-Cai

    2009-01-01

    In the present work, a simple scheme for the direct measurement of the concurrence of two-qubit pure states is proposed.The scheme is based on trapped ions and only needs one step when the two identical pure states are given.The vibrational mode in our proposal is only virtually excited, which is important in view of decoherence.Furthermore, the scheme is feasible based on current technologies.

  13. One-Step Dynamic Classifier Ensemble Model for Customer Value Segmentation with Missing Values

    OpenAIRE

    Jin Xiao; Bing Zhu; Geer Teng; Changzheng He; Dunhu Liu

    2014-01-01

    Scientific customer value segmentation (CVS) is the base of efficient customer relationship management, and customer credit scoring, fraud detection, and churn prediction all belong to CVS. In real CVS, the customer data usually include lots of missing values, which may affect the performance of CVS model greatly. This study proposes a one-step dynamic classifier ensemble model for missing values (ODCEM) model. On the one hand, ODCEM integrates the preprocess of missing values and the classif...

  14. One-step synthesis of silver nanoparticles, nanorods, and nanowires on the surface of DNA network.

    Science.gov (United States)

    Wei, Gang; Zhou, Hualan; Liu, Zhiguo; Song, Yonghai; Wang, Li; Sun, Lanlan; Li, Zhuang

    2005-05-12

    Here, we describe a one-step synthesis of silver nanoparticles, nanorods, and nanowires on DNA network surface in the absence of surfactant. Silver ions were first adsorbed onto the DNA network and then reduced in sodium borohydride solution. Silver nanoparticles, nanorods, and nanowires were formed by controlling the size of pores of the DNA network. The diameter of the silver nanoparticles and the aspect ratio of the silver nanorods and nanowires can be controlled by adjusting the DNA concentration and reduction time.

  15. One-step synthesis of 1-chloro-3-arylacetone derivatives from arylacetic acids.

    Science.gov (United States)

    Zacuto, Michael J; Dunn, Robert F; Figus, Margaret

    2014-09-19

    A practical one-step method has been developed to prepare α-chloroketones from readily available, inexpensive phenylacetic acid derivatives. The method utilizes the unique reactivity of an intermediate Mg-enolate dianion, which displays selectivity for the carbonyl carbon of chloromethyl carbonyl electrophiles. Decarboxylation of the intermediate occurs spontaneously during the reaction quench. The utility of the reaction products has been demonstrated through the total synthesis of the natural product cimiracemate B.

  16. One Step Preparation of Controlled Drug Release Systems in Supercritical Carbon Dioxide

    Institute of Scientific and Technical Information of China (English)

    CAO Liqin; WANG Chengwei; CHEN Liuping

    2009-01-01

    Drug delivery systems based on copolymers of N-isopropylacrylamide were fn-st prepared by a one step method, in which supercritical carbon dioxide was simultaneously used as a polymerization medium and an impregnation agent. The obtained microspheres were characterized by scanning electronic microscopy (SEM), differential scan-ning calorimetry (DSC), transmission electron microscopy (TEM) and X-ray diffraction (XRD). The release effect of the in situ prepared microgels impregnated with ibuprofen was presented through in vitro release simulation.

  17. Evaluating formability of LCP plate for sacral fractures with one step inverse forming finite element analysis.

    Science.gov (United States)

    Li, Xiaoda; Zhang, Xiangkui; Hu, Ping; Liu, Weijie; Shen, Guozhe; Zhan, Xianghui

    2015-01-01

    The locking compression plate fixation treatment for the unstable sacral fractures is simple and effective, with less trauma and complications. Some locking compression plate parts have been made of high-strength Plate manufactured by hot stamping process since the demand for lightweight biomedical materials. Finite Element (FE) method of One-Step inverse forming based on deformation theory is the tool to evaluate the formability of locking compression plate panel quickly in initial design for reducing costs and development cycle of Plate. But current one-step inverse forming methods are all suitable for cold stamping, not hot-stamping. This paper proposed one-step inverse forming method and workflow for hot-stamping of locking compression Plate. And the B pillar of a sacral bone was simulated and its computing result was compared with experimental value. The result shows that the proposed method in this paper can quickly evaluate high temperature formability of high-strength Plate. And the method is proposed to be used in initial design.

  18. One-step graphene coating of heteroepitaxial GaN films.

    Science.gov (United States)

    Choi, Jae-Kyung; Huh, Jae-Hoon; Kim, Sung-Dae; Moon, Daeyoung; Yoon, Duhee; Joo, Kisu; Kwak, Jinsung; Chu, Jae Hwan; Kim, Sung Youb; Park, Kibog; Kim, Young-Woon; Yoon, Euijoon; Cheong, Hyeonsik; Kwon, Soon-Yong

    2012-11-02

    Today, state-of-the-art III-Ns technology has been focused on the growth of c-plane nitrides by metal-organic chemical vapor deposition (MOCVD) using a conventional two-step growth process. Here we show that the use of graphene as a coating layer allows the one-step growth of heteroepitaxial GaN films on sapphire in a MOCVD reactor, simplifying the GaN growth process. It is found that the graphene coating improves the wetting between GaN and sapphire, and, with as little as ~0.6 nm of graphene coating, the overgrown GaN layer on sapphire becomes continuous and flat. With increasing thickness of the graphene coating, the structural and optical properties of one-step grown GaN films gradually transition towards those of GaN films grown by a conventional two-step growth method. The InGaN/GaN multiple quantum well structure grown on a GaN/graphene/sapphire heterosystem shows a high internal quantum efficiency, allowing the use of one-step grown GaN films as 'pseudo-substrates' in optoelectronic devices. The introduction of graphene as a coating layer provides an atomic playground for metal adatoms and simplifies the III-Ns growth process, making it potentially very useful as a means to grow other heteroepitaxial films on arbitrary substrates with lattice and thermal mismatch.

  19. A one step real time PCR method for the quantification of hepatitis delta virus RNA using an external armored RNA standard and intrinsic internal control.

    Science.gov (United States)

    Karataylı, Ersin; Altunoğlu, Yasemin Çelik; Karataylı, Senem Ceren; Alagöz, S Gökçe K; Cınar, Kubilay; Yalçın, Kendal; Idilman, Ramazan; Yurdaydın, Cihan; Bozdayı, A Mithat

    2014-05-01

    Hepatitis delta virus (HDV) RNA viral load measurement is critical in diagnosis and monitoring the response to antiviral treatment. Our aim is to design a real time PCR method for accurate quantitation of HDV RNA in clinical specimens using an armored RNA as external standard, and an intrinsic internal control. A plasmid bearing delta antigen region of genotype I HDV genome was used to develop an armored RNA. Serial dilutions of the armored HDV RNA standard with 10(12)copy/mL were used as standards for quantitation. A primer-probe set derived from HDAg region was used in one step EZ RT PCR kit chemistry which uses rTth enzyme allowing reverse transcription and polymerization in the same tube. The kit also uses the advantage of uracil-N-glycosylase (UNG) enzyme treatment to prevent PCR contamination. The established assay has a dynamic range of 10(2)-10(11)copy/mL with a PCR efficiency of 96.9%. Detection limit was 858±32copy/mL with 95% confidence interval. Intra- and inter-assay variabilities were low for high, medium and low levels of viremia. Incorporation of freely circulating GAPDH in serum into the assay as an intrinsic internal control prevented false negative results and failures in PCR amplifications due to inhibitors, inefficient extraction procedures or enzymatic reactions. In conclusion, this study defines a novel assay for sensitive and reliable quantification of HDV RNA using an armored HDV RNA as a standard and GAPDH in plasma or serum as an intrinsic internal control in a single tube. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Reverse Genetic Analysis of Transcription FactorOsHox9, a Member of Homeobox Family, in Rice

    Institute of Scientific and Technical Information of China (English)

    AI Li-ping; SHEN Ao; GAO Zhi-chao; LI Zheng-long; SUN Qiong-lin; LI Ying-ying; LUAN Wei-jiang

    2014-01-01

    Homeobox transcription factors participate in the growth and development of plants by regulating cell differentiation, morphogenesis and environmental signal response. To reveal the functions of these transcription factors in rice, we constructed the RNAi vectors ofOsHox9, a member of homeobox family, and analyzed the function ofOsHox9 using reverse genetics. The plant height and tillering number of RNAi transgenic plants decreased compared with those of wild-type plants. Reverse transcription-polymerase chain reaction analysis showed thatOsHox9 expression reduced in the transgenic plants with phenotypic variance, whereas that in the transgenic plants without phenotypic variance was similar to that in the wild-type plants. This result suggests that the phenotypes of the transgenic plants were caused by RNAi effects. The tissue-specificity ofOsHox9 expression indicated that it was expressed in different organs, with high expression in stem apical meristem and young panicles. Subcelular location ofOsHox9 demonstrated that it was localized on the cell membrane.

  1. Reverse engineering a mouse embryonic stem cell-specific transcriptional network reveals a new modulator of neuronal differentiation.

    Science.gov (United States)

    De Cegli, Rossella; Iacobacci, Simona; Flore, Gemma; Gambardella, Gennaro; Mao, Lei; Cutillo, Luisa; Lauria, Mario; Klose, Joachim; Illingworth, Elizabeth; Banfi, Sandro; di Bernardo, Diego

    2013-01-01

    Gene expression profiles can be used to infer previously unknown transcriptional regulatory interaction among thousands of genes, via systems biology 'reverse engineering' approaches. We 'reverse engineered' an embryonic stem (ES)-specific transcriptional network from 171 gene expression profiles, measured in ES cells, to identify master regulators of gene expression ('hubs'). We discovered that E130012A19Rik (E13), highly expressed in mouse ES cells as compared with differentiated cells, was a central 'hub' of the network. We demonstrated that E13 is a protein-coding gene implicated in regulating the commitment towards the different neuronal subtypes and glia cells. The overexpression and knock-down of E13 in ES cell lines, undergoing differentiation into neurons and glia cells, caused a strong up-regulation of the glutamatergic neurons marker Vglut2 and a strong down-regulation of the GABAergic neurons marker GAD65 and of the radial glia marker Blbp. We confirmed E13 expression in the cerebral cortex of adult mice and during development. By immuno-based affinity purification, we characterized protein partners of E13, involved in the Polycomb complex. Our results suggest a role of E13 in regulating the division between glutamatergic projection neurons and GABAergic interneurons and glia cells possibly by epigenetic-mediated transcriptional regulation.

  2. Efficient reverse transcription using locked nucleic acid nucleotides towards the evolution of nuclease resistant RNA aptamers

    DEFF Research Database (Denmark)

    Crouzier, Lucile; Dubois, Camille; Edwards, Stacey L

    2012-01-01

    We found that SuperScript® III Reverse Transcriptase is an efficient enzyme for the recognition of LNA nucleotides, making it a prime candidate to be used in de novo selection of LNA containing RNA aptamers....

  3. One-step hydrothermal synthesis of three-dimensional porous graphene aerogels/sulfur nanocrystals for lithium–sulfur batteries

    Energy Technology Data Exchange (ETDEWEB)

    Jiang, Yong; Lu, Mengna; Ling, Xuetao; Jiao, Zheng; Chen, Lingli; Chen, Lu [School of Environmental and Chemical Engineering, Shanghai University, Shanghai 200444 (China); Hu, Pengfei [Instrumental Analysis and Research Center, Shanghai University, Shanghai 200444 (China); Zhao, Bing, E-mail: bzhao@shu.edu.cn [School of Environmental and Chemical Engineering, Shanghai University, Shanghai 200444 (China)

    2015-10-05

    Highlights: • 3D porous GA/S nanocrystals are prepared by a one-step hydrothermal method. • The structure is affected by hydrothermal temperature and liquid sulfur’s viscosity. • The hybrid delivers a capacity of 716.2 mA h g{sup −1} after 50 cycles at 100 mA g{sup −1}. • The nanosized S, strong adsorbability and intimate contact of GNS are main factors. - Abstract: Lithium–sulfur (Li–S) batteries are receiving significant attention as a new energy source because of its high theoretical capacity and specific energy. However, the low sulfur loading and large particles (usually in submicron dimension) in the cathode greatly offset its advantage in high energy density and lead to the instability of the cathode and rapid capacity decay. Herein, we introduce a one-step hydrothermal synthesis of three-dimensional porous graphene aerogels/sulfur nanocrystals to suppress the rapid fading of sulfur electrode. It is found that the hydrothermal temperature and viscosity of liquid sulfur have significant effects on particle size and loading mass of sulfur nanocrystals, graphitization degree of graphene and chemical bonding between sulfur and oxygen-containing groups of graphene. The hybrid could deliver a specific capacity of 716.2 mA h g{sup −1} after 50 cycles at a current density of 100 mA g{sup −1} and reversible capacity of 517.9 mA h g{sup −1} at 1 A g{sup −1}. The performance we demonstrate herein suggests that Li–S battery may provide an opportunity for development of rechargeable battery systems.

  4. Leptin upregulates telomerase activity and transcription of human telomerase reverse transcriptase in MCF-7 breast cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Ren, He, E-mail: herenrh@yahoo.com.cn [Key Laboratory of Breast Cancer Prevention and Therapy, Tianjin Medical University, Ministry of Education, Tianjin Medical University Cancer Hospital, Tianjin (China); Zhao, Tiansuo; Wang, Xiuchao; Gao, Chuntao; Wang, Jian; Yu, Ming [Key Laboratory of Breast Cancer Prevention and Therapy, Tianjin Medical University, Ministry of Education, Tianjin Medical University Cancer Hospital, Tianjin (China); Hao, Jihui, E-mail: jihuihao@yahoo.com [Key Laboratory of Breast Cancer Prevention and Therapy, Tianjin Medical University, Ministry of Education, Tianjin Medical University Cancer Hospital, Tianjin (China)

    2010-03-26

    The aim was to analyze the mechanism of leptin-induced activity of telomerase in MCF-7 breast cancer cells. We found that leptin activated telomerase in a dose-dependent manner; leptin upregulated the expression of Human Telomerase Reverse Transcriptase (hTERT) at mRNA and protein levels; blockade of signal transducer and activator of transcription 3 (STAT3) phosphorylation significantly counteracted leptin-induced hTERT transcription and protein expression; chromatin immunoprecipitation analysis showed that leptin enhanced the binding of STAT3 to the hTERT promoter. This study uncovers a new mechanism of the proliferative effect of leptin on breast cancer cells and provides a new explanation of obesity-related breast cancer.

  5. Reverse engineering of TLX oncogenic transcriptional networks identifies RUNX1 as tumor suppressor in T-ALL.

    Science.gov (United States)

    Della Gatta, Giusy; Palomero, Teresa; Perez-Garcia, Arianne; Ambesi-Impiombato, Alberto; Bansal, Mukesh; Carpenter, Zachary W; De Keersmaecker, Kim; Sole, Xavier; Xu, Luyao; Paietta, Elisabeth; Racevskis, Janis; Wiernik, Peter H; Rowe, Jacob M; Meijerink, Jules P; Califano, Andrea; Ferrando, Adolfo A

    2012-02-26

    The TLX1 and TLX3 transcription factor oncogenes have a key role in the pathogenesis of T cell acute lymphoblastic leukemia (T-ALL). Here we used reverse engineering of global transcriptional networks to decipher the oncogenic regulatory circuit controlled by TLX1 and TLX3. This systems biology analysis defined T cell leukemia homeobox 1 (TLX1) and TLX3 as master regulators of an oncogenic transcriptional circuit governing T-ALL. Notably, a network structure analysis of this hierarchical network identified RUNX1 as a key mediator of the T-ALL induced by TLX1 and TLX3 and predicted a tumor-suppressor role for RUNX1 in T cell transformation. Consistent with these results, we identified recurrent somatic loss-of-function mutations in RUNX1 in human T-ALL. Overall, these results place TLX1 and TLX3 at the top of an oncogenic transcriptional network controlling leukemia development, show the power of network analyses to identify key elements in the regulatory circuits governing human cancer and identify RUNX1 as a tumor-suppressor gene in T-ALL.

  6. Real-time reverse transcription-polymerase chain reaction assays for identification of wild poliovirus 1 & 3

    OpenAIRE

    Sharma, Deepa K.; Nalavade, Uma P.; Deshpande, Jagadish M.

    2015-01-01

    Background & objectives: The poliovirus serotype identification and intratypic differentiation by real-time reverse transcription-polymerase chain reaction (rRT-PCR) assay is suitable for serotype mixtures but not for intratypic mixtures of wild and vaccine poliovirus strains. This study was undertaken to develop wild poliovirus 1 and 3 (WPV1 and WPV3) specific rRT-PCR assays for use. Methods: Specific primers and probes for rRT-PCR were designed based on VP1 sequences of WPV1 and WPV3 isolat...

  7. Target-cell-derived tRNA-like primers for reverse transcription support retroviral infection at low efficiency

    DEFF Research Database (Denmark)

    Schmitz, Alexander; Lund, Anders H; Hansen, Anette C

    2002-01-01

    by cases of correction of single mismatches between Akv-MLV vectors and complementary tRNA primers toward the primer sequence in the integrated vector. Thus, target-cell-derived tRNA-like primers are able to initiate first-strand cDNA synthesis and plus-strand transfer leading to a complete provirus......RNA primers derived from the target cell can sustain reverse transcription during murine leukemia virus (MLV) infection. Transduction efficiencies were 4-5 orders of magnitude below those of comparable producer-cell complementations. However, successful usage of a target-cell-derived tRNA primer was proven...

  8. Use of Existing Diagnostic Reverse-Transcription Polymerase Chain Reaction Assays for Detection of Ebola Virus RNA in Semen.

    Science.gov (United States)

    Pettitt, James; Higgs, Elizabeth S; Adams, Rick D; Jahrling, Peter B; Hensley, Lisa E

    2016-04-15

    Sexual transmission of Ebola virus in Liberia has now been documented and associated with new clusters in regions previously declared Ebola free. Assays that have Emergency Use Authorization (EUA) and are routinely used to detect Ebola virus RNA in whole blood and plasma specimens at the Liberian Institute for Biomedical Research were tested for their suitability in detecting the presence of Ebola virus RNA in semen. Qiagen AVL extraction protocols, as well as the Ebola Zaire Target 1 and major groove binder quantitative reverse-transcription polymerase chain reaction assays, were demonstrably suitable for this purpose and should facilitate epidemiologic investigations, including those involving long-term survivors of Ebola.

  9. OPTIMAL DISTRIBUTED FUSION ALGORITHM WITH ONE-STEP OUT-OF-SEQUENCE ESTIMATES

    Institute of Scientific and Technical Information of China (English)

    Ge Quanbo; Wen Chenglin

    2008-01-01

    The transmission modes of multi-hop and broadcasting for Wireless Sensor Networks (WSN) often make random and unknown transmission delays appear, so multisensor data fusion based on delayed systems attracts intense attention from lots of researchers. The existing achievements for the delayed fusion all focus on Out-Of-Sequence Measurements (OOSM) problem which has many dis- advantages such as high communication cost, low computational efficiency, huge computational com- plexity and storage requirement, bad real-time performance and so on. In order to overcome these problems occurred in the OOSM fusion, the Out-Of-Sequence Estimates (OOSE) are considered to solve the delayed fusion for the first time. Different from OOSM which belongs to the centralized fusion, the OOSE scheme transmits local estimates from local sensors to the central processor and is thus the distributed fusion; thereby, the OOSE fusion can not only avoid the problems suffered in the OOSM fusion but also make the design of fusion algorithm highly simple and easy. Accordingly, a novel optimal linear recursive prediction weighted fusion method is proposed for one-step OOSE problem in this letter. As a tradeoff, its fusion accuracy is slightly lower than that of the OOSM method because the current OOSM fusion is a smooth estimate and OOSE gets a prediction estimate. But, the smooth result of the OOSE problem also has good fusion accuracy. Performance analysis and computer simulation show that the total performance of the proposed one-step OOSE fusion algorithm is better than the current one-step OOSM fusion in the practical tracking systems.

  10. A dissociative fluorescence enhancement technique for one-step time-resolved immunoassays

    Energy Technology Data Exchange (ETDEWEB)

    Blomberg, Kaj R.; Mukkala, Veli-Matti; Hakala, Harri H.O.; Maekinen, Pauliina H.; Suonpaeae, Mikko U.; Hemmilae, Ilkka A. [PerkinElmer Inc, Wallac Oy, P.O. Box 10, Turku (Finland)

    2011-02-15

    The limitation of current dissociative fluorescence enhancement techniques is that the lanthanide chelate structures used as molecular probes are not stable enough in one-step assays with high concentrations of complexones or metal ions in the reaction mixture since these substances interfere with lanthanide chelate conjugated to the detector molecule. Lanthanide chelates of diethylenetriaminepentaacetic acid (DTPA) are extremely stable, and we used EuDTPA derivatives conjugated to antibodies as tracers in one-step immunoassays containing high concentrations of complexones or metal ions. Enhancement solutions based on different {beta}-diketones were developed and tested for their fluorescence-enhancing capability in immunoassays with EuDTPA-labelled antibodies. Characteristics tested were fluorescence intensity, analytical sensitivity, kinetics of complex formation and signal stability. Formation of fluorescent complexes is fast (5 min) in the presented enhancement solution with EuDTPA probes withstanding strong complexones (ethylenediaminetetra acetate (EDTA) up to 100 mM) or metal ions (up to 200 {mu}M) in the reaction mixture, the signal is intensive, stable for 4 h and the analytical sensitivity with Eu is 40 fmol/L, Tb 130 fmol/L, Sm 2.1 pmol/L and Dy 8.5 pmol/L. With the improved fluorescence enhancement technique, EDTA and citrate plasma samples as well as samples containing relatively high concentrations of metal ions can be analysed using a one-step immunoassay format also at elevated temperatures. It facilitates four-plexing, is based on one chelate structure for detector molecule labelling and is suitable for immunoassays due to the wide dynamic range and the analytical sensitivity. (orig.)

  11. One step synthesis of 6-oxo-cholestan-3β,5α-diol

    Energy Technology Data Exchange (ETDEWEB)

    Voisin, Maud; Silvente-Poirot, Sandrine; Poirot, Marc, E-mail: marc.poirot@inserm.fr

    2014-04-11

    Highlights: • Cholesterol-5,6-epoxides are metabolized into cholestane-3β,5α,6β-triol (CT) in cancer cells. • 6-Oxo-cholestan-3β,5α-diol (OCDO) is a putative metabolite of CT. • The one step syntheses of CT and OCDO from cholesterol are reported. • The one step syntheses of labelled CT and OCDO are reported. - Abstract: Cholesterol metabolism has been recently linked to cancer, highlighting the importance of the characterization of new metabolic pathways in the sterol series. One of these pathways is centered on cholesterol-5,6-epoxides (5,6-ECs). 5,6-ECs can either generate dendrogenin A, a tumor suppressor present in healthy mammalian tissues, or the carcinogenic cholestane-3β,5α,6β-triol (CT) and its putative metabolite 6-oxo-cholestan-3β,5α-diol (OCDO) in tumor cells. We are currently investigating the identification of the enzyme involved in OCDO biosynthesis, which would be highly facilitated by the use of commercially unavailable [{sup 14}C]-cholestane-3β,5α,6β-triol and [{sup 14}C]-6-oxo-cholestan-3β,5α-diol. In the present study we report the one-step synthesis of [{sup 14}C]-cholestane-3β,5α,6β-triol and [{sup 14}C]-6-oxo-cholestan-3β,5α-diol by oxidation of [{sup 14}C]-cholesterol with iodide metaperiodate (HIO{sub 4})

  12. Influence of ageing on self-etch adhesives: one-step vs. two-step systems.

    Science.gov (United States)

    Marchesi, Giulio; Frassetto, Andrea; Visintini, Erika; Diolosà, Marina; Turco, Gianluca; Salgarello, Stefano; Di Lenarda, Roberto; Cadenaro, Milena; Breschi, Lorenzo

    2013-02-01

    The aim of this study was to evaluate microtensile bond strength (μTBS) to dentine, interfacial nanoleakage expression, and stability after ageing, of two-step vs. one-step self-etch adhesives. Human molars were cut to expose middle/deep dentine, assigned to groups (n = 15), and treated with the following bonding systems: (i) Optibond XTR (a two-step self-etch adhesive; Kerr), (ii) Clearfil SE Bond (a two-step self-etch adhesive; Kuraray), (iii) Adper Easy Bond (a one-step self-etch adhesive; 3M ESPE), and (iv) Bond Force (a one-step self-etch adhesive; Tokuyama). Specimens were processed for μTBS testing after 24 h, 6 months, or 1 yr of storage in artificial saliva at 37°C. Nanoleakage expression was examined in similarly processed additional specimens. At baseline the μTBS results ranked in the following order: Adper Easy Bond = Optibond XTR ≥Clearfil SE = Bond Force, and interfacial nanoleakage analysis showed Clearfil SE Bond = Adper Easy Bond = Optibond XTR> Bond Force. After 1 yr of storage, Optibond XTR, Clearfil SE Bond, and Adper Easy Bond showed higher μTBS and lower interfacial nanoleakage expression compared with Bond Force. In conclusion, immediate bond strength, nanoleakage expression, and stability over time were not related to the number of steps of the bonding systems, but to their chemical formulations. © 2012 Eur J Oral Sci.

  13. Long-term durability of one-step adhesive-composite systems to enamel and dentin.

    Science.gov (United States)

    Foxton, Richard M; Melo, Luciana; Stone, David G; Pilecki, Peter; Sherriff, Martin; Watson, Timothy F

    2008-01-01

    This study evaluated the long-term durability of three one-step adhesive-composite systems to ground enamel and dentin. Twenty-seven teeth were randomly divided into three groups of nine. The first group had its crowns sectioned to expose superficial dentin, which was then ground with 600 grit SiC paper. One of three one-step adhesives: a trial bonding agent, OBF-2; i Bond or Adper Prompt L-Pop was applied to the dentin of three teeth and built-up with the corresponding resin composite (Estelite sigma, Venus or Filtek Supreme). The second group of nine teeth had their enamel approximal surfaces ground with wet 600-grit SiC paper, then one of the three one-step adhesives was applied and built-up with resin composite. The bonded specimens were sliced into 0.7 mm-thick slabs. After 24 hours and one year of water storage at 37 degrees C, the slabs were sectioned into beams for the microtensile bond strength test. Failure modes were observed using optical and electron scanning microscopy. The third group of nine teeth had approximal wedge-shaped cavities prepared above the CEJ into dentin. Two-to-three grains of rhodamine B were added to each of the three adhesives prior to restoring the cavities with resin composite. After 24 hours storage, the teeth were sectioned and their interfaces examined with a laser scanning confocal microscope. The bond strengths of the three adhesive-composite systems to both enamel and dentin significantly lessened after one year of water storage, however, there was no significant difference between the materials.

  14. SVM with Quadratic Polynomial Kernel Function Based Nonlinear Model One-step-ahead Predictive Control

    Institute of Scientific and Technical Information of China (English)

    钟伟民; 何国龙; 皮道映; 孙优贤

    2005-01-01

    A support vector machine (SVM) with quadratic polynomial kernel function based nonlinear model one-step-ahead predictive controller is presented. The SVM based predictive model is established with black-box identification method. By solving a cubic equation in the feature space, an explicit predictive control law is obtained through the predictive control mechanism. The effect of controller is demonstrated on a recognized benchmark problem and on the control of continuous-stirred tank reactor (CSTR). Simulation results show that SVM with quadratic polynomial kernel function based predictive controller can be well applied to nonlinear systems, with good performance in following reference trajectory as well as in disturbance-rejection.

  15. One-step iterative reconstruction of conductivity inclusion via the concept of topological derivative

    CERN Document Server

    Park, Won-Kwang

    2013-01-01

    We consider an inverse problem of location identification of small conductivity inhomogeneity inside a conductor via boundary measurements which occurs in the EIT (Electrical Impedance Tomography). For this purpose, we derive topological derivative by applying the asymptotic formula for steady state voltage potentials in the existence of conductivity inclusion of small diameter. Using this derivative, we design only one-step iterative location search algorithm of small conductivity inhomogeneity completely embedded in the homogeneous domain by solving an adjoint problem. Numerical experiments presented for showing the feasibility of proposed algorithm.

  16. Fiber inline Michelson interferometer fabricated by one-step femtosecond laser micromachining for sensing applications

    Science.gov (United States)

    Yuan, Lei; Wu, Hongbin; Wang, Cong; Yu, Yingyu; Wang, Sumei; Xiao, Hai

    2013-12-01

    A fiber inline Michelson interferometer fiber optic sensor was presented for sensing applications, including high temperature performance and refractive index change. The sensor was fabricated using one-step femtosecond (fs) laser micromachining technique. A step structure at the tip of a single mode optical fiber was formed during the micromachining process. The device had a loss of 16 dB and an interference visibility exceeding 18 dB. The capability of this device for temperature sensing up to 1000 °C and refractive index sensing application in various concentrations of ethanol solution were all demonstrated.

  17. One-step synthesis of samarium-doped ceria and its CO catalysis

    Indian Academy of Sciences (India)

    Tian Deng; Chongrong Zhang; Yuyuan Xiao; Anran Xie; Yue Pang; Yanzhao Yang

    2015-09-01

    The samarium-doped ceria (SDC) nanospheres were prepared by the one-step hydrothermal method and characterized by transmission electron microscope, scanning electron microscope, powder X-ray diffraction, X-ray photoelectron spectroscopy, energy-dispersive spectrometer and Raman spectra. According to the results, samarium was doped into the lattice successfully. The as-prepared samples were dispersed well and the average diameter was 60 nm. It showed better catalytic performance than pure ceria and the most appropriate concentration of doping was found.

  18. Polymer Adsorption on Iron Oxide Nanoparticles for One-Step Amino-Functionalized Silica Encapsulation

    Directory of Open Access Journals (Sweden)

    Lionel Maurizi

    2015-01-01

    Full Text Available This paper presents an original method to obtain, in one step, core/shell nanoparticles grafted covalently with polymer and functionalized with amino groups. By combining polyvinyl alcohol (PVA and silica precursors, we were able to obtain silica-coated and amino-functionalized iron oxide nanoparticles (SPIONs cross-linked with PVA. We also showed that using silica and amino-silica precursors together significantly increased the amount of PVA covalently bonded to the SPION surface compared to using only silica precursors. This original and interesting method has high potential for the industrial development of biocompatible functionalized nanoparticles for targeting nanomedicine.

  19. One-step block method for solving Volterra integro-differential equations

    Science.gov (United States)

    Mohamed, Nurul Atikah binti; Majid, Zanariah Abdul

    2015-10-01

    One-step block method for solving linear Volterra integro-differential equations (VIDEs) is presented in this paper. In VIDEs, the unknown function appears in the form of derivative and under the integral sign. The popular methods for solving VIDEs are the method of quadrature or quadrature method combined with numerical method. The proposed block method will solve the ordinary differential equations (ODEs) part and Newton-Cotes quadrature rule is applied to calculate the integral part of VIDEs. Numerical problems are presented to illustrate the performance of the proposed method.

  20. One-step green synthesis of gold nanoparticles by mesophilic filamentous fungi

    Science.gov (United States)

    Vágó, Adél; Szakacs, George; Sáfrán, György; Horvath, Robert; Pécz, Béla; Lagzi, István

    2016-02-01

    We report a one-step and green method for the synthesis of gold nanoparticles by randomly selected 21 types of microscopic fungi. Depending on the exact type of microorganisms and experimental parameters (such as fermentation conditions) highly stable particles with various shapes and sizes were obtained from the cell-free extracts of fungi, as revealed by spectroscopic and electron microscopic measurements. The active compound in the fungal extracellular supernatants which is responsible for the bioreduction was found to be less than 3000 Da in molecular weight determined by molecular sieves.

  1. A ONE-STEP SURFACE MODIFICATION PROCEDURE TO PEG-GRAFTED NANO-TITANIA

    Institute of Scientific and Technical Information of China (English)

    Yong-liang Zhu; Bin Wang; Jian Yu; Zhao-xia Guo

    2007-01-01

    PEG (Polyethylene Glycol)-grafted nano-titania has been obtained in a one-step procedure using hexamethylene diisocyanate as the coupling agent and dibutyltin dilaurate as the catalyst in toluene at 80℃ and characterized qualitatively by FTIR and quantitatively by elemental analysis and thermogravimetric analysis. A comparison of nano-titania with two other commonly used inorganic nanoparticles, nano-silica and nano-alumina, is made, revealing that reactivity order is nanosilica > nano-alumina > nano-titania in view of PEG grafting. Possible mechanism of PEG grafting is also discussed.

  2. One-step Entanglement Generation Between Separated Nitrogen-vacancy Centers Embedded in Photonic Crystal Cavities

    Science.gov (United States)

    Santos, A. F.; Khanna, Faqir C.

    2016-09-01

    We propose a one-step scheme for creating entanglement between two distant nitrogen-vacancy (NV) centers, which are placed in separate single-mode nanocavities in a planar photonic crystal (PC). With a laser-driven, the decoherence from the excited states of the NV centers can be effectively suppressed by virtue of the Raman transition in the dispersive regime. With the assistant of a strong classical field, fast operation can be achieved. The experimental feasibility of the scheme is discussed based on currently available technology.

  3. Template-directed synthesis of flexible porphyrin nanocage and nanorings via one-step olefin metathesis.

    Science.gov (United States)

    Zhu, Bin; Chen, Huanxin; Lin, Wei; Ye, Yang; Wu, Jing; Li, Shijun

    2014-10-29

    We describe the fabrication of a suite of flexible porphyrin cages and nanorings from a simple tetraalkene-derived zinc porphyrin monomer via a highly efficient template-directed strategy. The zinc porphyrin monomers were preorganized together by coordination with N atoms of multidentate ligands. Subsequent one-step olefin metathesis furnished a bisporphyrin cage, a triporphyrin nanoring, and a hexaporphyrin nanoring. In the case of the hexaporphyrin nanoring, 24 terminal olefins from six porphyrin monomers reacted with each other to form 12 new double bonds, delivering the final product. The triporphyrin and hexaporphyrin nanorings were further employed as hosts to encapsulate C60 and C70.

  4. Optimized spray drying process for preparation of one-step calcium-alginate gel microspheres

    Energy Technology Data Exchange (ETDEWEB)

    Popeski-Dimovski, Riste [Department of physic, Faculty of Natural Sciences and Mathematics, “ss. Cyril and Methodius” University, Arhimedova 3, 1000 Skopje, R. Macedonia (Macedonia, The Former Yugoslav Republic of)

    2016-03-25

    Calcium-alginate micro particles have been used extensively in drug delivery systems. Therefore we establish a one-step method for preparation of internally gelated micro particles with spherical shape and narrow size distribution. We use four types of alginate with different G/M ratio and molar weight. The size of the particles is measured using light diffraction and scanning electron microscopy. Measurements showed that with this method, micro particles with size distribution around 4 micrometers can be prepared, and SEM imaging showed that those particles are spherical in shape.

  5. One-step generation of qutrit entanglement via adiabatic passage in cavity quantum electrodynamics

    Institute of Scientific and Technical Information of China (English)

    Ma Song-She; Chen Mei-Feng; Jiang Xia-Ping

    2011-01-01

    A scheme is proposed for generating a three-dimensional entangled state for two atoms trapped in a cavity by one step via adiabatic passage.In the scheme,the two atoms are always in ground states and the field mode of the cavity excited is negligible under a certain condition.Therefore,the scheme is very robust against decoherence.Furthermore,it needs neither the exact control of all parameters nor the accurate control of the interaction time.It is shown that qutrit entanglement can be generated with a high fidelity.

  6. Template-activated strategy toward one-step coating silica colloidal microspheres with sliver.

    Science.gov (United States)

    Wang, Ke; Zhang, Xiaoli; Niu, Chunyu; Wang, Yongqiang

    2014-01-22

    Template-activated strategy was developed to coat silica (SiO2) colloidal microspheres with silver in one step, based on one-pot hydrothermal treatment of silver nitrate, PVP (poly(vinyl pyrrolidone)), and SiO2 colloidal microspheres in ammonia solution. In our reaction system, the surface of SiO2 colloidal microspheres was continually activated with negative-charged SiO(-) groups in ammonia solution, which accumulated [Ag(NH3)2](+) or Ag(+) ions around the SiO2 colloidal microspheres through electrostatic attraction; thereafter these ions could be reduced into Ag nanoparticles in situ by the weak reducer PVP in the solution, and then acted as seeds for the subsequent complete silver coating with the reaction proceeding. Therefore, the traditional three steps for complete silver coating, including prior surface modification, seeding, and subsequent growing, were effectively integrated into one step. The experimental results exhibited that perfect SiO2/Ag core/shell composite microspheres were successfully synthesized through optimizing the reaction parameters like the solvent ingredient, reducer, and the reaction temperature. Additionally, these obtained uniform composite microspheres were further used as SERS substrate by using R6G and thiram as probe molecules, and showed excellent trace detection of these organic chemicals in solution.

  7. Operator Approach to the Master Equation for the One-Step Process

    Science.gov (United States)

    Hnatič, M.; Eferina, E. G.; Korolkova, A. V.; Kulyabov, D. S.; Sevastyanov, L. A.

    2016-02-01

    Background. Presentation of the probability as an intrinsic property of the nature leads researchers to switch from deterministic to stochastic description of the phenomena. The kinetics of the interaction has recently attracted attention because it often occurs in the physical, chemical, technical, biological, environmental, economic, and sociological systems. However, there are no general methods for the direct study of this equation. The expansion of the equation in a formal Taylor series (the so called Kramers-Moyal's expansion) is used in the procedure of stochastization of one-step processes. Purpose. However, this does not eliminate the need for the study of the master equation. Method. It is proposed to use quantum field perturbation theory for the statistical systems (the so-called Doi method). Results: This work is a methodological material that describes the principles of master equation solution based on quantum field perturbation theory methods. The characteristic property of the work is that it is intelligible for non-specialists in quantum field theory. Conclusions: We show the full equivalence of the operator and combinatorial methods of obtaining and study of the one-step process master equation.

  8. One-step synthesis of magnetic chitosan for controlled release of 5-hydroxytryptophan

    Energy Technology Data Exchange (ETDEWEB)

    Santos Menegucci, Jucély dos; Santos, Mac-Kedson Medeiros Salviano; Dias, Diego Juscelino Santos; Chaker, Juliano Alexandre; Sousa, Marcelo Henrique, E-mail: mhsqui@gmail.com

    2015-04-15

    In this work, nanoparticles of chitosan embedded with 25% (w/w) of iron oxide magnetic nanoparticles (magnetite/maghemite) with narrow size-distribution and with a loading efficiency of about 80% for 5-hydroxytryptophan (5-HTP), which is a chemical precursor in the biosynthesis of important neurotransmitters as serotonin, were synthesized with an initial mass ratio of 5-HTP/magnetic chitosan=1.2, using homogeneous precipitation by urea decomposition, in an efficient one-step procedure. Characterization of morphology, structure and surface were performed by XRD, TEM, FTIR, TGA, magnetization and zeta potential measurements, while drug loading and drug releasing were investigated using UV–vis spectroscopy. Kinetic drug release experiments under different pH conditions revealed a pH-sensitivecontrolled-release system, ruled by polymer swelling and/or particle dissolution. - Highlights: • One-step synthesis and incorporation of drug in magnetic chitosan. • Synthesis utilizes a cost-effective and environmentally friendly procedure. • Narrow size distribution of magnetic nanoparticles in the composite. • Composite is a basis for a magnetic pH triggered drug release system.

  9. One-step isolation of plasma membrane proteins using magnetic beads with immobilized concanavalin A1

    Science.gov (United States)

    Lee, Yu-Chen; Block, Gregory; Chen, Huiwen; Folch-Puy, Emma; Foronjy, Robert; Jalili, Roxana; Jendresen, Christian Bille; Kimura, Masashi; Kraft, Edward; Lindemose, Søren; Lu, Jin; McLain, Teri; Nutt, Leta; Ramon-Garcia, Santiago; Smith, Joseph; Spivak, Aaron; Wang, Michael L.; Zanic, Marija; Lin, Sue-Hwa

    2008-01-01

    We have developed a simple method for isolating and purifying plasma membrane proteins from various cell types. This one-step affinity-chromatography method uses the property of the lectin concanavalin A (ConA) and the technique of magnetic-bead separation to obtain highly purified plasma membrane proteins from crude membrane preparations or cell lines. ConA is immobilized onto magnetic beads by binding biotinylated ConA to streptavidin magnetic beads. When these ConA magnetic beads were used to enrich plasma membranes from a crude membrane preparation, this procedure resulted in 3.7-fold enrichment of plasma membrane marker 5′-nucleotidase activity with 70% recovery of the activity in the crude membrane fraction of rat liver. In agreement with the results of 5′-nucleotidase activity, immunoblotting with antibodies specific for a rat liver plasma membrane protein, CEACAM1, indicated that CEACAM1 was enriched about threefold relative to that of the original membranes. In similar experiments, this method produced 13-fold enrichment of 5′-nucleotidase activity with 45% recovery of the activity from a total cell lysate of PC-3 cells and 7.1-fold enrichment of 5′-nucleotidase activity with 33% recovery of the activity from a total cell lysate of HeLa cells. These results suggest that this one-step purification method can be used to isolate total plasma membrane proteins from tissue or cells for the identification of membrane biomarkers. PMID:18765283

  10. One-step ahead prediction of foF2 using time series forecasting techniques

    Directory of Open Access Journals (Sweden)

    A. Belehaki

    2005-11-01

    Full Text Available In this paper the problem of one-step ahead prediction of the critical frequency (foF2 of the middle-latitude ionosphere, using time series forecasting methods, is considered. The whole study is based on a sample of about 58000 observations of foF2 with 15-min time resolution, derived from the Athens digisonde ionograms taken from the Digisonde Portable Sounder (DPS4 located at Palaia Penteli (38° N, 23.5° E, for the period from October 2002 to May 2004. First, the embedding dimension of the dynamical system that generates the above sample is estimated using the false nearest neighbor method. This information is then utilized for the training of the predictors employed in this study, which are the linear predictor, the neural network predictor, the persistence predictor and the k-nearest neighbor predictor. The results obtained by the above predictors suggest that, as far as the mean square error is considered as performance criterion, the first two predictors are significantly better than the latter two predictors. In addition, the results obtained by the linear and the neural network predictors are not significantly different from each other. This may be taken as an indication that a linear model suffices for one step ahead prediction of foF2.

  11. A simple one-step chemistry model for partially premixed hydrocarbon combustion

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez-Tarrazo, Eduardo [Instituto Nacional de Tecnica Aeroespacial, Madrid (Spain); Sanchez, Antonio L. [Area de Mecanica de Fluidos, Universidad Carlos III de Madrid, Leganes 28911 (Spain); Linan, Amable [ETSI Aeronauticos, Pl. Cardenal Cisneros 3, Madrid 28040 (Spain); Williams, Forman A. [Department of Mechanical and Aerospace Engineering, University of California San Diego, La Jolla, CA 92093-0411 (United States)

    2006-10-15

    This work explores the applicability of one-step irreversible Arrhenius kinetics with unity reaction order to the numerical description of partially premixed hydrocarbon combustion. Computations of planar premixed flames are used in the selection of the three model parameters: the heat of reaction q, the activation temperature T{sub a}, and the preexponential factor B. It is seen that changes in q with equivalence ratio f need to be introduced in fuel-rich combustion to describe the effect of partial fuel oxidation on the amount of heat released, leading to a universal linear variation q(f) for f>1 for all hydrocarbons. The model also employs a variable activation temperature T{sub a}(f) to mimic changes in the underlying chemistry in rich and very lean flames. The resulting chemistry description is able to reproduce propagation velocities of diluted and undiluted flames accurately over the whole flammability limit. Furthermore, computations of methane-air counterflow diffusion flames are used to test the proposed chemistry under nonpremixed conditions. The model not only predicts the critical strain rate at extinction accurately but also gives near-extinction flames with oxygen leakage, thereby overcoming known predictive limitations of one-step Arrhenius kinetics. (author)

  12. Copper nano-clusters prepared by one-step electrodeposition and its application on nitrate sensing

    Directory of Open Access Journals (Sweden)

    Y. Li

    2015-04-01

    Full Text Available This paper describes the fabrication and characterization of copper nano-clusters prepared by a simple one-step electrodeposition process on platinum microelectrode, and the application for nitrate determination. The one-step electrodepostion process was performed by chronoamperometry scan in acidic copper sulphate electrolyte directly. The SEM and electrochemical examination showed that the morphologies and microstructures of deposited copper layers can be precisely controlled by using different deposition voltages. It was found that the copper layer is porous when the deposition voltage is higher than -500 mV, and this porous layer has a larger effective surface area compared with the corresponding smooth flat copper layer deposited under voltage less than -300 mV. Under the optimized deposition voltage, copper clusters constructed by uniform nanoparticles with an average diameter of about 100 nm can be obtained. The mechanism of electrodeposition process for this method was also speculated. The copper layers deposited under different voltages are used in a series of tests in order to evaluate their performance for nitrate sensing. The experimental results reveal that the microelectrode modified by fixed potential deposition under -700 mV had a higher sensitivity of 39.31 μA/mmolL−1 for nitrate detection within the concentration ranging from 0.1 mmolL−1 to 4.0 mmolL−1.

  13. One-Step Printable Perovskite Films Fabricated under Ambient Conditions for Efficient and Reproducible Solar Cells.

    Science.gov (United States)

    Jung, Yen-Sook; Hwang, Kyeongil; Heo, Youn-Jung; Kim, Jueng-Eun; Lee, Donmin; Lee, Cheol-Ho; Joh, Han-Ik; Yeo, Jun-Seok; Kim, Dong-Yu

    2017-08-23

    Despite the potential of roll-to-roll processing for the fabrication of perovskite films, the realization of highly efficient and reproducible perovskite solar cells (PeSCs) through continuous coating techniques and low-temperature processing is still challenging. Here, we demonstrate that efficient and reliable CH3NH3PbI3 (MAPbI3) films fabricated by a printing process can be achieved through synergetic effects of binary processing additives, N-cyclohexyl-2-pyrrolidone (CHP) and dimethyl sulfoxide (DMSO). Notably, these perovskite films are deposited from premixed perovskite solutions for facile one-step processing under a room-temperature and ambient atmosphere. The CHP molecules result in the uniform and homogeneous perovskite films even in the one-step slot-die system, which originate from the high boiling point and low vapor pressure of CHP. Meanwhile, the DMSO molecules facilitate the growth of perovskite grains by forming intermediate states with the perovskite precursor molecules. Consequently, fully printed PeSC based on the binary additive system exhibits a high PCE of 12.56% with a high reproducibility.

  14. One step 'dip' and 'use' Ag nanostructured thin films for ultrahigh sensitive SERS Detection.

    Science.gov (United States)

    Rajkumar, Kanakaraj; Jayram, Naidu Dhanpal; Mangalaraj, Devanesan; Rajendra Kumar, Ramasamy Thangavelu

    2016-11-01

    A simple one step galvanic displacement method which involves dipping of the silicon substrate in the AgNO3/HF solution and using it for SERS application without any further process is demonstrated. The size and shape of the Ag nanoparticles changes as the deposition time is increased. Initially the shape of the particles was nearly spherical and as it grows, becomes oblong and then coalesce to form a discontinuous film with vertically grown hierarchical Ag nanostructures. The sizes of the deposited particles were in the ranges from 30nm to a discontinuous film. It also demonstrated a highly sensitive chemical detection by surface-enhanced Raman scattering of rhodamine 6G dye, down to 10(-16)M concentration. Prepared samples were able to detect lower concentrations of Melamine. Discontinuous thin films with hierarchical Ag nanostructures were obtained for 5min Ag deposition. The formation of Hot spots between the discontinuous islands and also along the hierarchical structures is responsible for the high SERS enhancement. This simple one step, fast, non-lithographic and cost effective method can be applied for various label free detection of analytes of importance.

  15. One-step electrodeposition of graphene loaded nickel oxides nanoparticles for acetaminophen detection.

    Science.gov (United States)

    Liu, Gui-Ting; Chen, Hui-Fen; Lin, Guo-Ming; Ye, Ping-ping; Wang, Xiao-Ping; Jiao, Ying-Zhi; Guo, Xiao-Yu; Wen, Ying; Yang, Hai-Feng

    2014-06-15

    An electrochemical sensor of acetaminophen (AP) based on electrochemically reduced graphene (ERG) loaded nickel oxides (Ni2O3-NiO) nanoparticles coated onto glassy carbon electrode (ERG/Ni2O3-NiO/GCE) was prepared by a one-step electrodeposition process. The as-prepared electrode was characterized by scanning electron microscopy, X-ray photoelectron spectroscopy and Raman spectroscopy. The electrocatalytic properties of ERG/Ni2O3-NiO modified glassy carbon electrode toward the oxidation of acetaminophen were analyzed via cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The electrodes of Ni2O3-NiO/GCE, ERG/GCE, and Ni2O3-NiO deposited ERG/GCE were fabricated for the comparison and the catalytic mechanism understanding. The studies showed that the one-step prepared ERG/Ni2O3-NiO/GCE displayed the highest electro-catalytic activity, attributing to the synergetic effect derived from the unique composite structure and physical properties of nickel oxides nanoparticles and graphene. The low detection limit of 0.02 μM (S/N=3) with the wide linear detection range from 0.04 μM to 100 μM (R=0.998) was obtained. The resulting sensor was successfully used to detect acetaminophen in commercial pharmaceutical tablets and urine samples.

  16. General fabrication of ordered nanocone arrays by one-step selective plasma etching.

    Science.gov (United States)

    Wang, Qiang; Tian, Zhaoshuo; Li, Yunlong; Tian, Shibing; Li, Yunming; Ren, Shoutian; Gu, Changzhi; Li, Junjie

    2014-03-21

    One-step selective direct current (DC) plasma etching technology is employed to fabricate large-area well-aligned nanocone arrays on various functional materials including semiconductor, insulator and metal. The cones have nanoscale apexes (∼2 nm) with high aspect ratios, which were achieved by a selective plasma etching process using only CH4 and H2 in a bias-assisted hot filament chemical vapor deposition (HFCVD) system without any masked process. The CH(3)(+) ions play a major role to etch the roughened surface into a conical structure under the auxiliary of H(+) ions. Randomly formed nano-carbon may act as an original mask on the smooth surface to initiate the following selective ions sputtering. Physical impinging of energetic ions onto the concave regions is predominant in comparison with the etching of convex parts on the surface, which is identified as the key mechanism for the formation of conical nanostructures. This one-step maskless plasma etching technology enables the universal formation of uniform nanocone structures on versatile substrates for many promising applications.

  17. Transparent, superhydrophobic surfaces from one-step spin coating of hydrophobic nanoparticles.

    Science.gov (United States)

    Xu, Lebo; Karunakaran, Raghuraman G; Guo, Jia; Yang, Shu

    2012-02-01

    We study the nonwettability and transparency from the assembly of fluorosilane modified silica nanoparticles (F-SiO(2) NPs) via one-step spin-coating and dip-coating without any surface postpassivation steps. When spin-coating the hydrophobic NPs (100 nm in diameter) at a concentration ≥ 0.8 wt % in a fluorinated solvent, the surface exhibited superhydrophobicity with an advancing water contact angle greater than 150° and a water droplet (5 μL) roll-off angle less than 5°. In comparison, superhydrophobicity was not achieved by dip-coating the same hydrophobic NPs. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) images revealed that NPs formed a nearly close-packed assembly in the superhydrophobic films, which effectively minimized the exposure of the underlying substrate while offering sufficiently trapped air pockets. In the dip-coated films, however, the surface coverage was rather random and incomplete. Therefore, the underlying substrate was exposed and water was able to impregnate between the NPs, leading to smaller water contact angle and larger water contact angle hysteresis. The spin-coated superhydrophobic film was also highly transparent with greater than 95% transmittance in the visible region. Further, we demonstrated that the one-step coating strategy could be extended to different polymeric substrates, including poly(methyl methacrylate) and polyester fabrics, to achieve superhydrophobicity.

  18. A one-step miniprep for the isolation of plasmid DNA and lambda phage particles.

    Directory of Open Access Journals (Sweden)

    George Lezin

    Full Text Available Plasmid DNA minipreps are fundamental techniques in molecular biology. Current plasmid DNA minipreps use alkali and the anionic detergent SDS in a three-solution format. In addition, alkali minipreps usually require additional column-based purification steps and cannot isolate other extra-chromosomal elements, such as bacteriophages. Non-ionic detergents (NIDs have been used occasionally as components of multiple-solution plasmid DNA minipreps, but a one-step approach has not been developed. Here, we have established a one-tube, one-solution NID plasmid DNA miniprep, and we show that this approach also isolates bacteriophage lambda particles. NID minipreps are more time-efficient than alkali minipreps, and NID plasmid DNA performs better than alkali DNA in many downstream applications. In fact, NID crude lysate DNA is sufficiently pure to be used in digestion and sequencing reactions. Microscopic analysis showed that the NID procedure fragments E. coli cells into small protoplast-like components, which may, at least in part, explain the effectiveness of this approach. This work demonstrates that one-step NID minipreps are a robust method to generate high quality plasmid DNA, and NID approaches can also isolate bacteriophage lambda particles, outperforming current standard alkali-based minipreps.

  19. One-step fabrication of near superhydrophobic aluminum surface by nanosecond laser ablation

    Science.gov (United States)

    Jagdheesh, R.; García-Ballesteros, J. J.; Ocaña, J. L.

    2016-06-01

    Inspired by the micro and nano structures of biological surface such as lotus leaf, rice leaves, etc. a functional near superhydrophobic surface of pure aluminum has been fabricated using one-step nanosecond laser processing. Thin aluminum sheets are micro-patterned with ultraviolet laser pulses to create near superhydrophobic surface in one-step direct laser writing technique. The impact of number of pulses/microhole with respect to the geometry and static contact angle measurements has been investigated. The microstructure shows the formation of blind microholes along with the micro-wall by laser processing, which improves the composite interface between the three phases such as water, air and solid, thus enhance the wetting property of the surface. The geometrical changes are supported by the chemical changes induced on the surface for improving the degree of hydrophobicity. Laser processed microholes exhibited near superhydrophobic surface with SCA measurement of 148 ± 3°. The static contact angle values are very consistent for repeated measurement at same area and across the laser patterned surface.

  20. Shape-control and characterization of iron nanocrystals prepared via a one-step solvothermal method

    Energy Technology Data Exchange (ETDEWEB)

    Fan, Junmei, E-mail: jmfan@home.ipe.ac.cn [State Key Laboratory of Multi-phase Complex System, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190 (China); Cao, Yuebin [Key Laboratory for Advanced Technology in Environmental Protection of Jiangsu Province, Yancheng Institute of Technology, Yancheng 224051 (China); Yuan, Fangli [State Key Laboratory of Multi-phase Complex System, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190 (China)

    2015-12-01

    In the present work, iron nanocubes, nanorods and nanowires are successfully synthesized by one-step reduction approach in a solvothermal environment. It is analyzed that the iron nanocubes, nanorods and nanowires belong to the pure body-centered cubic structure of α-Fe. Effects of additive sort and the amount of Anionic Gemini surfactant 12-4-12 on the morphology evolution of Fe are discussed based on SEM and TEM images. The possible formation mechanisms of iron nanocubes, nanorods and nanowires are proposed. The Anionic Gemini surfactant 12-4-12 regarded as template plays an important role in the formation of iron nanocrystals. The microwave electromagnetic (EM) (80 wt% Fe) and calculated microwave-absorbing (0.5 mm thickness) properties in 2–18 GHz of the iron nanocubes, nanorods and nanowires are studied systematically. Results show that iron nanowires are superior to nanorods and nanocubes, which indicates the potential to be a super-thin microwave absorber. - Highlights: • Iron nanocubes, nanorods and nanowires were synthesized by one-step reduction. • The Anionic Gemini played a vital role in the formation of iron nanocrystals. • The microwave-absorbing properties in 2–18 GHz of iron nanowires were tiptop. • Iron nanowires have the potential to be a super-thin microwave absorber.

  1. One-step synthesis of nitrogen-iron coordinated carbon nanotube catalysts for oxygen reduction reaction

    Science.gov (United States)

    Choi, Woongchul; Yang, Gang; Kim, Suk Lae; Liu, Peng; Sue, Hung-Jue; Yu, Choongho

    2016-05-01

    Prohibitively expensive precious metal catalysts for oxygen reduction reaction (ORR) have been one of the major hurdles in a wide use of electrochemical cells. Recent significant efforts to develop precious metal free catalysts have resulted in excellent catalytic activities. However, complicated and time-consuming synthesis processes have negated the cost benefit. Moreover, detailed analysis about catalytically active sites and the role of each element in these high-performance catalysts containing nanomaterials for large surface areas are often lacking. Here we report a facile one-step synthesis method of nitrogen-iron coordinated carbon nanotube (CNT) catalysts without precious metals. Our catalysts show excellent long-term stability and onset ORR potential comparable to those of other precious metal free catalysts, and the maximum limiting current density from our catalysts is larger than that of the Pt-based catalysts. We carry out a series of synthesis and characterization experiments with/without iron and nitrogen in CNT, and identify that the coordination of nitrogen and iron in CNT plays a key role in achieving the excellent catalytic performances. We anticipate our one-step process could be used for mass production of precious metal free electrocatalysts for a wide range of electrochemical cells including fuel cells and metal-air batteries.

  2. One-step preparation of antimicrobial silver nanoparticles in polymer matrix

    Energy Technology Data Exchange (ETDEWEB)

    Lyutakov, O., E-mail: lyutakoo@vscht.cz; Kalachyova, Y. [Institute of Chemical Technology, Department of Solid State Engineering (Czech Republic); Solovyev, A. [Institute of Chemical Process Fundamentals of the ASCR (Czech Republic); Vytykacova, S. [Institute of Chemical Technology, Department of Glass and Ceramics (Czech Republic); Svanda, J.; Siegel, J. [Institute of Chemical Technology, Department of Solid State Engineering (Czech Republic); Ulbrich, P. [Institute of Chemical Technology, Department of Biochemistry and Microbiology (Czech Republic); Svorcik, V. [Institute of Chemical Technology, Department of Solid State Engineering (Czech Republic)

    2015-03-15

    Simple one-step procedure for in situ preparation of silver nanoparticles (AgNPs) in the polymer thin films is described. Nanoparticles (NPs) were prepared by reaction of N-methyl pyrrolidone with silver salt in semi-dry polymer film and characterized by transmission electron microscopy, XPS, and UV–Vis spectroscopy techniques. Direct synthesis of NPs in polymer has several advantages; even though it avoids time-consuming NPs mixing with polymer matrix, uniform silver distribution in polymethylmethacrylate (PMMA) films is achieved without necessity of additional stabilization. The influence of the silver concentration, reaction temperature and time on reaction conversion rate, and the size and size-distribution of the AgNPs was investigated. Polymer films doped with AgNPs were tested for their antibacterial activity on Gram-negative bacteria. Antimicrobial properties of AgNPs/PMMA films were found to be depended on NPs concentration, their size and distribution. Proposed one-step synthesis of functional polymer containing AgNPs is environmentally friendly, experimentally simple and extremely quick. It opens up new possibilities in development of antimicrobial coatings with medical and sanitation applications.

  3. One-step disposal of Cr (Ⅵ)-bearing wastewater by natural pyrrhotite

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Cr(Ⅵ)-bearing wastewater can be treated by natural pyrrhotite which is used for reductant to reduce Cr(Ⅵ) and precipitant to precipitate Cr(Ⅲ) simultaneously. The disposal products can be divided into three parts in the beakers,namely supernatant in the upper part,the yellowish colloidal precipitates in the middle part and the pyrrhotite in the lower part. The content of total Cr=Cr(Ⅵ)+Cr(Ⅲ) in the supernatant liquid is 0.06 mg/L,which is lower than 1.5 mg/L of the discharge standard of China and near to 0.05 mg/L of the standard of potable water. This one-step disposal composing of both reduction and precipitation which is traditionally divided into two independent steps called reducing technology and precipitating technology respectively. The new method is of obvious economic advantage and favourable to decreasing surplus mud derived from adding Ca(OH)2 to precipitate Cr(Ⅲ) traditionally so as to avoid recontamination. In fact,sodium sulfite (Na2SO3) used in disposal of Cr(Ⅵ) was traditionally produced from natural mineral of pyrrhotite (FeS). One molecule of FeS is 4 times more than that of Na2SO3 from a view point of rational use of mineral resources. Therefore the prospective of application of the one-step disposal of Cr(Ⅵ) method is full of promise.

  4. Influence of application methods of one-step self-etching adhesives on microtensile bond strength

    Directory of Open Access Journals (Sweden)

    Chul-Kyu Choi,

    2011-05-01

    Full Text Available Objectives The purpose of this study was to evaluate the effect of various application methods of one-step self-etch adhesives to microtensile resin-dentin bond strength. Materials and Methods Thirty-six extracted human molars were used. The teeth were assigned randomly to twelve groups (n = 15, according to the three different adhesive systems (Clearfil Tri-S Bond, Adper Prompt L-Pop, G-Bond and application methods. The adhesive systems were applied on the dentin as follows: 1 The single coating, 2 The double coating, 3 Manual agitation, 4 Ultrasonic agitation. Following the adhesive application, light-cure composite resin was constructed. The restored teeth were stored in distilled water at room temperature for 24 hours, and prepared 15 specimens per groups. Then microtensile bond strength was measured and the failure mode was examined. Results Manual agitation and ultrasonic agitation of adhesive significantly increased the microtensile bond strength than single coating and double coating did. Double coating of adhesive significantly increased the microtensile bond strength than single coating did and there was no significant difference between the manual agitation and ultrasonic agitation group. There was significant difference in microtensile bonding strength among all adhesives and Clearfil Tri-S Bond showed the highest bond strength. Conclusions In one-step self-etching adhesives, there was significant difference according to application methods and type of adhesives. No matter of the material, the manual or ultrasonic agitation of the adhesive showed significantly higher microtensile bond strength.

  5. A Pipeline with Multiplex Reverse Transcription Polymerase Chain Reaction and Microarray for Screening of Chromosomal Translocations in Leukemia

    Directory of Open Access Journals (Sweden)

    Fei-Fei Xiong

    2013-01-01

    Full Text Available Chromosome rearrangements and fusion genes present major portion of leukemogenesis and contribute to leukemic subtypes. It is practical and helpful to detect the fusion genes in clinic diagnosis of leukemia. Present application of reverse transcription polymerase chain reaction (RT-PCR method to detect the fusion gene transcripts is effective, but time- and labor-consuming. To set up a simple and rapid system, we established a method that combined multiplex RT-PCR and microarray. We selected 15 clinically most frequently observed chromosomal rearrangements generating more than 50 fusion gene variants. Chimeric reverse primers and chimeric PCR primers containing both gene-specific and universal sequences were applied in the procedure of multiplex RT-PCR, and then the PCR products hybridized with a designed microarray. With this approach, among 200 clinic samples, 63 samples were detected to have gene rearrangements. All the detected fusion genes positive and negative were validated with RT-PCR and Sanger sequencing. Our data suggested that the RT-PCR-microarray pipeline could screen 15 partner gene pairs simultaneously at the same accuracy of the fusion gene detection with regular RT-PCR. The pipeline showed effectiveness in multiple fusion genes screening in clinic samples.

  6. Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification

    Directory of Open Access Journals (Sweden)

    Joojin Jeong

    2015-09-01

    Full Text Available The primary step for efficient control of viral diseases is the development of simple, rapid, and sensitive virus detection. Reverse transcription loop-mediated isothermal amplification (RT-LAMP has been used to detect viral RNA molecules because of its simplicity and high sensitivity for a number of viruses. RT-LAMP for the detection of Potato virus X (PVX was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR to demonstrate its advantages over RT-PCR. RT-LAMP reactions were conducted with or without a set of loop primers since one out of six primers showed PVX specificity. Based on real-time monitoring, RT-LAMP detected PVX around 30 min, compared to 120 min for RT-PCR. By adding a fluorescent reagent during the reaction, the extra step of visualization by gel electrophoresis was not necessary. RT-LAMP was conducted using simple inexpensive instruments and a regular incubator to evaluate whether RNA could be amplified at a constant temperature instead of using an expensive thermal cycler. This study shows the potential of RT-LAMP for the diagnosis of viral diseases and PVX epidemiology because of its simplicity and rapidness compared to RT-PCR.

  7. Rapid and specific detection of tdh, trh1, and trh2 mRNA of Vibrio parahaemolyticus by transcription-reverse transcription concerted reaction with an automated system.

    Science.gov (United States)

    Nakaguchi, Yoshitsugu; Ishizuka, Tetsuya; Ohnaka, Satoru; Hayashi, Toshinori; Yasukawa, Kiyoshi; Ishiguro, Takahiko; Nishibuchi, Mitsuaki

    2004-09-01

    Vibrio parahaemolyticus strains carrying the thermostable direct hemolysin (TDH) tdh gene, the TDH-related hemolysin (trh) gene, or both genes are considered virulent strains. We previously demonstrated that the transcription-reverse transcription concerted (TRC) method could be used to quantify the amount of mRNA transcribed from the tdh gene by using an automated detection system. In this study, we devised two TRC-based assays to quantify the mRNAs transcribed from the trh1 and trh2 genes, the two representative trh genes. The TRC-based detection assays for the tdh, trh1, and trh2 transcripts could specifically and quantitatively detect 10(3) to 10(7) copies of the corresponding calibrator RNAs. We examined by the three TRC assays the total RNA preparations extracted from 103 strains of Vibrio parahaemolyticus carrying the tdh, trh1, or trh2 gene in various combinations. The tdh, trh1, and trh2 mRNAs in the total RNA preparations were specifically quantified, and the time needed for detection ranged from 9 to 19 min, from 14 to 18 min, and from 9 to 12 min, respectively. The results showed that this automated TRC assays could detect the tdh, trh1, and trh2 mRNAs specifically, quantitatively, and rapidly. The relative levels of TDH determined by the immunological method and that of tdh mRNA determined by the TRC assays for most tdh-positive strains correlated. Interestingly, the levels of TDH produced from the strains carrying both tdh and trh genes were lower than those carrying only the tdh gene, whereas the levels of mRNA did not significantly differ between the two groups.

  8. [Reversed effect of valproic acid on transcription inhibition of AML1-ETO fusion protein of kasumi-1 leukemic cell line].

    Science.gov (United States)

    Zhao, Lei; Zhu, Cui-Min; Zhang, Zhi-Hua; Tian, Wen-Liang; Hao, Chang-Lai

    2009-04-01

    This study was aimed to investigate the mechanism of histone deacetylase (HDAC) inhibitor, valproic acid (VPA), reversing transcription inhibition of AML1-ETO fusion protein in Kasumi-1 cell line. The mRNA expressions of AML1-ETO, AML1 and cyclin D2 were detected by semi-quantitation RT-PCR after treating kasumi-1 cells with VPA at different doses/and different time points. The results indicated that the mRNA expression of AML1-ETO showed no obvious change, when kasumi-1 cells were treated with VPA. Compared with control group, the expression level of AML1 mRNA significantly increased in a dose-dependent manner. Compared with control group, the expression level of cyclin D2 mRNA significantly decreased when kasumi-1 cells had been treated with 3 mmol/L VPA as well as kasumi-1 cells were treated with different concentrations of VPA for 3 days. In conclusion, VPA could remove transcription inhibition of AML1-ETO fusion protein, increase transcription of AML1 and down-regulate mRNA expression of AML1 target gene cyclin D2 through HDAC inhibiting activity.

  9. Hypothesis: Artifacts, Including Spurious Chimeric RNAs with a Short Homologous Sequence, Caused by Consecutive Reverse Transcriptions and Endogenous Random Primers.

    Science.gov (United States)

    Peng, Zhiyu; Yuan, Chengfu; Zellmer, Lucas; Liu, Siqi; Xu, Ningzhi; Liao, D Joshua

    2015-01-01

    Recent RNA-sequencing technology and associated bioinformatics have led to identification of tens of thousands of putative human chimeric RNAs, i.e. RNAs containing sequences from two different genes, most of which are derived from neighboring genes on the same chromosome. In this essay, we redefine "two neighboring genes" as those producing individual transcripts, and point out two known mechanisms for chimeric RNA formation, i.e. transcription from a fusion gene or trans-splicing of two RNAs. By our definition, most putative RNA chimeras derived from canonically-defined neighboring genes may either be technical artifacts or be cis-splicing products of 5'- or 3'-extended RNA of either partner that is redefined herein as an unannotated gene, whereas trans-splicing events are rare in human cells. Therefore, most authentic chimeric RNAs result from fusion genes, about 1,000 of which have been identified hitherto. We propose a hypothesis of "consecutive reverse transcriptions (RTs)", i.e. another RT reaction following the previous one, for how most spurious chimeric RNAs, especially those containing a short homologous sequence, may be generated during RT, especially in RNA-sequencing wherein RNAs are fragmented. We also point out that RNA samples contain numerous RNA and DNA shreds that can serve as endogenous random primers for RT and ensuing polymerase chain reactions (PCR), creating artifacts in RT-PCR.

  10. One-Step Electrochemical Polymerization of Polyaniline Flexible Counter Electrode Doped by Graphene

    Directory of Open Access Journals (Sweden)

    Qi Qin

    2016-01-01

    Full Text Available To improve the photoelectric property of polyaniline (PANI counter electrode using for flexible dye-sensitized solar cell (DSSC, graphene (GN was doped in PANI films covered on flexible conducting substrate by one-step electrochemical method, and then GN/PANI composites are characterized by scanning electron microscope (SEM, fourier transform infrared spectroscopy (FTIR, four probe instrument, and so on. The results show that PANI particles can be electrodeposited on the surface of GN sheets as the potential rising to 2.0 V. This formed unique PANI-GN-PANI lamellar structure owing to the strong interaction of conjugated π electron between GN and PANI results in the superior conductivity and catalytic performance of GN/PANI electrode. The maximum conversion efficiency of dye-sensitized solar cell with this counter electrode reaches 4.31%, which is much higher than that of GN-free PANI counter electrode.

  11. Properties and Microstructure of One-Step Cold Rolled Steel Strip for Shadow Mask

    Institute of Scientific and Technical Information of China (English)

    LIANG Xuan; LI Jun; PENG Ying-hong

    2008-01-01

    The effect of recrystallization annealing temperature on the properties and microstructure of one-step cold rolled steel strip for shadow mask was studied.The results showed that there was no yield point elongation when the tensile tests were performed on the samples for annealing temperatures ranging from 750℃ to 810 ℃.Moreover,increasing annealing temperature resulted in large grains,which was beneficial to the formability and magnetic property of steel strips.On the other hand,when the sample was annealed at 840 ℃,its microstructure showed ununifortuity with 0.04% yield point elongation,which was not good for the function of the shadow mask.Therefore,the proper recrystallization annealing temperature was about 810 ℃ for the present steel strip for shadow mask.

  12. One-step synthesis of magnetic chitosan for controlled release of 5-hydroxytryptophan

    Science.gov (United States)

    Santos Menegucci, Jucély dos; Santos, Mac-Kedson Medeiros Salviano; Dias, Diego Juscelino Santos; Chaker, Juliano Alexandre; Sousa, Marcelo Henrique

    2015-04-01

    In this work, nanoparticles of chitosan embedded with 25% (w/w) of iron oxide magnetic nanoparticles (magnetite/maghemite) with narrow size-distribution and with a loading efficiency of about 80% for 5-hydroxytryptophan (5-HTP), which is a chemical precursor in the biosynthesis of important neurotransmitters as serotonin, were synthesized with an initial mass ratio of 5-HTP/magnetic chitosan=1.2, using homogeneous precipitation by urea decomposition, in an efficient one-step procedure. Characterization of morphology, structure and surface were performed by XRD, TEM, FTIR, TGA, magnetization and zeta potential measurements, while drug loading and drug releasing were investigated using UV-vis spectroscopy. Kinetic drug release experiments under different pH conditions revealed a pH-sensitivecontrolled-release system, ruled by polymer swelling and/or particle dissolution.

  13. ABS polymer electroless plating through a one-step poly(acrylic acid) covalent grafting.

    Science.gov (United States)

    Garcia, Alexandre; Berthelot, Thomas; Viel, Pascal; Mesnage, Alice; Jégou, Pascale; Nekelson, Fabien; Roussel, Sébastien; Palacin, Serge

    2010-04-01

    A new, efficient, palladium- and chromium-free process for the electroless plating of acrylonitrile-butadiene-styrene (ABS) polymers has been developed. The process is based on the ion-exchange properties of poly(acrylic acid) (PAA) chemically grafted onto ABS via a simple and one-step method that prevents using classical surface conditioning. Hence, ABS electroless plating can be obtained in three steps, namely: (i) the grafting of PAA onto ABS, (ii) the copper Cu(0) seeding of the ABS surface, and (iii) the nickel or copper metallization using commercial-like electroless plating bath. IR, XPS, and SEM were used to characterize each step of the process, and the Cu loading was quantified by atomic absorption spectroscopy. This process successfully compares with the commercial one based on chromic acid etching and palladium-based seed layer, because the final metallic layer showed excellent adhesion with the ABS substrate.

  14. One step paired electrochemical synthesis of iron and iron oxide nanoparticles

    Directory of Open Access Journals (Sweden)

    Ordoukhanian Juliet

    2016-09-01

    Full Text Available In this study, a new one step paired electrochemical method is developed for simultaneous synthesis of iron and iron oxide nanoparticles. iron and iron oxide are prepared as cathodic and anodic products from iron (ii sulfate aqueous solution in a membrane divided electrolytic cell by the pulsed current electrosynthesis. Because of organic solvent-free and electrochemical nature of the synthesis, the process could be considered as green and environmentally friendly. The reduction of energy consumption and low cost are the other significant advantages of this new method that would have a great application potential in the chemical industry. The nanostructure of prepared samples was characterized by Fourier transform infrared spectroscopy (FT-IR, X-ray diffraction (XRD, scanning electron microscopy (SEM and transmission electron microscopy (TEM. The magnetic properties were studied by vibrating sample magnetometer (VsM.

  15. One-step Double-layer Thermal Evaporation Method for Organic Light Emitting Diodes

    Science.gov (United States)

    Kee, Y. Y.; Yong, T. K.; Ong, D. S.; Tou, T. Y.

    2011-03-01

    A new one-step double-layer thermal evaporation method was used to fabricate organic light emitting diodes (OLEDs) with device structure of: ITO (anode)/N,N_-diphenyl-N,N_-bis(3-methylphenyl)-1,1_-diphenyl-4,4_-diamine (TPD) /tris-(8-hydroxyquinoline)aluminum(3) (Alq3)/Al (cathode). These OLEDs were fabricated in cleanroom on the ITO-coated glass with a sheet resistivity of 20Ω/sq and an optical transmittance of 90%. The I-V and brightness characteristic showed that the new method could produce better performance achieving lower turn-on voltage (-2V), higher peak current efficiency (+29%) and higher brightness (+36%).

  16. Superlinear/Quadratic One-step Smoothing Newton Method for P0-NCP

    Institute of Scientific and Technical Information of China (English)

    Li Ping ZHANG; Ji Ye HAN; Zheng Hai HUANG

    2005-01-01

    We propose a one-step smoothing Newton method for solving the non-linear complementarity problem with P0-function (P0-NCP) based on the smoothing symmetric perturbed Fisher function (for short, denoted as the SSPF-function). The proposed algorithm has to solve only one linear system of equations and performs only one line search per iteration. Without requiring any strict complementarity assumption at the P0-NCP solution, we show that the proposed algorithm converges globally and superlinearly under mild conditions. Furthermore, the algorithm has local quadratic convergence under suitable conditions. The main feature of our global convergence results is that we do not assume a priori the existence of an accumulation point. Compared to the previous literatures, our algorithm has stronger convergence results under weaker conditions.

  17. One-step preparation of nitrogen-doped graphene nanosheets for high-performance supercapacitors

    Science.gov (United States)

    Gao, Biaofeng; Zhou, Haitao; Yang, Jianhong

    2017-07-01

    Graphene nanosheets (GNs) have been successfully synthesized by one-step carbonization and simultaneous chemical activation of polyaniline (PANI) nanofibers, with large surface area (1759.5 m2 g-1), high pore volume (1.16 cm3 g-1) and a mesopore rich structure (volume in micropore/total ratio, 2.16%). The vital (NDs) become entrapped within a carbon lattice (to form GNs). GNs could be used as an outstanding electrode material for supercapacitors in EMIMBF4 with high cycling stability (more than 73% retention of the initial capacitance after 10,000 cycles). The symmetric supercapacitor has an excellent energy density of 170.2 Wh kg-1 (25 °C) at 0.5 A g-1, with an operating cell voltage of 4.0 V. Meanwhile, oriented activation with polymers of anisotropic geometry might be regarded as a novel process to prepare GNs.

  18. One-Step Facile Synthesis of a Simple Hole Transport Material for Efficient Perovskite Solar Cells

    KAUST Repository

    Chen, Hu

    2016-04-04

    A hole transporting material was designed for use in perovskite solar cells, with a facile one-step synthesis from inexpensive, com-mercially available reagents. The molecule comprises a central fluorinated phenyl core with pendant aryl amines, namely, 3,6-difluoro-N1,N1,N2,N2,N4,N4,N5,N5-octakis(4-methoxyphenyl)benzene-1,2,4,5-tetraamine (DFTAB). A power conversion efficiency of up to 10.4% was achieved in a mesoporous perovskite device architecture. The merits of a simple and potentially low cost syn-thetic route as well as promising performance in perovskite devices, encourages further development of this materials class as new low-cost hole transporting materials for the scale up of perovskite solar cells.

  19. One-Step Fabrication of Microchannels with Integrated Three Dimensional Features by Hot Intrusion Embossing

    Directory of Open Access Journals (Sweden)

    Mike Debono

    2016-11-01

    Full Text Available We build on the concept of hot intrusion embossing to develop a one-step fabrication method for thermoplastic microfluidic channels containing integrated three-dimensional features. This was accomplished with simple, rapid-to-fabricate imprint templates containing microcavities that locally control the intrusion of heated thermoplastic based on their cross-sectional geometries. The use of circular, rectangular and triangular cavity geometries was demonstrated for the purposes of forming posts, multi-focal length microlense arrays, walls, steps, tapered features and three-dimensional serpentine microchannels. Process variables, such as temperature and pressure, controlled feature dimensions without affecting the overall microchannel geometry. The approach was demonstrated for polycarbonate, cycloolefin copolymer and polystyrene, but in principle is applicable to any thermoplastic. The approach is a step forward towards rapid fabrication of complex, robust, microfluidic platforms with integrated multi-functional elements.

  20. Cellulose acetate from oil palm empty fruit bunch via a one step heterogeneous acetylation.

    Science.gov (United States)

    Wan Daud, Wan Rosli; Djuned, Fauzi Muhammad

    2015-11-05

    Acetone soluble oil palm empty fruit bunch cellulose acetate (OPEFB-CA) of DS 2.52 has been successfully synthesized in a one-step heterogeneous acetylation of OPEFB cellulose without necessitating the hydrolysis stage. This has only been made possible by the mathematical modeling of the acetylation process by manipulating the variables of reaction time and acetic anhydride/cellulose ratio (RR). The obtained model was verified by experimental data with an error of less than 2.5%. NMR analysis showed that the distribution of the acetyl moiety among the three OH groups of cellulose indicates a preference at the C6 position, followed by C3 and C2. XRD revealed that OPEFB-CA is highly amorphous with a degree of crystallinity estimated to be ca. 6.41% as determined from DSC. The OPEFB-CA films exhibited good mechanical properties being their tensile strength and Young's modulus higher than those of the commercial CA.

  1. One-step preparation of zeolite silicalite-1 microspheres with adjustable macroporosity

    KAUST Repository

    Hua, Jia

    2009-06-23

    A facile one-step method was developed for the preparation of zeolite silicalite-1 (or ZSM-5) microspheres without the need of pre-synthesizing zeolite nanocrystals. In this method, Pluronic triblock copolymer F127 was added into a homogeneous silicalite-1 synthesis solution. During the hydrothermal synthesis, zeolite nanocrystals (∼ 100 nm) were formed and spontaneously assembled into uniform micrometer-sized (3-5 μm) spheres due to the presence of F127. The obtained microspheres possess significant textual porosity (up to 0.24 cm 3/g), which can be adjusted by simply adding different amounts of styrene in the synthesis. The zeolite microspheres are useful for enzyme immobilization, and the enzyme loading is proportional to the textual porosity. © 2009 American Chemical Society.

  2. One-Step Synthesis and Photoluminescence Eva-luation of Cadmium-containing Quantum Dots

    Institute of Scientific and Technical Information of China (English)

    Nisha Shukla; Michael M Nigra; Abigail D Ondeck

    2012-01-01

    We have developed a simple one-step process for synthesis of ternary quantum dots (ZnCdSe, MgCdSe) with photoluminescence wavelengths ranging from the red to the blue region of the visible spectrum. The primary aim of this work was to develop a synthesis for the preparation of Cd-containing quantum dots using a Cd precursor with lower toxicity than those used in common syntheses. This synthesis makes use of Cd(acac)2 which is significantly less toxic than precursors such as CdO and CdCl2 . We have studied the effect of solvent boiling point, precursors and reaction time on the photoluminescence properties of the ternary quantum dots. Ternary quantum dots synthesized from Cd(acac)2 in low boiling point solvents have photoluminescence wavelengths in the blue region, while those synthesized in high boiling point solvents have photoluminescence wavelengths in the red region.

  3. One-Step Method for Preparation of Magnetic Nanoparticles Coated with Chitosan

    Directory of Open Access Journals (Sweden)

    Karla M. Gregorio-Jauregui

    2012-01-01

    Full Text Available Preparation of magnetic nanoparticles coated with chitosan in one step by the coprecipitation method in the presence of different chitosan concentrations is reported here. Obtaining of magnetic superparamagnetic nanoparticles was confirmed by X-ray diffraction and magnetic measurements. Scanning transmission electron microscopy allowed to identify spheroidal nanoparticles with around 10-11 nm in average diameter. Characterization of the products by Fourier transform infrared spectroscopy demonstrated that composite chitosan-magnetic nanoparticles were obtained. Chitosan content in obtained nanocomposites was estimated by thermogravimetric analysis. The nanocomposites were tested in Pb2+ removal from a PbCl2 aqueous solution, showing a removal efficacy up to 53.6%. This work provides a simple method for chitosan-coated nanoparticles obtaining, which could be useful for heavy metal ions removal from water.

  4. One step effective removal of Congo Red in chitosan nanoparticles by encapsulation

    Science.gov (United States)

    Alver, Erol; Bulut, Mehmet; Metin, Ayşegül Ülkü; Çiftçi, Hakan

    2017-01-01

    Chitosan nanoparticles (CNPs) were prepared with ionotropic gelation between chitosan and tripolyphosphate for the removal of Congo Red. The production of chitosan nanoparticles and the dye removal process was carried out in one-step. The removal efficiency of Congo Red by encapsulation within chitosan from the aqueous solution and its storage stability are examined at different pH values. The influence of some parameters such as the initial dye concentration, pH value of the dye solution, electrolyte concentration, tripolyphosphate concentration, mixing time and speed on the encapsulation is examined. Congo Red removal efficiency and encapsulation capacity of chitosan nanoparticles were determined as above 98% and 5107 mg Congo Red/g chitosan, respectively.

  5. One-step synthesis of highly reduced graphene hydrogels for high power supercapacitor applications

    Science.gov (United States)

    Banda, Harish; Aradilla, David; Benayad, Anass; Chenavier, Yves; Daffos, Barbara; Dubois, Lionel; Duclairoir, Florence

    2017-08-01

    Graphene hydrogels with high electrical conductivity were prepared by a one-step process using hydrazine hydrate as gel assembly agent (GH-HD). Conventional two-step process of gel formation and further reduction to prepare highly conducting gels was replaced by a single step involving equivalent amount of hydrazine. Optimized graphene oxide concentration was established to facilitate such monolith formation. Extensive characterization and control studies enabled understanding of the material properties and gel formation mechanism. The synthesized gel shows a high electrical conductivity of 1141 S/m. The supercapacitor based on GH-HD delivers a high specific capacitance of 190 F/g at a current density of 0.5 A/g and 123 F/g at very high current density of 100 A/g. Furthermore, excellent power capability and cyclic stability were also observed. 3D macroporous morphology of GH-HD makes it ideal for high rate supercapacitor applications.

  6. Centrifuge modeling of one-step outflow tests for unsaturated parameter estimations

    Directory of Open Access Journals (Sweden)

    H. Nakajima

    2006-05-01

    Full Text Available Centrifuge modeling of one-step outflow tests were carried out using a 2-m radius geotechnical centrifuge, and the cumulative outflow and transient pore pressure were measured during the tests at multiple gravity levels. Based on the scaling law of centrifuge modeling, the measurements generally showed reasonable agreement with prototype data calculated from forward simulations with input parameters determined from standard laboratory tests. The parameter optimizations were examined for three different combinations of input data sets using the test measurements. Within the gravity level examined in this study up to 40 g, the optimized unsaturated parameters compared well when accurate pore pressure measurements were included along with cumulative outflow as input data. The centrifuge modeling technique with its capability to implement variety of instrumentations under well controlled initial and boundary conditions, shortens testing time and can provide significant information for the parameter estimation procedure.

  7. Efficient hydrogen evolution in transition metal dichalcogenides via a simple one-step hydrazine reaction

    Science.gov (United States)

    Cummins, Dustin R.; Martinez, Ulises; Sherehiy, Andriy; Kappera, Rajesh; Martinez-Garcia, Alejandro; Schulze, Roland K.; Jasinski, Jacek; Zhang, Jing; Gupta, Ram K.; Lou, Jun; Chhowalla, Manish; Sumanasekera, Gamini; Mohite, Aditya D.; Sunkara, Mahendra K.; Gupta, Gautam

    2016-01-01

    Hydrogen evolution reaction is catalysed efficiently with precious metals, such as platinum; however, transition metal dichalcogenides have recently emerged as a promising class of materials for electrocatalysis, but these materials still have low activity and durability when compared with precious metals. Here we report a simple one-step scalable approach, where MoOx/MoS2 core-shell nanowires and molybdenum disulfide sheets are exposed to dilute aqueous hydrazine at room temperature, which results in marked improvement in electrocatalytic performance. The nanowires exhibit ∼100 mV improvement in overpotential following exposure to dilute hydrazine, while also showing a 10-fold increase in current density and a significant change in Tafel slope. In situ electrical, gate-dependent measurements and spectroscopic investigations reveal that hydrazine acts as an electron dopant in molybdenum disulfide, increasing its conductivity, while also reducing the MoOx core in the core-shell nanowires, which leads to improved electrocatalytic performance. PMID:27282871

  8. Chitosan-Coated Magnetic Nanoparticles with Low Chitosan Content Prepared in One-Step

    Directory of Open Access Journals (Sweden)

    Yolanda Osuna

    2012-01-01

    Full Text Available Chitosan-coated magnetic nanoparticles (CMNP were obtained at 50°C in a one-step method comprising coprecipitation in the presence of low chitosan content. CMNP showed high magnetization and superparamagnetism. They were composed of a core of 9.5 nm in average diameter and a very thin chitosan layer in accordance with electron microscopy measurements. The results from Fourier transform infrared spectrometry demonstrated that CMNP were obtained and those from thermogravimetric analysis allowed to determine that they were composed of 95 wt% of magnetic nanoparticles and 5 wt% of chitosan. 67% efficacy in the Pb+2 removal test indicated that only 60% of amino groups on CMNP surface bound to Pb, probably due to some degree of nanoparticle flocculation during the redispersion. The very low weight ratio chitosan to magnetic nanoparticles obtained in this study, 0.053, and the high yield of the precipitation reactions (≈97% are noticeable.

  9. Ionic liquid-stabilized non-spherical gold nanofluids synthesized using a one-step method.

    Science.gov (United States)

    Zhang, Hao; Cui, Hua; Yao, Shiwei; Zhang, Kelong; Tao, Haikun; Meng, Haibo

    2012-10-23

    Ionic liquid (IL)-stabilized non-spherical gold nanofluids have been synthesized by a one-step method in aqueous solution. The whole reaction proceeded in room temperature. In the presence of amino-functionalized ionic liquids, gold nanofluids with long-wave surface plasmon resonance (SPR) absorption (>600 nm) could be obtained by adopting tannic acid as the reductant. The specific SPR absorption was related to the non-spherical gold nanoparticles including gold triangle, decahedra, and icosahedra nanocrystals. All the nanocrystals were observed by transmission electron microscopy. It was deduced that the formation of non-spherical gold nanofluids was related to the hydroxyls in tannic acid while IL acted as the synthesis template.

  10. One-step GPS for the estimation of temperature-dependent thermal conductivity

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Chein-Shan [Department of Mechanical and Mechatronic Engineering, Taiwan Ocean University, Keelung 202-24 (China)

    2006-08-15

    In this paper we are concerned with the estimation of temperature-dependent thermal conductivity of a one-dimensional inverse heat conduction problem. First, we construct a one-step group-preserving scheme (GPS) for the semi-discretization of quasilinear heat conduction equation, and then derive a quasilinear algebraic equation to determine the unknown thermal conductivity under a given initial temperature and a measured temperature perturbed by noise at time T. The new method does not require any prior information on the functional form of thermal conductivity. Several examples are examined to show that the new approach has high accuracy and efficiency, and the number of iterations spent in solving the quasilinear algebraic equation is smaller than five even in a large temperature range. (author)

  11. One-Step Fabrication of Microchannels with Integrated Three Dimensional Features by Hot Intrusion Embossing

    Science.gov (United States)

    Debono, Mike; Voicu, Dan; Pousti, Mohammad; Safdar, Muhammad; Young, Robert; Kumacheva, Eugenia; Greener, Jesse

    2016-01-01

    We build on the concept of hot intrusion embossing to develop a one-step fabrication method for thermoplastic microfluidic channels containing integrated three-dimensional features. This was accomplished with simple, rapid-to-fabricate imprint templates containing microcavities that locally control the intrusion of heated thermoplastic based on their cross-sectional geometries. The use of circular, rectangular and triangular cavity geometries was demonstrated for the purposes of forming posts, multi-focal length microlense arrays, walls, steps, tapered features and three-dimensional serpentine microchannels. Process variables, such as temperature and pressure, controlled feature dimensions without affecting the overall microchannel geometry. The approach was demonstrated for polycarbonate, cycloolefin copolymer and polystyrene, but in principle is applicable to any thermoplastic. The approach is a step forward towards rapid fabrication of complex, robust, microfluidic platforms with integrated multi-functional elements. PMID:27916849

  12. New one step functionalization of polycrystalline diamond films using amine derivatives

    Energy Technology Data Exchange (ETDEWEB)

    Agnes, Charles; Ruffinatto, Sebastien; Delbarre, Emma; Roget, Andre; Arnault, Jean-Charles; Omnes, Franck; Mailley, Pascal, E-mail: pascal.mailley@cea.fr, E-mail: franck.omnes@grenoble.cnrs.fr

    2010-11-15

    Diamond received tremendous interest for analytical sciences due to its intrinsic properties. However, the analytical perception of chemical environment requires surface functionalization that brings selectivity to the detection event. Thereby, many works focused on diamond modification using chemical or biochemical entities. We proposed here, a new and straightforward methodology for diamond (bio)functionalization. This method involves the chemical reaction between (bio)chemical entities presenting a primary amine moiety, used as grafting site, and hydrogenated diamond surface. This reaction allows in one step to modify diamond surface whatever its doping level and its crystalline quality. The effectiveness of this new method is exposed here through the grafting of one redox species, ferrocene, and of one biochemical, biotin. The impacts of both functionalization duration and pH are investigated and the robustness of the formed bond is demonstrated owing to biotin-avidin coupling.

  13. Centrifuge modeling of one-step outflow tests for unsaturated parameter estimations

    Directory of Open Access Journals (Sweden)

    H. Nakajima

    2006-01-01

    Full Text Available Centrifuge modeling of one-step outflow tests were carried out using a 2-m radius geotechnical centrifuge, and the cumulative outflow and transient pore water pressure were measured during the tests at multiple gravity levels. Based on the scaling laws of centrifuge modeling, the measurements generally showed reasonable agreement with prototype data calculated from forward simulations with input parameters determined from standard laboratory tests. The parameter optimizations were examined for three different combinations of input data sets using the test measurements. Within the gravity level examined in this study up to 40g, the optimized unsaturated parameters compared well when accurate pore water pressure measurements were included along with cumulative outflow as input data. With its capability to implement variety of instrumentations under well controlled initial and boundary conditions and to shorten testing time, the centrifuge modeling technique is attractive as an alternative experimental method that provides more freedom to set inverse problem conditions for the parameter estimation.

  14. One-Step Synthesis of Monodisperse In-Doped ZnO Nanocrystals

    Directory of Open Access Journals (Sweden)

    Wang QingLing

    2010-01-01

    Full Text Available Abstract A method for the synthesis of high quality indium-doped zinc oxide (In-doped ZnO nanocrystals was developed using a one-step ester elimination reaction based on alcoholysis of metal carboxylate salts. The resulting nearly monodisperse nanocrystals are well-crystallized with typically crystal structure identical to that of wurtzite type of ZnO. Structural, optical, and elemental analyses on the products indicate the incorporation of indium into the host ZnO lattices. The individual nanocrystals with cubic structures were observed in the 5% In–ZnO reaction, due to the relatively high reactivity of indium precursors. Our study would provide further insights for the growth of doped oxide nanocrystals, and deepen the understanding of doping process in colloidal nanocrystal syntheses.

  15. Fluorinated alkyne-derived monolayers on oxide-free silicon nanowires via one-step hydrosilylation

    Science.gov (United States)

    Nguyen Minh, Quyen; Pujari, Sidharam P.; Wang, Bin; Wang, Zhanhua; Haick, Hossam; Zuilhof, Han; van Rijn, Cees J. M.

    2016-11-01

    Passivation of oxide-free silicon nanowires (Si NWs) by the formation of high-quality fluorinated 1-hexadecyne-derived monolayers with varying fluorine content has been investigated. Alkyl chain monolayers (C16H30-xFx) with a varying number of fluorine substituents (x = 0, 1, 3, 9, 17) were attached onto hydrogen-terminated silicon (Sisbnd H) surfaces with an effective one-step hydrosilylation. This surface chemistry gives well-defined monolayers on nanowires that have a cylindrical core-shell structure, as characterized by X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FT-IR) and static contact angle (SCA) analysis. The monolayers were stable under acidic and basic conditions, as well as under extreme conditions (such as UV exposure), and provide excellent surface passivation, which opens up applications in the fields of field effect transistors, optoelectronics and especially for disease diagnosis.

  16. Corrosion Properties of Polydopamine Coatings Formed in One-Step Immersion Process on Magnesium.

    Science.gov (United States)

    Singer, Ferdinand; Schlesak, Magdalena; Mebert, Caroline; Höhn, Sarah; Virtanen, Sannakaisa

    2015-12-09

    Polydopamine layers were polymerized directly from Tris(hydroxymethyl)aminomethane-buffered solution in a one-step immersion process onto magnesium surface. Scanning electron microscopy showed successful formation of a ∼1 μm thick layer. ASTM D3359-09 "Tape test" revealed excellent adhesion of the layer. X-ray induced photoelectron spectroscopy and Fourier transform infrared spectroscopy verified the presence of polydopamine on the surface. Corrosion measurements were performed in 0.1 M NaCl solution investigating the influence of coating parameters: dopamine concentration, immersion time, solution pH, and immersion angle. Tafel analysis revealed strong improvement of corrosion behavior compared to bare magnesium. Polydopamine layers prepared with optimized coating procedure showed promising corrosion properties in Dulbecco's modified Eagle medium. In summary, polydopamine coatings offer a simple treatment for magnesium to improve the corrosion behavior and could further act as intermediate layer for further surface functionalization.

  17. One-step electro-spinning/netting technique for controllably preparing polyurethane nano-fiber/net.

    Science.gov (United States)

    Hu, Juanping; Wang, Xianfeng; Ding, Bin; Lin, Jinyou; Yu, Jianyong; Sun, Gang

    2011-11-01

    Electro-spinning/netting (ESN) as a cutting-edge technique evokes much interest because of its ability in the one-step preparation of versatile nano-fiber/net (NFN) membranes. Here, a controllable fabrication of polyurethane (PU) NFN membranes with attractive structures, consisting of common electrospun nanofibers and two-dimensional (2D) soap bubble-like structured nano-nets via an ESN process is reported. The unique nanoscaled NFN architecture can be finely controlled by regulating the solution properties and several ESN process parameters. The versatile PU nano-nets comprising interlinked nanowires with ultrathin diameters (5-40 nm) mean that the NFN structured membranes possess several excellent characteristics, such as an extremely large specific surface area, high porosity and large stacking density, which would be particularly useful for applications in ultrafiltration, special protective clothing, ultrasensitive sensors, catalyst support and so on.

  18. Shape-control and characterization of iron nanocrystals prepared via a one-step solvothermal method

    Science.gov (United States)

    Fan, Junmei; Cao, Yuebin; Yuan, Fangli

    2015-12-01

    In the present work, iron nanocubes, nanorods and nanowires are successfully synthesized by one-step reduction approach in a solvothermal environment. It is analyzed that the iron nanocubes, nanorods and nanowires belong to the pure body-centered cubic structure of α-Fe. Effects of additive sort and the amount of Anionic Gemini surfactant 12-4-12 on the morphology evolution of Fe are discussed based on SEM and TEM images. The possible formation mechanisms of iron nanocubes, nanorods and nanowires are proposed. The Anionic Gemini surfactant 12-4-12 regarded as template plays an important role in the formation of iron nanocrystals. The microwave electromagnetic (EM) (80 wt% Fe) and calculated microwave-absorbing (0.5 mm thickness) properties in 2-18 GHz of the iron nanocubes, nanorods and nanowires are studied systematically. Results show that iron nanowires are superior to nanorods and nanocubes, which indicates the potential to be a super-thin microwave absorber.

  19. One-step Patterning of Sub-wavelength Plasmonic Gratings in Metal-Polymer Composites

    CERN Document Server

    Chaudhary, Raghvendra P; Jaiswal, Arun; Hawal, Suyog R; Saxena, Sumit; Shukla, Shobha

    2016-01-01

    2D and 3D micro/nano fabrication based on two-photon polymerization (TPP) has emerged as a strong contender for additive manufacturing for wide variety of applications. In this manuscript we report one step patterning of structurally stable, subwavelength 2D and 3D gold nanostructures using femto-second laser by incorporating single photon photoinitiator only in pure and metal precursor doped polymers. The metal polymer composite nanostructures are written directly by in-situ reduction of gold precursor within the photoresist using femto-second laser irradiation. The photo-initiator triggers the reduction of gold precursor and induces simultaneous polymerization of the photoresist based on two-photon absorption phenomenon. Diffraction gratings with varied loading of gold precursors in photoresist have been fabricated and characterized by measuring their diffraction efficiencies in the infrared region. Minimum line width of 390 nm has been achieved for 5 wt% gold loaded polymers. Systematic studies of the effe...

  20. Preparation of Thermo-Responsive Poly(ionic liquids-Based Nanogels via One-Step Cross-Linking Copolymerization

    Directory of Open Access Journals (Sweden)

    Jing Zhang

    2015-09-01

    Full Text Available In this study, thermo-responsive polymeric nanogels were facilely prepared via one-step cross-linking copolymerization of ethylene glycol dimethacrylate/divinylbenzene and ionic liquid (IL-based monomers, 1,n-dialkyl-3,3′-bis-1-vinyl imidazolium bromides ([CnVIm]Br; n = 6, 8, 12 in selective solvents. The results revealed that stable and blue opalescent biimidazolium (BIm-based nanogel solutions could be obtained without any precipitation when the copolymerizations were conducted in methanol. Most importantly, these novel nanogels were thermo-response, and could reversibly transform to precipitation in methanol with temperature changes. Turbidity analysis and dynamic light scatting (DLS measurement illustrated that PIL-based nanogel solutions presented the phase transform with upper critical solution temperature (UCST in the range of 5–25 °C. The nanogels were characterized using Fourier transform infrared (FTIR, thermogravimetric analyses (TGA, and scanning electron microscopy (SEM. In addition, BIm-based nanogels could also be used as highly active catalysts in the cycloaddition reaction of CO2 and epoxides. As a result, our attributes build a robust platform suitable for the preparation of polymeric nanomaterials, as well as CO2 conversion.

  1. Nanotexturing of Conjugated Polymers via One-Step Maskless Oxygen Plasma Etching for Enhanced Tunable Wettability.

    Science.gov (United States)

    Jiang, Youhua; Xu, Jian; Lee, Junghoon; Du, Ke; Yang, Eui-Hyeok; Moon, Myoung-Woon; Choi, Chang-Hwan

    2017-07-11

    A one-step maskless oxygen plasma etching process is investigated to nanopattern conjugated polymer dodecylbenzenesulfonate doped polypyrrole (PPy(DBS)) and to examine the effects of nanostructures on the inherent tunable wettability of the surface and the droplet mobility. Etching characteristics such as the geometry and dimensions of the nanostructures are systematically examined for the etching power and duration. The mechanism of self-formation of vertically aligned dense-array pillared nanostructures in the one-step maskless oxygen plasma etching process is also investigated. Results show that lateral dimensions such as the periodicity and diameter of the pillared nanostructures are insensitive to the etching power and duration, whereas the length and aspect ratio of the nanostructures increase with them. X-ray photoelectron spectroscopy analysis and thermal treatment of the polymer reveal that the codeposition of impurities on the surface resulting from the holding substrate is the primary reason for the self-formation of nanostructures during the oxygen plasma etching, whereas the local crystallinity subject to thermal treatment has a minor effect on the lateral dimensions. Retaining the tunable wettability (oleophobicity) for organic droplets during the electrochemical redox (i.e., reduction and oxidization) process, the nanotextured PPy(DBS) surface shows significant enhancement of droplet mobility compared to that of the flat PPy(DBS) surface with no nanotexture by making the surface superoleophobic (i.e., in a Cassie-Baxter wetting state). Such enhancement of the tunable oleophobicity and droplet mobility of the conjugated polymer will be of great significance in many applications such as microfluidics, lab-on-a-chip devices, and water/oil treatment.

  2. One-step green synthesis of bimetallic Fe/Pd nanoparticles used to degrade Orange II

    Energy Technology Data Exchange (ETDEWEB)

    Luo, Fang; Yang, Die; Chen, Zuliang, E-mail: zuliang.chen@newcastle.edu.au; Megharaj, Mallavarapu; Naidu, Ravendra

    2016-02-13

    Highlights: • Green synthesis of bimetallic Fe/Pd NPs was firstly reported using the one-step method. • 98.0% of Orange II was removed by Fe/Pd NPs, but only 16.0% by Fe NPs. • Fe/Pd NPs with a diameter ranging from 10 to 100 nm were observed. • Removing Orange II using Fe/Pd NPs involved both adsorption and catalytic degradation. - Abstract: To reduce cost and enhance reactivity, bimetallic Fe/Pd nanoparticles (NPs) were firstly synthesized using grape leaf aqueous extract to remove Orange II. Green synthesized bimetallic Fe/Pd NPs (98.0%) demonstrated a far higher ability to remove Orange II in 12 h compared to Fe NPs (16.0%). Meanwhile, all precursors, e.g., grape leaf extract, Fe{sup 2+} and Pd{sup 2+}, had no obvious effect on removing Orange II since less than 2.0% was removed. Kinetics study revealed that the removal rate fitted well to the pseudo-first-order reduction and pseudo-second-order adsorption model, meaning that removing Orange II via Fe/Pd NPs involved both adsorption and catalytic reduction. The remarkable stability of Fe/Pd NPs showed the potential application for removing azo dyes. Furthermore, Scanning Electron Microscopy (SEM) and Fourier Transform Infrared Spectroscopy (FTIR) confirmed the changes in Fe/Pd NPs before and after reaction with Orange II. High Performance Liquid Chromatography–Mass Spectrum (HPLC–MS) identified the degraded products in the removal of Orange II, and finally a removal mechanism was proposed. This one-step strategy using grape leaf aqueous extract to synthesize Fe/Pd NPs is simple, cost-effective and environmentally benign, making possible the large-scale production of Fe/Pd NPs for field remediation.

  3. Facile one-step fabrication of magnetite particles under mild hydrothermal conditions

    Energy Technology Data Exchange (ETDEWEB)

    Keerthana, D. Shanthini; Namratha, K. [Centre for Materials Science and Technology, Vijnana Bhavan, University of Mysore, Manasagangothri, Mysore 6 (India); Byrappa, K., E-mail: kbyrappa@gmail.com [Centre for Materials Science and Technology, Vijnana Bhavan, University of Mysore, Manasagangothri, Mysore 6 (India); Yathirajan, H.S. [Centre for Materials Science and Technology, Vijnana Bhavan, University of Mysore, Manasagangothri, Mysore 6 (India); DOS in Chemistry, University of Mysore, Manasagangothri, Mysore 6 (India)

    2015-03-15

    Hydrophilic magnetite particles for biological applications were synthesized by hydrothermal method in the presence of D-Glucose as both reducing and capping agent in a facile, one-step, low energy and environmentally friendly route. The role of D-Glucose as a reducing agent in the formation of magnetite particles under mild hydrothermal conditions has been investigated. The absence of D-Glucose results in the formation of hematite. The magnetite particles synthesized were characterized using powder X-ray Diffraction (XRD), Fourier Transform Infrared (FTIR) Spectroscopy, High Resolution Scanning Electron Microscopy (HR-SEM), Dynamic Light Scattering (DLS) and Vibrating Sample Magnetometery (VSM). The influence of the quantity of D-Glucose used and the reaction duration on the formation of magnetite were studied. DLS and HR-SEM results show that the size of the particles was in nano- to micron range. The antioxidant potency of the particles was confirmed using DPPH assay, where 2,2- Diphenyl-1-picrylhydrazyl was used as a source of free radicals. Hence the magnetite particles obtained could be considered for the use in various biological applications. - Highlights: • Magnetite and hematite particles were synthesized using hydrothermal route. • The reactants and experimental parameters were optimized to get single phase magnetite particles. • The role of D-Glucose as a reducing and capping agent is discussed. • The magnetite particles showed very good free radical scavenging ability. • The authors report a simple, facile, one-step method using hydrated ferric salt as a single iron precursor.

  4. Recombinant Passenger Proteins Can Be Conveniently Purified by One-Step Affinity Chromatography.

    Directory of Open Access Journals (Sweden)

    Hua-zhen Wang

    Full Text Available Fusion tag is one of the best available tools to date for enhancement of the solubility or improvement of the expression level of recombinant proteins in Escherichia coli. Typically, two consecutive affinity purification steps are often necessitated for the purification of passenger proteins. As a fusion tag, acyl carrier protein (ACP could greatly increase the soluble expression level of Glucokinase (GlcK, α-Amylase (Amy and GFP. When fusion protein ACP-G2-GlcK-Histag and ACP-G2-Amy-Histag, in which a protease TEV recognition site was inserted between the fusion tag and passenger protein, were coexpressed with protease TEV respectively in E. coli, the efficient intracellular processing of fusion proteins was achieved. The resulting passenger protein GlcK-Histag and Amy-Histag accumulated predominantly in a soluble form, and could be conveniently purified by one-step Ni-chelating chromatography. However, the fusion protein ACP-GFP-Histag was processed incompletely by the protease TEV coexpressed in vivo, and a large portion of the resulting target protein GFP-Histag aggregated in insoluble form, indicating that the intracellular processing may affect the solubility of cleaved passenger protein. In this context, the soluble fusion protein ACP-GFP-Histag, contained in the supernatant of E. coli cell lysate, was directly subjected to cleavage in vitro by mixing it with the clarified cell lysate of E. coli overexpressing protease TEV. Consequently, the resulting target protein GFP-Histag could accumulate predominantly in a soluble form, and be purified conveniently by one-step Ni-chelating chromatography. The approaches presented here greatly simplify the purification process of passenger proteins, and eliminate the use of large amounts of pure site-specific proteases.

  5. Recombinant Passenger Proteins Can Be Conveniently Purified by One-Step Affinity Chromatography.

    Science.gov (United States)

    Wang, Hua-zhen; Chu, Zhi-zhan; Chen, Chang-chao; Cao, Ao-cheng; Tong, Xin; Ouyang, Can-bin; Yuan, Qi-hang; Wang, Mi-nan; Wu, Zhong-kun; Wang, Hai-hong; Wang, Sheng-bin

    2015-01-01

    Fusion tag is one of the best available tools to date for enhancement of the solubility or improvement of the expression level of recombinant proteins in Escherichia coli. Typically, two consecutive affinity purification steps are often necessitated for the purification of passenger proteins. As a fusion tag, acyl carrier protein (ACP) could greatly increase the soluble expression level of Glucokinase (GlcK), α-Amylase (Amy) and GFP. When fusion protein ACP-G2-GlcK-Histag and ACP-G2-Amy-Histag, in which a protease TEV recognition site was inserted between the fusion tag and passenger protein, were coexpressed with protease TEV respectively in E. coli, the efficient intracellular processing of fusion proteins was achieved. The resulting passenger protein GlcK-Histag and Amy-Histag accumulated predominantly in a soluble form, and could be conveniently purified by one-step Ni-chelating chromatography. However, the fusion protein ACP-GFP-Histag was processed incompletely by the protease TEV coexpressed in vivo, and a large portion of the resulting target protein GFP-Histag aggregated in insoluble form, indicating that the intracellular processing may affect the solubility of cleaved passenger protein. In this context, the soluble fusion protein ACP-GFP-Histag, contained in the supernatant of E. coli cell lysate, was directly subjected to cleavage in vitro by mixing it with the clarified cell lysate of E. coli overexpressing protease TEV. Consequently, the resulting target protein GFP-Histag could accumulate predominantly in a soluble form, and be purified conveniently by one-step Ni-chelating chromatography. The approaches presented here greatly simplify the purification process of passenger proteins, and eliminate the use of large amounts of pure site-specific proteases.

  6. One-step chromatographic purification of Helicobacter pylori neutrophil-activating protein expressed in Bacillus subtilis.

    Directory of Open Access Journals (Sweden)

    Kuo-Shun Shih

    Full Text Available Helicobacter pylori neutrophil-activating protein (HP-NAP, a major virulence factor of Helicobacter pylori (H. pylori, is capable of activating human neutrophils to produce reactive oxygen species (ROS and secrete inammatory mediators. HP-NAP is a vaccine candidate, a possible drug target, and a potential in vitro diagnostic marker for H. pylori infection. HP-NAP has also been shown to be a novel therapeutic agent for the treatment of allergic asthma and bladder cancer. Hence, an efficient way to obtain pure HP-NAP needs to be developed. In this study, one-step anion-exchange chromatography in negative mode was applied to purify the recombinant HP-NAP expressed in Bacillus subtilis (B. subtilis. This purification technique was based on the binding of host cell proteins and/or impurities other than HP-NAP to DEAE Sephadex resins. At pH 8.0, almost no other proteins except HP-NAP passed through the DEAE Sephadex column. More than 60% of the total HP-NAP with purity higher than 91% was recovered in the flow-through fraction from this single-step DEAE Sephadex chromatography. The purified recombinant HP-NAP was further demonstrated to be a multimeric protein with a secondary structure of α-helix and capable of activating human neutrophils to stimulate ROS production. Thus, this one-step negative chromatography using DEAE Sephadex resin can efficiently yield functional HP-NAP from B. subtilis in its native form with high purity. HP-NAP purified by this method could be further utilized for the development of new drugs, vaccines, and diagnostics for H. pylori infection.

  7. One-step surgical placement of Brånemark implants: a prospective multicenter clinical study.

    Science.gov (United States)

    Becker, W; Becker, B E; Israelson, H; Lucchini, J P; Handelsman, M; Ammons, W; Rosenberg, E; Rose, L; Tucker, L M; Lekholm, U

    1997-01-01

    This prospective longitudinal multicenter study evaluated the clinical outcomes after placement and restoration of one-step Brånemark implants into the maxillae and mandibles of completely and partially edentulous patients. Six surgical treatment centers participated in this study, in which 135 implants were placed into 63 adult patients. All implants were stable after placement. The majority of implants were placed into type B bone with minimal jaw resorption and type 2 bone quality. After implant placement, standard transmucosal healing abutments were firmly placed. The average amount of time between implant placement and prosthetic abutment connection was 170 days in the maxillae and 147 days in the mandibles. To evaluate crestal bone changes caused by implant placement, a periodontal probe was used to measure midbuccally from the top of the implant cylinder to the alveolar crest; in 29 patients, 54 midbuccal bone crest sites were remeasured following prosthetic abutment connection. Crestal bone changes in mandibles and maxillae were statistically and clinically insignificant. Six implants were lost prior to loading and one implant has not been restored. No implants or restorations were lost after loading. At 1 year, the implant success rate was 95.6%. Mesiodistal radiographic measurements from 34 patients were averaged, and changes from prosthetic abutment connection to, on average, 12 months follow-up were compared. The radiographs, which were digitalized, measured from the bottom of the implant cylinder to the most coronal bone in contact with implant thread. For mandibular implants, the mean radiographic bone level at prosthetic abutment connection was 1.07 mm; after loading, it was 1.35 mm. For maxillary implants, the mean radiographic bone height at prosthetic abutment connection was 1.16 mm; after loading, it was 1.36 mm. These changes were not statistically significant. The 1-year outcomes from this patient series indicate that one-step Br

  8. Development of a rapid, one-step screening method for the isolation of presumptive proteolytic enterococci.

    Science.gov (United States)

    Graham, Ken; Rea, Rosemary; Simpson, Paul; Stack, Helena

    2017-01-01

    Enterococci show higher proteolytic activities than other lactic acid bacteria and thus have received considerable attention in scientific literature in recent years. Proteolytic enzymes of enterococci have warranted the use of some species as starter, adjuncts or protective cultures and as probiotics, while in some strains they have also been linked with virulence. Consequently, the isolation and identification of proteolytic enterococci is becoming of increasing interest and importance. However, current screening methods for proteolytic enterococci can be time consuming, requiring a two-step procedure which may take up to 96h. This study describes a method, utilising Kanamycin Skim Milk Aesculin Azide (KSMEA) agar, for the isolation of proteolytic enterococci in one-step, thereby significantly reducing screening time. KSMEA combines the selective properties of Kanamycin Aesculin Azide Agar (KAA) with skim milk powder for the detection of proteolytic enterococci. Enterococci produced colonies with a black halo on KSMEA which were accompanied by a zone of clearing in the media when enterococci were proteolytic. KSMEA medium retained the selectivity of KAA, while proteolytic enterococci were easily distinguished from non-proteolytic enterococci when two known strains were propagated on KSMEA. KSMEA also proved effective at isolating and detecting enterococci in raw milk, faeces and soil. Isolates recovered from the screen were confirmed as enterococci using genus-specific primers. Proteolytic enterococci were present in the raw milk sample only and were easily distinguishable from non-proteolytic enterococci and other microorganisms. Therefore, KSMEA provides a rapid, one-step screening method for the isolation of presumptive proteolytic enterococci.

  9. Reversible inactivation of the transcriptional function of P53 protein by farnesylation

    Directory of Open Access Journals (Sweden)

    Berg Danièle

    2006-05-01

    Full Text Available Abstract Background The use of integrating viral vectors in Gene therapy clinical trials has pointed out the problem of the deleterous effect of the integration of the ectopic gene to the cellular genome and the safety of this strategy. We proposed here a way to induce the death of gene modified cells upon request by acting on a pro-apoptotic protein cellular localization and on the activation of its apoptotic function. Results We constructed an adenoviral vector coding a chimeric p53 protein by fusing p53 sequence with the 21 COOH term amino acids sequence of H-Ras. Indeed, the translation products of Ras genes are cytosolic proteins that become secondarily associated with membranes through a series of post-translational modifications initiated by a CAAX motif present at the C terminus of Ras proteins. The chimeric p53HRCaax protein was farnesylated efficiently in transduced human osteosarcoma p53-/- cell line. The farnesylated form of p53 resided mainly in the cytosol, where it is non-functional. Farnesyl transferase inhibitors (FTIs specifically inhibited farnesyl isoprenoid lipid modification of proteins. Following treatment of the cells with an FTI, p53HRCaax underwent translocation into the nucleus where it retained transcription factor activity. Shifting p53 into the nucleus resulted in the induction of p21waf1/CIP1 and Bax transcription, cell growth arrest, caspase activation and apoptosis. Conclusion Artificial protein farnesylation impaired the transcriptional activity of p53. This could be prevented by Farnesyl transferase inhibition. These data highlight the fact that the artificial prenylation of proteins provides a novel system for controlling the function of a transactivating factor.

  10. High rate of mismatch extension during reverse transcription in a single round of retrovirus replication.

    OpenAIRE

    Pulsinelli, G A; Temin, H M

    1994-01-01

    We made spleen necrosis virus-based retroviral vectors with mutations at the 3' end of the primer binding site region to observe the effects of terminal mismatches on retroviral replication. These vectors, when compared to a vector with the wild-type primer binding sequence, allowed us to assay the effects of the mutations on the viral titer during a single cycle of replication. The mutant vectors had titers that were comparable to the wild-type vector, indicating that reverse transcriptase h...

  11. An efficient and more sustainable one-step continuous-flow multicomponent synthesis approach to chromene derivatives

    Science.gov (United States)

    A simple and rapid one-step continuous-flow synthesis route has been developed for the preparation of chromene derivatives from the reaction of aromatic aldehydes, α-cyanomethylene compounds and naphthols. In this contribution, a one-step continuous-flow protocol in a continuous ...

  12. Constructing Competitive Reverse Transcription Polymerize Chain Reaction Inter-Reference of PC mRNA by Intron Approach

    Institute of Scientific and Technical Information of China (English)

    XU Chuang; XIA Cheng; LIU Guo-wen; WANG Zhe; JIANG Yu-fu; ZHANG Nai-sheng; FU Shi-xin

    2004-01-01

    Inter-reference ofcompetitive reverse transcription polymerase chain reaction(RT-PCR)was constructed by intron method to detect the change of PC mRNA level in the pathway of carbohydrate metabolism.The experiment based on the principle that 81bp intron sequence was deleted in PC mRNA compared with PC DNA sequence.The 466bp competitive DNA template recombinant plasmid of PC mRNAwas successfully built by a pair of primer and was cloned once,PC DNA and PC mRNA could be inter-referred each other.The intron approach used in the experiment has broken through the traditional method of constructing competitive template.

  13. Retrotransposon Ty1 RNA contains a 5'-terminal long-range pseudoknot required for efficient reverse transcription.

    Science.gov (United States)

    Huang, Qing; Purzycka, Katarzyna J; Lusvarghi, Sabrina; Li, Donghui; Legrice, Stuart F J; Boeke, Jef D

    2013-03-01

    Ty1 retrotransposon RNA has the potential to fold into a variety of distinct structures, mutation of which affects retrotransposition frequencies. We show here that one potential functional structure is located at the 5' end of the genome and can assume a pseudoknot conformation. Chemoenzymatic probing of wild-type and mutant mini-Ty1 RNAs supports the existence of such a structure, while molecular genetic analyses show that mutations disrupting pseudoknot formation interfere with retrotransposition, indicating that it provides a critical biological function. These defects are enhanced at higher temperatures. When these mutants are combined with compensatory changes, retrotransposition is restored, consistent with pseudoknot architecture. Analyses of mutants suggest a defect in Ty1 reverse transcription. Collectively, our data allow modeling of a three-dimensional structure for this novel critical cis-acting signal of the Ty1 genome.

  14. Simultaneous detection of four garlic viruses by multiplex reverse transcription PCR and their distribution in Indian garlic accessions.

    Science.gov (United States)

    Majumder, S; Baranwal, V K

    2014-06-01

    Indian garlic is infected with Onion yellow dwarf virus (OYDV), Shallot latent virus (SLV), Garlic common latent virus (GarCLV) and allexiviruses. Identity and distribution of garlic viruses in various garlic accessions from different geographical regions of India were investigated. OYDV and allexiviruses were observed in all the garlic accessions, while SLV and GarCLV were observed only in a few accessions. A multiplex reverse transcription (RT)-PCR method was developed for the simultaneous detection and identification of OYDV, SLV, GarCLV and Allexivirus infecting garlic accessions in India. This multiplex protocol standardized in this study will be useful in indexing of garlic viruses and production of virus free seed material.

  15. The sensitivity and specificity of a reverse transcription-polymerase chain reaction assay for the avian pneumovirus (Colorado strain).

    Science.gov (United States)

    Pedersen, J C; Reynolds, D L; Ali, A

    2000-01-01

    A reverse transcription-polymerase chain reaction (RT-PCR) assay for the detection of avian pneumovirus (APV), Colorado strain (US/CO), was evaluated for sensitivity and specificity. The single-tube RT-PCR assay utilized primers developed from the matrix (M) gene sequence of the US/CO APV. The RT-PCR amplified the US/CO APV but did not amplify other pneumoviruses, including the avian pneumoviruses subgroups A and B. The RT-PCR was capable of detecting between 10(0.25) mean tissue culture infective dose (TCID50) and 10(-0.44) TCID50 of the US/CO APV. These results have demonstrated that the single-tube RT-PCR assay is a specific and sensitive assay for the detection of US/CO APV.

  16. Identification and Characterization of Reverse Transcriptase Domain of Transcriptionally Active Retrotransposons in Wheat Genomes

    Institute of Scientific and Technical Information of China (English)

    Yi-Miao TANG; You-Zhi MA; Lian-Cheng LI; Xing-Guo YE

    2005-01-01

    To clarify activation characterization of wheat (Triticum aestivum L.) retrotransposons, transcriptionally active Ty1-copia retrotransposons were found in wheat by using RT-PCR to amplify the RT domain. Sequence analysis of random RT-PCR clones reveals that Ty1-copia retrotransposons are highly heterogeneous and can be divided into at least four groups, which are tentatively named TaRT-1 to TaRT-4.Dot blot hybridization indicates that TaRT- 1 exists in the wheat genome as multiple copies (at 30 000 copies/a hexaploid genome (ABD)). Northern blot hybridization showed that TaRT-1 is only expressed at a low level under normal conditions in seedlings, but at a high level when induced by powdery mildew fungus, jasmonic acid (JA) and salicylic acid (SA). These results suggest that the TaRT-1 expression is highly sensitive to biotic and abiotic stresses.

  17. One-step detection of circulating tumor cells in ovarian cancer using enhanced fluorescent silica nanoparticles

    Directory of Open Access Journals (Sweden)

    Kim JH

    2013-06-01

    Full Text Available Jin Hyun Kim,1,* Hyun Hoon Chung,2,* Min Sook Jeong,1 Mi Ryoung Song,1 Keon Wook Kang,3,4 Jun Sung Kim1 1R&D Center, Biterials Co, Ltd, Seoul, Republic of Korea; 2Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, Republic of Korea; 3Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea; 4Cancer Research Institute, Seoul National University, Seoul, Republic of Korea *These authors contributed equally to this work Abstract: Ovarian cancer is the fifth-leading cause of cancer-related deaths among women as a result of late diagnosis. For survival rates to improve, more sensitive and specific methods for earlier detection of ovarian cancer are needed. This study presents the development of rapid and specific one-step circulating tumor cell (CTC detection using flow cytometry in a whole-blood sample with fluorescent silica nanoparticles. We prepared magnetic nanoparticle (MNP-SiO2(rhodamine B isothiocyanate [RITC] (MNP-SiO2[RITC] incorporating organic dyes [RITC, λmax(ex/em = 543/580 nm] in the silica shell. We then controlled the amount of organic dye in the silica shell of MNP-SiO2(RITC for increased fluorescence intensity to overcome the autofluorescence of whole blood and increase the sensitivity of CTC detection in whole blood. Next, we modified the surface function group of MNP-SiO2(RITC from –OH to polyethylene glycol (PEG/COOH and conjugated a mucin 1 cell surface-associated (MUC1 antibody on the surface of MNP-SiO2(RITC for CTC detection. To study the specific targeting efficiency of MUC1-MNP-SiO2(RITC, we used immunocytochemistry with a MUC1-positive human ovarian cancer cell line and a negative human embryonic kidney cell line. This technology was capable of detecting 100 ovarian cancer cells in 50 µL of whole blood. In conclusion, we developed a one-step CTC detection technology in ovarian cancer based on multifunctional silica nanoparticles

  18. One-step preparation and photocatalytic performance of vanadium doped TiO{sub 2} coatings

    Energy Technology Data Exchange (ETDEWEB)

    Vasilić, R., E-mail: rastko.vasilic@ff.bg.ac.rs [University of Belgrade, Faculty of Physics, Studentski trg 12-16, 11000 Belgrade (Serbia); Stojadinović, S. [University of Belgrade, Faculty of Physics, Studentski trg 12-16, 11000 Belgrade (Serbia); Radić, N. [University of Belgrade, Institute of Chemistry, Technology and Metallurgy, Department of Catalysis and Chemical Engineering, Njegoševa 12, 11000 Belgrade (Serbia); Stefanov, P. [Institute of General and Inorganic Chemistry, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Block 11, Sofia 1113 (Bulgaria); Dohčević-Mitrović, Z. [University of Belgrade, Institute of Physics, Pregrevica 118, 11080 Belgrade (Serbia); Grbić, B. [University of Belgrade, Institute of Chemistry, Technology and Metallurgy, Department of Catalysis and Chemical Engineering, Njegoševa 12, 11000 Belgrade (Serbia)

    2015-02-01

    In this paper, we have investigated one-step preparation of vanadium doped TiO{sub 2} coatings formed by plasma electrolytic oxidation (PEO) of titanium in electrolyte containing 10 g/L Na{sub 3}PO{sub 4}·12H{sub 2}O + 0.5 g/L NH{sub 4}VO{sub 3}. The morphology, phase structure, and elemental composition of the formed coatings were characterized by atomic force microscopy (AFM), x-ray diffraction (XRD), and x-ray photoelectron spectroscopy (XPS) techniques. Ultraviolet–visible diffuse reflectance spectroscopy (UV–Vis DRS) was employed to evaluate the band gap energy of obtained coatings. Vanadium doped TiO{sub 2} coatings are partly crystallized and mainly composed of anatase phase TiO{sub 2}, with up to about 2 wt% of vanadium present in the surface layer of the oxide. The valence band photoelectron spectra and UV–Vis DRS showed that vanadium doped TiO{sub 2} coatings exhibit notable red shift with respect to the pure TiO{sub 2} coatings. The photocatalytic activity was evaluated by monitoring the degradation of methyl orange under simulated sunlight conditions. Photocatalytic activity of vanadium doped TiO{sub 2} coatings increases with PEO time. Prolonged PEO times result in higher roughness of obtained coatings, thus increasing surface area available for methyl orange degradation. Vanadium doped TiO{sub 2} coatings obtained after 180 s of PEO time exhibit the best photocatalytic activity and about 67% of methyl orange is degraded after 12 h of irradiation under simulated sunlight. - Highlights: • One-step preparation of V-doped TiO{sub 2} coatings in 10 g/L Na{sub 3}PO{sub 4}·12H{sub 2}O + 0.5 g/L NH{sub 4}VO{sub 3}. • Properties of obtained coatings strongly depend on microdischarge characteristics. • Band gap of V-doped TiO{sub 2} coatings is shifted towards red side of the spectrum. • V-doped TiO{sub 2} coatings have better photocatalytic activity than pure TiO{sub 2}. • After 12 h of simulated sunlight irradiation, 67% of

  19. H2-Assistance One-Step Growth of Si Nanowires and Their Growth Mechanism

    Institute of Scientific and Technical Information of China (English)

    QIU Ming-Xia; RUAN Shuang-Chen; GAO Biao; HUO Kai-Fu; ZHAI Jian-Pang; LI Ling; LIAO Hui; XU Xin-Tong

    2011-01-01

    Large-scale nanowires are grown on Si wafers by the catalyst-free one-step thermal reaction method in Ar/H2 mixture atmosphere at 1000℃. The x-ray diffraction and energy dispersive x-ray spectroscopy results reveal that the final nanowires are of silicon nanostructures. The Held emission scanning electron microscopy shows that these self-organized Si nanowires (SiNWs) possess curly crowns with diameters varying from 10 to 300 nm and lengths of up to several hundreds of micrometers. The transmission electron microscopy indicates that the nanowires are pure Si with amorphous structures. All the measurement results show that no silicon oxide is generated in our products. The growth mechanism is proposed tentatively. Silicon oxide is reduced into Si nanoparticles under the Ar/H2 mixture, which is the main reason for the formation of such SiNWs. Our experiments offer a method of preparing Si nanostructures by simply reducing silicon oxide at high temperature.%Large-scale nanowires are grown on Si wafers by the catalyst-free one-step thermal reaction method in Ar/H2 mixture atmosphere at 1000℃.The x-ray diffraction and energy dispersive x-ray spectroscopy results reveal that the final nanowires are of silicon nanostructures.The field emission scanning electron microscopy shows that these self-organized Si nanowires (SiNWs) possess curly crowns with diameters varying from 10 to 300nm and lengths of up to several hundreds of micrometers.The transmission electron microscopy indicates that the nanowires are pure Si with amorphous structures.All the measurement results show that no silicon oxide is generated in our products.The growth mechanism is proposed tentatively.Silicon oxide is reduced into Si nanoparticles under the Ar/H2 mixture,which is the main reason for the formation of such SiNWs.Our experiments offer a method of preparing Si nanostructures by simply reducing silicon oxide at high temperature.Silicon nanowires (SiNWs) have higher carrier mobility,a larger

  20. Relationship between mechanical properties of one-step self-etch adhesives and water sorption.

    Science.gov (United States)

    Hosaka, Keiichi; Nakajima, Masatoshi; Takahashi, Masahiro; Itoh, Shima; Ikeda, Masaomi; Tagami, Junji; Pashley, David H

    2010-04-01

    The purpose of this study was to evaluate the relationship between changes in the modulus of elasticity and ultimate tensile strength of one-step self-etch adhesives, and their degree of water sorption. Five one-step self-etch adhesives, Xeno IV (Dentsply Caulk), G Bond (GC Corp.), Clearfil S3 Bond (Kuraray Medical Inc.), Bond Force (Tokuyama Dental Corp.), and One-Up Bond F Plus (Tokuyama Dental Corp.) were used. Ten dumbelled-shaped polymers of each adhesive were used to obtain the modulus of elasticity by the three-point flexural bending test and the ultimate tensile strength by microtensile testing. The modulus of elasticity and the ultimate tensile strength were measured in both dry and wet conditions before/after immersion in water for 24h. Water sorption was measured, using a modification of the ISO-4049 standard. Each result of the modulus of elasticity and ultimate tensile strength was statistically analyzed using a two-way ANOVA and the result of water sorption was statistically analyzed using a one-way ANOVA. Regression analyses were used to determine the correlations between the modulus of elasticity and the ultimate tensile strength in dry or wet states, and also the percent decrease in these properties before/after immersion of water vs. water sorption. In the dry state, the moduli of elasticity of the five adhesive polymers varied from 948 to 1530 MPa, while the ultimate tensile strengths varied from 24.4 to 61.5 MPa. The wet specimens gave much lower moduli of elasticity (from 584 to 1073 MPa) and ultimate tensile strengths (from 16.5 to 35.0 MPa). Water sorption varied from 32.1 to 105.8 g mm(-3). The moduli of elasticity and ultimate tensile strengths of the adhesives fell significantly after water-storage. Water sorption depended on the constituents of the adhesive systems. The percent decreases in the ultimate tensile strengths of the adhesives were related to water sorption, while the percent reductions in the moduli of elasticity of the

  1. External Quality Assessment for the Detection of Measles Virus by Reverse Transcription-PCR Using Armored RNA.

    Directory of Open Access Journals (Sweden)

    Dong Zhang

    Full Text Available In recent years, nucleic acid tests for detection of measles virus RNA have been widely applied in laboratories belonging to the measles surveillance system of China. An external quality assessment program was established by the National Center for Clinical Laboratories to evaluate the performance of nucleic acid tests for measles virus. The external quality assessment panel, which consisted of 10 specimens, was prepared using armored RNAs, complex of noninfectious MS2 bacteriophage coat proteins encapsulated RNA of measles virus, as measles virus surrogate controls. Conserved sequences amplified from a circulating measles virus strain or from a vaccine strain were encapsulated into these armored RNAs. Forty-one participating laboratories from 15 provinces, municipalities, or autonomous regions that currently conduct molecular detection of measles virus enrolled in the external quality assessment program, including 40 measles surveillance system laboratories and one diagnostic reagent manufacturer. Forty laboratories used commercial reverse transcription-quantitative PCR kits, with only one laboratory applying a conventional PCR method developed in-house. The results indicated that most of the participants (38/41, 92.7% were able to accurately detect the panel with 100% sensitivity and 100% specificity. Although a wide range of commercially available kits for nucleic acid extraction and reverse transcription polymerase chain reaction were used by the participants, only two false-negative results and one false-positive result were generated; these were generated by three separate laboratories. Both false-negative results were obtained with tests performed on specimens with the lowest concentration (1.2 × 104 genomic equivalents/mL. In addition, all 18 participants from Beijing achieved 100% sensitivity and 100% specificity. Overall, we conclude that the majority of the laboratories evaluated have reliable diagnostic capacities for the detection

  2. Rapid one-step assays for on-site monitoring of mouse and rat urinary allergens.

    Science.gov (United States)

    Koets, Marjo; Renström, Anne; Zahradnik, Eva; Bogdanovic, Jelena; Wouters, Inge M; van Amerongen, Aart

    2011-12-01

    Allergy to rodent proteins is common among laboratory animal workers. Sensitive methods to measure exposure to these allergens have been developed. These assays are, however, expensive, time-consuming, and require a laboratory facility and methodological expertise. A simple method to screen for allergen spread, or to test whether hygiene standards are maintained, would be useful. Lateral flow immunoassays (LFIAs) are especially suited for field settings; the tests are simple and results are visible within minutes. LFIAs were developed for detection of the rodent urinary allergens Mus m 1 and Rat n 1. Pilot studies were performed in animal facilities in three countries using both extracts from airborne dust samples and samples collected by wiping surfaces. For comparison and determination of sensitivity, the concentrations of rodent urinary allergens in the samples were also measured using enzyme immunoassays (EIAs). The LFIAs for rat and mouse urinary allergens had a detection limit of 31 pg allergen per mL in a buffer system with purified allergen standards. Results of environmental dust extracts tested in LFIAs correlated well with levels obtained using EIAs. Spread of rodent allergens, or non-adherence to hygiene around laboratory animal facilities, may aggravate rodent allergy. Using a simple, sensitive one-step assay, allergens can be detected to prevent allergen exposure. The results reveal that the rapid assays are suited for on-site demonstration of exposure to rodent allergens, and thus, useful in occupational hygiene practice.

  3. A one-step-ahead pseudo-DIC for comparison of Bayesian state-space models.

    Science.gov (United States)

    Millar, R B; McKechnie, S

    2014-12-01

    In the context of state-space modeling, conventional usage of the deviance information criterion (DIC) evaluates the ability of the model to predict an observation at time t given the underlying state at time t. Motivated by the failure of conventional DIC to clearly choose between competing multivariate nonlinear Bayesian state-space models for coho salmon population dynamics, and the computational challenge of alternatives, this work proposes a one-step-ahead DIC, DICp, where prediction is conditional on the state at the previous time point. Simulations revealed that DICp worked well for choosing between state-space models with different process or observation equations. In contrast, conventional DIC could be grossly misleading, with a strong preference for the wrong model. This can be explained by its failure to account for inflated estimates of process error arising from the model mis-specification. DICp is not based on a true conditional likelihood, but is shown to have interpretation as a pseudo-DIC in which the compensatory behavior of the inflated process errors is eliminated. It can be easily calculated using the DIC monitors within popular BUGS software when the process and observation equations are conjugate. The improved performance of DICp is demonstrated by application to the multi-stage modeling of coho salmon abundance in Lobster Creek, Oregon. © 2014, The International Biometric Society.

  4. One-Step Electrodeposited Nickel Cobalt Sulfide Nanosheet Arrays for High-Performance Asymmetric Supercapacitors

    KAUST Repository

    Chen, Wei

    2014-09-23

    A facile one-step electrodeposition method is developed to prepare ternary nickel cobalt sulfide interconnected nanosheet arrays on conductive carbon substrates as electrodes for supercapacitors, resulting in exceptional energy storage performance. Taking advantages of the highly conductive, mesoporous nature of the nanosheets and open framework of the three-dimensional nanoarchitectures, the ternary sulfide electrodes exhibit high specific capacitance (1418 F g(-1) at 5 A g(-1) and 1285 F g(-1) at 100 A g(-1)) with excellent rate capability. An asymmetric supercapacitor fabricated by the ternary sulfide nanosheet arrays as positive electrode and porous graphene film as negative electrode demonstrates outstanding electrochemical performance for practical energy storage applications. Our asymmetric supercapacitors show a high energy density of 60 Wh kg(-1) at a power density of 1.8 kW kg(-1). Even when charging the cell within 4.5 s, the energy density is still as high as 33 Wh kg(-1) at an outstanding power density of 28.8 kW kg(-1) with robust long-term cycling stability up to 50 000 cycles.

  5. One-step large-scale deposition of salt-free DNA origami nanostructures

    Science.gov (United States)

    Linko, Veikko; Shen, Boxuan; Tapio, Kosti; Toppari, J. Jussi; Kostiainen, Mauri A.; Tuukkanen, Sampo

    2015-10-01

    DNA origami nanostructures have tremendous potential to serve as versatile platforms in self-assembly -based nanofabrication and in highly parallel nanoscale patterning. However, uniform deposition and reliable anchoring of DNA nanostructures often requires specific conditions, such as pre-treatment of the chosen substrate or a fine-tuned salt concentration for the deposition buffer. In addition, currently available deposition techniques are suitable merely for small scales. In this article, we exploit a spray-coating technique in order to resolve the aforementioned issues in the deposition of different 2D and 3D DNA origami nanostructures. We show that purified DNA origamis can be controllably deposited on silicon and glass substrates by the proposed method. The results are verified using either atomic force microscopy or fluorescence microscopy depending on the shape of the DNA origami. DNA origamis are successfully deposited onto untreated substrates with surface coverage of about 4 objects/mm2. Further, the DNA nanostructures maintain their shape even if the salt residues are removed from the DNA origami fabrication buffer after the folding procedure. We believe that the presented one-step spray-coating method will find use in various fields of material sciences, especially in the development of DNA biochips and in the fabrication of metamaterials and plasmonic devices through DNA metallisation.

  6. One-step uniform growth of magnesium hydride nanoparticles on graphene

    Directory of Open Access Journals (Sweden)

    Yuqin Huang

    2017-02-01

    Full Text Available A simple solvent-free method to synthesize MgH2 nanoparticles (MgH2 NPs uniformly grown on graphene nanosheets (GNs has been reported in this paper. Based on the formation of MgH2 by di-n-butylmagnesium ((C4H92Mg thermal decomposition under hydrogen pressure, the GNs were added as matrix to hinder the agglomeration and growing of MgH2 NPs. The fabricated MgH2/GNs nanocomposites, in which MgH2 NPs were homogenously growing in the graphene matrix, have been synthesized by the favorable adsorption energy between (C4H92Mg and GNs. Resulting from the one-step solvent-free route, the generated MgH2 NPs shows high hydrogen capacity and steady hydriding and dehydriding properties, without the interference of the synthetic medium. At the same time, the size of the fabricated MgH2 NPs can be controlled by adjusting the mass ratio of MgH2 to graphene, the various hydrogen pressure and temperature. Attributed to smaller size effect, well uniform distribution of high density MgH2 NPs, and the agglomeration blocking ability of graphene, the MgH2/GNs-40 wt% exhibits the favorite hydrogen storage performance.

  7. Perovskite Hollow Fibers with Precisely Controlled Cation Stoichiometry via One-Step Thermal Processing.

    Science.gov (United States)

    Zhu, Jiawei; Zhang, Guangru; Liu, Gongping; Liu, Zhengkun; Jin, Wanqin; Xu, Nanping

    2017-05-01

    The practical applications of perovskite hollow fibers (HFs) are limited by challenges in producing these easily, cheaply, and reliably. Here, a one-step thermal processing approach is reported for the efficient production of high performance perovskite HFs, with precise control over their cation stoichiometry. In contrast to traditional production methods, this approach directly uses earth-abundant raw chemicals in a single thermal process. This approach can control cation stoichiometry by avoiding interactions between the perovskites and polar solvents/nonsolvents, optimizes sintering, and results in high performance HFs. Furthermore, this method saves much time and energy (≈ 50%), therefore pollutant emissions are greatly reduced. One successful example is Ba0.5Sr0.5Co0.8Fe0.2O3-δ HFs, which are used in an oxygen-permeable membrane. This exhibits high oxygen permeation flux values that exceed desired commercial targets and compares favorably with previously reported oxygen-permeable membranes. Studies on other perovskites have produced similarly successful results. Overall, this approach could lead to energy efficient, solid-state devices for industrial application in energy and environmental fields. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Self-Stratified Antimicrobial Acrylic Coatings via One-Step UV Curing.

    Science.gov (United States)

    Zhao, Jie; Millians, William; Tang, Saide; Wu, Tiehang; Zhu, Lei; Ming, Weihua

    2015-08-26

    We designed and synthesized a novel quaternary ammonium methacrylate compound (QAC-2) bearing a perfluoroalkyl tail on one end and an acrylic moiety on the other. Via one-step UV curing of QAC-2 and methyl methacrylate (MMA) with ethylene glycol dimethacrylate (EGDMA) as the cross-linker, we obtained cross-linked coatings with excellent antimicrobial property, as demonstrated by the total kill against both Gram-negative Escherichia coli (E. coli) and Gram-positive Staphylococcus epidermidis (S. epidermidis) at a QAC-2 concentration as low as ∼0.06 mol % (∼0.4 wt %) relative to MMA, which was substantially lower than the QAC amount needed in the coatings containing QACs with a hydrocarbon tail. A zone of inhibition test confirmed that the antimicrobial effect was on the basis of contact killing and there was no leaching of antimicrobial species from the cross-linked coating. The high antimicrobial potency in QAC-2-containing films was the consequence of strong surface enrichment of the fluorinated QAC, as confirmed by X-ray photoelectron spectroscopy (XPS).

  9. Preparation of native cellulose-AgCl fiber with antimicrobial activity through one-step electrospinning.

    Science.gov (United States)

    Wang, Shaojun; Zhang, Xiaomin; Luo, Ting; Zhu, Jin; Su, Shengpei

    2017-02-01

    The native Cellulose-AgCl fiber have been firstly fabricated by one-step electrospinning of cellulose solution with poly(vinyl pyrrolidone) (PVP) and AgNO3. X-ray diffraction, Scanning electron microscopy (SEM), Energy dispersive spectrometer, Thermo-gravimetric analysis and Fourier transform infrared are used to characterize the crystal structure, morphology and composition of cellulose-AgCl nanocomposites. The results of SEM indicate that the size of AgCl in cellulose fiber matrix is able to be adjusted by the addition of Polyvinylpyrrolidone (PVP). The antimicrobial activity of the nanocomposites fiber is also tested against the model microbes E. coli (Gram-negative) and S. aureus (Gram-positive). The results indicate that cellulose-AgCl nanocomposites have a good antimicrobial activity, which is improving with the decrease of AgCl size in fiber matrix. This work provides a novel and simple way to adjust the AgCl size in electrospinning cellulose matrix which can be applied as functional biomaterials.

  10. One-step synthesis of boronic acid functionalized gold nanoclusters for photoluminescence sensing of dopamine

    Science.gov (United States)

    Chen, Huide; Liu, Chunxiu; Xia, Yunsheng

    2017-03-01

    This study is the first to report one-step synthesis of boronic acid functionalized gold nanoclusters (AuNCs) using mixed ligands of 4-mercaptophenylboronic acid (MPBA) and glutathione. Furthermore, the emission color of the products can be fancily tuned from green to near-infrared by simply changing the proportion of the two stabilizers. In basic media, dopamine (DA) molecules themselves polymerize each other and form polydopamine with large amounts of cis-diol groups, which then react with boronic acid groups on the AuNC’s surface based on the formation of boronate esters. As a result, the photoluminescence of the AuNCs is well quenched by the electron transfer effect. Accordingly, DA molecules are assayed from 0.5 to 9 μM, and the detection limit is as low as 0.1 μM. The as-prepared AuNCs exhibit high selectivity; the existing biomolecules including various amino acids, ascorbic acid, uric acid, glucose, etc, do not interfere with the assay. The proposed method is successfully applied to the assay of DA in human serum, indicating its practical potential.

  11. One-Step Green Synthesis of Metallic Nanoparticles Using Sodium Alginate

    Directory of Open Access Journals (Sweden)

    Jesus Valdez

    2016-01-01

    Full Text Available Metallic nanoparticles have been focus of research because of their characteristic properties, specifically the LSPR which can have wide applications in biomedical sciences and engineering. Currently, traditional physical and chemical methods can synthesize these nanoparticles but their disadvantages such as costs, time, effectiveness, and toxicity of precursors provide a wide range of problems for the synthesis of these nanoparticles. Recently, some natural polymers and organic compounds have been used for the synthesis of nanoparticles by green methods. In this study, we synthesize copper, silver, and gold nanoparticles using sodium alginate as reducing and stabilizing agent under microwave irradiation. The LSPR for each system was observed by UV-vis spectroscopy. Particle size distribution and zeta potential demonstrate the size and stability for these nanoparticles. FESEM and TEM microscopies have shown the size and morphology of these systems correlated with UV-vis, particle size distribution, and zeta potential analyses. The study demonstrates a rapid, facile, cheaper, and one-step green method of synthesis for these metallic nanoparticles being an alternative to the conventional methods used for synthesis of metallic nanoparticles.

  12. One-step femtosecond laser welding and internal machining of three glass substrates

    Science.gov (United States)

    Tan, Hua; Duan, Ji'an

    2017-05-01

    In this paper, it demonstrated one-step femtosecond laser welding and internal machining of three fused silica substrates in the optical- and non-optical-contact regimes by focusing 1030-nm laser pulses at the middle of the second substrate. Focusing laser pulses within the second glass in optical-contact and non-optical-contact samples induces permanent internal structural modification, leading to the three glass substrates bonding together simultaneously. The bonding mechanism is based on the internal modification of glass, and this mechanism is different from that of ordinary glass welding at the interface. Welding-spot size is affected by not only the gap distance (ablation effect) and heat transmission, but also by gravity through examining the sizes of the welding spots on the four contact welding surfaces. The maximum bonding strength of the lower interface (56.2 MPa) in the optical-contact regime is more than double that (27.6 MPa) in the non-optical-contact regime.

  13. One-step Fabrication of Nanoporous Black Silicon Surfaces for Solar Cells using Modified Etching Solution

    Institute of Scientific and Technical Information of China (English)

    Ye-hua Tang; Chun-lan Zhou; Su Zhou; Yan Zhao; Wen-jing Wang; Jian-ming Fei; Hong-bin Cao

    2013-01-01

    Currently,a conventional two-step method has been used to generate black silicon (BS)surfaces on silicon substrates for solar cell manufacturing.However,the performances of the solar cell made with such surface generation method are poor,because of the high surface recombination caused by deep etching in the conventional surface generation method for BS.In this work,a modified wet chemical etching solution with additives was developed.A zhomogeneous BS layer with random porous structure was obtained from the modified solution in only one step at room temperature.The BS layer had low reflectivity and shallow etching depth.The additive in the etch solution performs the function of pH-modulation.After 16-min etching,the etching depth in the samples was approximately 200 nm,and the spectrum-weighted-reflectivity in the range from 300 nm to 1200 nm was below 5%,BS solar cells were fabricated in the production line.The decreased etching depth can improve the electrical performance of solar cells because of the decrease in surface recombination.An efficiency of 15,63% for the modified etching BS solar cells was achieved on a large area,ptype single crystalline silicon substrate with a 624.32-mV open circuit voltage and a 77.88%fill factor.

  14. Detection of Zika virus by SYBR green one-step real-time RT-PCR.

    Science.gov (United States)

    Xu, Ming-Yue; Liu, Si-Qing; Deng, Cheng-Lin; Zhang, Qiu-Yan; Zhang, Bo

    2016-10-01

    The ongoing Zika virus (ZIKV) outbreak has rapidly spread to new areas of Americas, which were the first transmissions outside its traditional endemic areas in Africa and Asia. Due to the link with newborn defects and neurological disorder, numerous infected cases throughout the world and various mosquito vectors, the virus has been considered to be an international public health emergency. In the present study, we developed a SYBR Green based one-step real-time RT-PCR assay for rapid detection of ZIKV. Our results revealed that the real-time assay is highly specific and sensitive in detection of ZIKV in cell samples. Importantly, the replication of ZIKV at different time points in infected cells could be rapidly monitored by the real-time RT-PCR assay. Specifically, the real-time RT-PCR showed acceptable performance in measurement of infectious ZIKV RNA. This assay could detect ZIKV at a titer as low as 1PFU/mL. The real-time RT-PCR assay could be a useful tool for further virology surveillance and diagnosis of ZIKV.

  15. Adsorption of phosphate in water using one-step synthesized zirconium-loaded reduced graphene oxide

    Science.gov (United States)

    Luo, Xin; Wang, Xiurong; Bao, Shaopan; Liu, Xiawei; Zhang, Weicheng; Fang, Tao

    2016-12-01

    In this account, a one-step green hydrothermal method for zirconium-loaded reduced graphene oxide (RGO-Zr) adsorbent was developed in pure water. It is based on the formation of initially strong-coupling RGO-Zr nanocomposites followed by in situ reduction of GO to RGO during the hydrothermal treatment. The phosphate adsorption performance of the as-prepared nanocomposites was investigated in aqueous environment under various conditions. The characterization results of RGO-Zr nanocomposites showed that ZrO2 was successfully integrated onto the RGO sheets in amorphous. The data from equilibrium phosphate adsorption on RGO-Zr revealed that the adsorption kinetics followed a pseudo-second-order kinetic model, where the adsorption isotherm fitted the Langmuir isotherm model with a maximum adsorption capacity of 27.71 mg P/g at pH 5 and 298 K. The improved phosphate adsorption on RGO-Zr was caused by the dispersion of ZrO2 on the RGO surface. Furthermore, the phosphate adsorption was found insensitive to the increase in pH while it was sensitive to the increase in temperature. The coexisting anions of SO42-, F-, Cl-, NO3- and CO32- affected the phosphate adsorption in a different way. Results suggest that the present RGO-Zr adsorbent has the potential for controlling phosphorus pollution in water.

  16. One-step electrochemical synthesis of graphene/polyaniline composite film and its applications

    Energy Technology Data Exchange (ETDEWEB)

    Feng, Xiao-Miao; Li, Rui-Mei; Ma, Yan-Wen; Chen, Run-Feng; Shi, Nai-En; Fan, Qu-Li; Huang, Wei [Key Laboratory for Organic Electronics and Information Displays (KLOEID) and Institute of Advanced Materials (IAM), Nanjing University of Posts and Telecommunications (NUPT), Nanjing 210046 (China)

    2011-08-09

    This work describes a new one-step large-scale electrochemical synthesis of graphene/polyaniline (PANI) composite films using graphite oxide (GO) and aniline as the starting materials. The size of the film could be controlled by the area of indium tin oxide (ITO). Scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), and ultraviolet-visible absorption spectrum (UV-vis) results demonstrated that the graphene/PANI composite film was successfully synthesized. The obtained graphene/PANI composite film showed large specific area, high conductivity, good biocompatibility, and fast redox properties and had perfect layered and encapsulated structures. Electrochemical experiments indicated that the composite film had high performances and could be widely used in applied electrochemical fields. As a model, horseradish peroxidase (HRP) was entrapped onto the film-modified glassy carbon electrode (GCE) and used to construct a biosensor. The immobilized HRP showed a pair of well-defined redox peaks and high catalytic activity for the reduction of H{sub 2}O{sub 2}. Furthermore, the graphene/PANI composite film could be directly used as the supercapacitor electrode. The supercapacitor showed a high specific capacitance of 640 F g{sup -1} with a retention life of 90% after 1000 charge/discharge cycles. (Copyright copyright 2011 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  17. One-step process for superhydrophobic metallic surfaces by wire electrical discharge machining.

    Science.gov (United States)

    Bae, Won Gyu; Song, Ki Young; Rahmawan, Yudi; Chu, Chong Nam; Kim, Dookon; Chung, Do Kwan; Suh, Kahp Y

    2012-07-25

    We present a direct one-step method to fabricate dual-scale superhydrophobic metallic surfaces using wire electrical discharge machining (WEDM). A dual-scale structure was spontaneously formed by the nature of exfoliation characteristic of Al 7075 alloy surface during WEDM process. A primary microscale sinusoidal pattern was formed via a programmed WEDM process, with the wavelength in the range of 200 to 500 μm. Notably, a secondary roughness in the form of microcraters (average roughness, Ra: 4.16 to 0.41 μm) was generated during the exfoliation process without additional chemical treatment. The low surface energy of Al 7075 alloy (γ = 30.65 mJ/m(2)) together with the presence of dual-scale structures appears to contribute to the observed superhydrophobicity with a static contact angle of 156° and a hysteresis less than 3°. To explain the wetting characteristics on dual-scale structures, we used a simple theoretical model. It was found that Cassie state is likely to present on the secondary roughness in all fabricated surfaces. On the other hand, either Wenzel or Cassie state can present on the primary roughness depending on the characteristic length of sinusoidal pattern. In an optimal condition of the serial cutting steps with applied powers of ∼30 and ∼8 kW, respectively, a stable, superhydrophobic metallic surface was created with a sinusoidal pattern of 500 μm wavelength.

  18. One-Step Electrochemical Preparation of Multilayer Graphene Functionalized with Nitrogen

    Science.gov (United States)

    Ustavytska, Olena; Kurys, Yaroslav; Koshechko, Vyacheslav; Pokhodenko, Vitaly

    2017-03-01

    A new environmentally friendly one-step method for producing multilayer (preferably 7-9 layers) nitrogen-doped graphene (N-MLG) with a slight amount of oxygen-containing defects was developed. The approach is based on the electrochemical exfoliation of graphite electrode in the presence of azide ions under the conditions of electrolysis with pulse changing of the electrode polarization potential. It was found that usage of azide anions lead not only to the exfoliation of graphite but also to the simultaneous functionalization of graphene sheets by nitrogen atoms (as a result of electrochemical decomposition of azide anions with ammonia evolution). Composition, morphology, structure, and electrochemical properties of N-MLG were characterized by C,H,N analysis, transmission electron microscopy, atomic force microscopy, FTIR, UV-Vis, and Raman spectroscopy, as well as cyclic voltammetry. The perspective of using N-MLG as oxygen reduction reaction electrocatalyst and for the electrochemical analysis of biomarkers (dopamine, ascorbic acid, and uric acid) in their mixtures was shown.

  19. One-step solid state synthesis of capped γ-Fe(2)O(3) nanocrystallites.

    Science.gov (United States)

    Zboril, R; Bakandritsos, A; Mashlan, M; Tzitzios, V; Dallas, P; Trapalis, Ch; Petridis, D

    2008-03-05

    The thermally induced solid state synthesis of soluble organophilic maghemite (γ-Fe(2)O(3)) nanocrystallites is described. The solvent-free one-step synthesis involves the reaction in the melt state of Fe(NO)(3)·9H(2)O and RCOOH (R = C(11)H(23), C(15)H(31)) at 240 °C. The method yields well-crystallized nanoparticles of γ-Fe(2)O(3) functionalized with the corresponding aliphatic acid. Transmission electron microscopy (TEM) and atomic force microscopy (AFM) observations reveal composite particles with faceted magnetic cores and average size of 20 nm, which are well capped with the surrounding organic sheath. The Fourier transform infrared (FT-IR) spectra and thermal analysis suggest a bimodal configuration of the organic shell including chemically coordinated and physisorbed molecules of aliphatic acid. The chemical bonding of the carboxylate groups to the surface iron atoms is also indicated by a paramagnetic doublet with unchanged area in the variable temperature Mössbauer spectra. The spinel γ-Fe(2)O(3) particles exhibit perfect structural and magnetic ordering, including the almost ideal ratio of octahedral to tetrahedral positions (5/3) and very low degree of spin canting, as confirmed by in-field Mössbauer spectroscopy. Magnetic measurements demonstrate the suitable properties required in various (bio)magnetic applications like superparamagnetic behavior at room temperature, high saturation magnetization achievable at low applied fields and suppressed magnetic interactions.

  20. One-step generation of error-prone PCR libraries using Gateway® technology

    Directory of Open Access Journals (Sweden)

    Gruet Antoine

    2012-01-01

    Full Text Available Abstract Background Error-prone PCR (epPCR libraries are one of the tools used in directed evolution. The Gateway® technology allows constructing epPCR libraries virtually devoid of any background (i.e., of insert-free plasmid, but requires two steps: the BP and the LR reactions and the associated E. coli cell transformations and plasmid purifications. Results We describe a method for making epPCR libraries in Gateway® plasmids using an LR reaction without intermediate BP reaction. We also describe a BP-free and LR-free sub-cloning method for in-frame transferring the coding sequence of selected clones from the plasmid used to screen the library to another one devoid of tag used for screening (such as the green fluorescent protein. We report preliminary results of a directed evolution program using this method. Conclusions The one-step method enables producing epPCR libraries of as high complexity and quality as does the regular, two-step, protocol for half the amount of work. In addition, it contributes to preserve the original complexity of the epPCR product.

  1. A rapid one-step electrodeposition process for fabrication of superhydrobic surfaces on anode and cathode

    Institute of Scientific and Technical Information of China (English)

    郝丽梅; 闫小乐; 解忧; 张涛; 陈志

    2016-01-01

    This work presents a method to solve the weak solubility of zinc chloride (ZnCl2) in the ethanol by adding some reasonable water into an ethanol electrolyte containing ZnCl2and myristic acid (CH3(CH2)12COOH). A rapid one-step electrodeposition process was developed to fabricate anodic (2.5 min) and cathodic (40 s) superhydrophobic surfaces of copper substrate (contact angle more than 150°) in an aqueous ethanol electrolyte. Morphology, composition, chemical structure and superhydrophobicity of these superhydrophobic surfaces were investigated by SEM, FTIR, XRD, and contact angle measurement, respectively. The results indicate that water ratio of the electrolyte can reduce the required deposition time, superhydrophobic surface needs over 30 min with anhydrous electrolyte, while it needs only 2.5 min with electrolyte including 10 mL water, and the maximum contact angle of anodic surface is 166° and that of the cathodic surface is 168°. Two copper electrode surfaces have different reactions in the process of electrodeposition time, and the anodic copper surface covers copper myristate (Cu[CH3(CH2)12COO]2) and cupric chloride (CuCl); while, zinc myristate (Zn[CH3(CH2)12COO]2) and pure zinc (Zn) appear on the cathodic surface.

  2. A novel oromucosal prolonged release mucoadhesive suspension by one step spray coagulation method.

    Science.gov (United States)

    Cilurzo, Francesco; Gennari, Chiara G M; Selmin, Francesca; Musazzi, Umberto M; Rumio, Cristiano; Minghetti, Paola

    2013-06-01

    An oromucosal mucoadhesive suspension (OMS) able to combine the peculiarities of prolonged release mucoadhesive microparticles with those of an immediate release oromucosal solution is described. Microparticles were obtained by ionotropic gelation of alginate blended with another mucoadhesive material in a one step process where the cross-linking bath constituted the suspension vehicle. The effects of formulation and processing conditions on OMS performances were measured in-vitro determining the enhancement of drug penetration in buccal porcine mucosa and inhibition of tooth plaque formation using flurbiprofen and delmopinol as model drugs, respectively. Well-formed and spherical microparticles were obtained combining alginate with carbomer; linear dependence of particle size from the feed composition, viscosity and atomization pressure was found. As demonstrated by using FITC-labelled microparticles, the system remained onto the buccal mucosa at least for a six hour period. As a consequence, 0.1% flurbiprofen OMS guaranteed a concentration of flurbiprofen into buccal porcine mucosa over 6 hours comparable to 0.25% flurbiprofen reference solution, allowing a potential reduction of the 60% administered dose. The use of in-house made artificial mouth revealed that the once-a-day administration of 0.1% delmopinol OMS was as effective in plaque inhibition as the 0.2% delmopinol reference solution product given twice-a-day. These results suggested that the development of bioadhesive oromucosal suspensions, localizing the drug into buccal cavity, can reduce regimen and administrated dose.

  3. One-Step Synthesis of PEGylated Gold Nanoparticles with Tunable Surface Charge

    Directory of Open Access Journals (Sweden)

    Rares Stiufiuc

    2013-01-01

    Full Text Available The present work reports a rapid, simple and efficient one-step synthesis and detailed characterisation of stable aqueous colloids of gold nanoparticles (AuNPs coated with unmodified poly(ethyleneglycol (PEG molecules of different molecular weights and surface charges. By mixing and heating aqueous solutions of PEG with variable molecular chain and gold(III chloride hydrate (HAuCl4 in the presence of NaOH, we have successfully produced uniform colloidal 5 nm PEG coated AuNPs of spherical shape with tunable surface charge and an average diameter of 30 nm within a few minutes. It has been found out that PEGylated AuNPs provide optical enhancement of the characteristic vibrational bands of PEG molecules attached to the gold surface when they are excited with both visible (532 nm and NIR (785 nm laser lines. The surface enhanced Raman scattering (SERS signal does not depend on the length of the PEG molecular chain enveloping the AuNPs, and the stability of the colloid is not affected by the addition of concentrated salt solution (0.1 M NaCl, thus suggesting their potential use for in vitro and in vivo applications. Moreover, by gradually changing the chain length of the biopolymer, we were able to control nanoparticles’ surface charge from −28 to −2 mV, without any modification of the Raman enhancement properties and of the colloidal stability.

  4. One-step purification of soluble recombinant human 6-phosphogluconate dehydrogenase from Escherichia coli.

    Science.gov (United States)

    Chan, Barden; Sukhatme, Vikas P

    2013-11-01

    6-Phosphogluconate dehydrogenase (6PGD), the third enzyme in the pentose phosphate pathway, was recently identified as a novel target in human lung cancer. In this report, we present an expression and purification scheme of recombinant human 6PGD from Escherichia coli. Using a DE3 derivative strain expressing tRNAs for seven rare codons in E. coli called Rosetta2 (DE3), a large quantity of soluble human 6PGD can be expressed with an N-terminal histidine tag and purified by a one-step purification procedure to near homogeneity without denaturants or refolding. Three to seven milligrams of purified protein could be obtained from 100 ml of culture. This recombinant human 6PGD follows classic Michaelis-Menton saturation kinetics with respect to both substrates NADP(+) and 6-phosphogluconate. The respective k(cat) and K(m) were comparable to those of 6PGDs purified from mammalian tissues. Using this purified 6PGD enzyme, we devised an endpoint colorimetric assay suitable for high-throughput screening for human 6PGD inhibitors.

  5. Improved adhesion of superhydrophobic layer on metal surfaces via one step spraying method

    Directory of Open Access Journals (Sweden)

    Wael I. El Dessouky

    2017-03-01

    Full Text Available Superhydrophobic metal substrates have been fabricated by a simple spraying method. The processes of decreasing surface free energy and increasing surface roughness have been accomplished in one step via the addition of functionalized silica (silica nano particles with octyltriethoxysilane to adhesive polymer. The method is simple, cost-effective and can be applied on the large industrial scale. Scanning electron microscopy (SEM was used for surface morphology analysis, showing the roughness produced by surface treatment. The wettability of the micro-nano silica film varied from hydrophilicity (water contact angle 88° to superhydrophobicity (water contact angle 156.9°, while sliding contact angles dramatically decreased (<5° by adding Functionalized silica and/or adhesive polymer. Roughness increased with silica increment which improves the wettability. The coatings were electrochemically characterized by electrochemical impedance spectroscopy (EIS and Tafel polarization curves; it was found that both systems had good performance against corrosion in 3.5% sodium chloride solution. Furthermore, the stability of the coated layer on copper substrate was investigated.

  6. One-step electrodeposition process to fabricate corrosion-resistant superhydrophobic surface on magnesium alloy.

    Science.gov (United States)

    Liu, Qin; Chen, Dexin; Kang, Zhixin

    2015-01-28

    A simple, one-step method has been developed to construct a superhydrophobic surface by electrodepositing Mg-Mn-Ce magnesium plate in an ethanol solution containing cerium nitrate hexahydrate and myristic acid. Scanning electron microscopy, energy-dispersive X-ray spectroscopy, X-ray photoelectron spectroscopy, and Fourier transform infrared spectroscopy were employed to characterize the surfaces. The shortest electrodeposition time to obtain a superhydrophobic surface was about 1 min, and the as-prepared superhydrophobic surfaces had a maximum contact angle of 159.8° and a sliding angle of less than 2°. Potentiodynamic polarization and electrochemical impedance spectroscopy measurements demonstrated that the superhydrophobic surface greatly improved the corrosion properties of magnesium alloy in 3.5 wt % aqueous solutions of NaCl, Na2SO4, NaClO3, and NaNO3. Besides, the chemical stability and mechanical durability of the as-prepared superhydrophobic surface were also examined. The presented method is rapid, low-cost, and environmentally friendly and thus should be of significant value for the industrial fabrication of anticorrosive superhydrophobic surfaces and should have a promising future in expanding the applications of magnesium alloys.

  7. Nearly Perfect Durable Superhydrophobic Surfaces Fabricated by a Simple One-Step Plasma Treatment.

    Science.gov (United States)

    Ryu, Jeongeun; Kim, Kiwoong; Park, JooYoung; Hwang, Bae Geun; Ko, YoungChul; Kim, HyunJoo; Han, JeongSu; Seo, EungRyeol; Park, YongJong; Lee, Sang Joon

    2017-05-16

    Fabrication of superhydrophobic surfaces is an area of great interest because it can be applicable to various engineering fields. A simple, safe and inexpensive fabrication process is required to fabricate applicable superhydrophobic surfaces. In this study, we developed a facile fabrication method of nearly perfect superhydrophobic surfaces through plasma treatment with argon and oxygen gases. A polytetrafluoroethylene (PTFE) sheet was selected as a substrate material. We optimized the fabrication parameters to produce superhydrophobic surfaces of superior performance using the Taguchi method. The contact angle of the pristine PTFE surface is approximately 111.0° ± 2.4°, with a sliding angle of 12.3° ± 6.4°. After the plasma treatment, nano-sized spherical tips, which looked like crown-structures, were created. This PTFE sheet exhibits the maximum contact angle of 178.9°, with a sliding angle less than 1°. As a result, this superhydrophobic surface requires a small external force to detach water droplets dripped on the surface. The contact angle of the fabricated superhydrophobic surface is almost retained, even after performing an air-aging test for 80 days and a droplet impacting test for 6 h. This fabrication method can provide superb superhydrophobic surface using simple one-step plasma etching.

  8. One-step green synthesis of bimetallic Fe/Pd nanoparticles used to degrade Orange II.

    Science.gov (United States)

    Luo, Fang; Yang, Die; Chen, Zuliang; Megharaj, Mallavarapu; Naidu, Ravendra

    2016-02-13

    To reduce cost and enhance reactivity, bimetallic Fe/Pd nanoparticles (NPs) were firstly synthesized using grape leaf aqueous extract to remove Orange II. Green synthesized bimetallic Fe/Pd NPs (98.0%) demonstrated a far higher ability to remove Orange II in 12h compared to Fe NPs (16.0%). Meanwhile, all precursors, e.g., grape leaf extract, Fe(2+) and Pd(2+), had no obvious effect on removing Orange II since less than 2.0% was removed. Kinetics study revealed that the removal rate fitted well to the pseudo-first-order reduction and pseudo-second-order adsorption model, meaning that removing Orange II via Fe/Pd NPs involved both adsorption and catalytic reduction. The remarkable stability of Fe/Pd NPs showed the potential application for removing azo dyes. Furthermore, Scanning Electron Microscopy (SEM) and Fourier Transform Infrared Spectroscopy (FTIR) confirmed the changes in Fe/Pd NPs before and after reaction with Orange II. High Performance Liquid Chromatography-Mass Spectrum (HPLC-MS) identified the degraded products in the removal of Orange II, and finally a removal mechanism was proposed. This one-step strategy using grape leaf aqueous extract to synthesize Fe/Pd NPs is simple, cost-effective and environmentally benign, making possible the large-scale production of Fe/Pd NPs for field remediation. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. One-step synthesis and microstructure of CuO-SDC composites

    Energy Technology Data Exchange (ETDEWEB)

    Firmino, H.C.T.; Araujo, A.J.M.; Dutra, R.P.S.; Macedo, D.A., E-mail: hellentorrano@hotmail.com, E-mail: allanjp1993@hotmail.com, E-mail: ricardopsd@gmail.com, E-mail: damaced@gmail.com [Universidade Federal da Paraiba (UFPB), Joao Pessoa, PB (Brazil); Nascimento, R.M., E-mail: rmaribondo@ufrnet.br [Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN (Brazil); Rajesh, S., E-mail: rajeshayr@gmail.com [University of Aveiro (Portugal)

    2017-01-15

    An in situ one step synthesis route based on the polymeric precursor method was used to produce dual phase CuO-samaria doped ceria (SDC) nanocomposite powders. This chemical route allowed to obtain composite powders with reduced particle size and uniform distribution of Cu, Ce and Sm elements. The particulate material was characterized by powder X-ray diffraction (XRD) combined with Rietveld refinement. CuO-SDC sintered in air between 950 to 1050 °C and subsequently reduced to Cu-SDC cermets were further characterized by XRD and scanning electron microscopy. The open porosity was measured using the Archimedes' principle. Suitable microstructures for both charge transfer and mass transport processes (30 to 45% porosity) were attained in Cu-SDC cermets previously fired at 1000 to 1050 °C. Overall results indicated that CuO-SDC composites and Cu-SDC cermets with potential application as anodes for solid oxide fuel cells (SOFCs) can be obtained by microstructural design. An anode supported half-cell was prepared by co-pressing and co-firing gadolinia doped ceria (CGO) and the herein synthesized CuO-SDC nanocomposite powder. (author)

  10. Fluorinated alkyne-derived monolayers on oxide-free silicon nanowires via one-step hydrosilylation

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen Minh, Quyen [Laboratory of Organic Chemistry, Wageningen University, Stippeneng 4, 6708 WE Wageningen (Netherlands); Nanosens, IJsselkade 7, 7201 HB Zutphen (Netherlands); Pujari, Sidharam P. [Laboratory of Organic Chemistry, Wageningen University, Stippeneng 4, 6708 WE Wageningen (Netherlands); Wang, Bin [The Department of Chemical Engineering and Russell Berrie Nanotechnology Institute, Technion – Israel Institute of Technology, Haifa 3200003 (Israel); Wang, Zhanhua [Laboratory of Organic Chemistry, Wageningen University, Stippeneng 4, 6708 WE Wageningen (Netherlands); Haick, Hossam [The Department of Chemical Engineering and Russell Berrie Nanotechnology Institute, Technion – Israel Institute of Technology, Haifa 3200003 (Israel); Zuilhof, Han [Laboratory of Organic Chemistry, Wageningen University, Stippeneng 4, 6708 WE Wageningen (Netherlands); Rijn, Cees J.M. van, E-mail: cees.vanrijn@wur.nl [Laboratory of Organic Chemistry, Wageningen University, Stippeneng 4, 6708 WE Wageningen (Netherlands)

    2016-11-30

    Highlights: • Oxide-free H-terminated silicon nanowires undergo efficient surface modification by reaction with fluorinated 1-alkynes (HC≡C−(CH{sub 2}){sub 6}C{sub 8}H{sub 17−x}F{sub x}; x = 0–17). • These surface-modified Si NWs are chemically stable under range of conditions (including acid, base). • The surface coating yields efficient electrical passivation as demonstrated by a near-zero electrochemical activity of the surface. - Abstract: Passivation of oxide-free silicon nanowires (Si NWs) by the formation of high-quality fluorinated 1-hexadecyne-derived monolayers with varying fluorine content has been investigated. Alkyl chain monolayers (C{sub 16}H{sub 30−x}F{sub x}) with a varying number of fluorine substituents (x = 0, 1, 3, 9, 17) were attached onto hydrogen-terminated silicon (Si−H) surfaces with an effective one-step hydrosilylation. This surface chemistry gives well-defined monolayers on nanowires that have a cylindrical core–shell structure, as characterized by X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FT-IR) and static contact angle (SCA) analysis. The monolayers were stable under acidic and basic conditions, as well as under extreme conditions (such as UV exposure), and provide excellent surface passivation, which opens up applications in the fields of field effect transistors, optoelectronics and especially for disease diagnosis.

  11. InGaN directional coupler made with a one-step etching technique

    Science.gov (United States)

    Gao, Xumin; Yuan, Jialei; Yang, Yongchao; Zhang, Shuai; Shi, Zheng; Li, Xin; Wang, Yongjin

    2017-06-01

    We propose, fabricate and characterize an on-chip integration of light source, InGaN waveguide, directional coupler and photodiode, in which AlGaN layers are used as top and bottom optical claddings to form an InGaN waveguide for guiding the in-plane emitted light from the InGaN/GaN multiple-quantum-well light-emitting diode (MQW-LED). The difference in etch rate caused by different exposure windows leads to an etching depth discrepancy using the one-step etching technique, which forms the InGaN directional coupler with the overlapped underlying slab. Light propagation results directly confirm effective light coupling in the InGaN directional coupler, which is achieved through high-order guided modes. The InGaN waveguide couples the modulated light from the InGaN/GaN MQW-LED and transfers part of light to the coupled waveguide via the InGaN directional coupler. The in-plane InGaN/GaN MQW-photodiode absorbs the guided light by the coupled InGaN waveguide and induces the photocurrent. The on-chip InGaN photonic integration experimentally demonstrates an in-plane light communication with a data transmission of 50 Mbps.

  12. A one pot, one step, precision cloning method with high throughput capability.

    Directory of Open Access Journals (Sweden)

    Carola Engler

    Full Text Available Current cloning technologies based on site-specific recombination are efficient, simple to use, and flexible, but have the drawback of leaving recombination site sequences in the final construct, adding an extra 8 to 13 amino acids to the expressed protein. We have devised a simple and rapid subcloning strategy to transfer any DNA fragment of interest from an entry clone into an expression vector, without this shortcoming. The strategy is based on the use of type IIs restriction enzymes, which cut outside of their recognition sequence. With proper design of the cleavage sites, two fragments cut by type IIs restriction enzymes can be ligated into a product lacking the original restriction site. Based on this property, a cloning strategy called 'Golden Gate' cloning was devised that allows to obtain in one tube and one step close to one hundred percent correct recombinant plasmids after just a 5 minute restriction-ligation. This method is therefore as efficient as currently used recombination-based cloning technologies but yields recombinant plasmids that do not contain unwanted sequences in the final construct, thus providing precision for this fundamental process of genetic manipulation.

  13. One-step synthesis and microstructure of CuO-SDC composites

    Directory of Open Access Journals (Sweden)

    H. C. T. Firmino

    Full Text Available Abstract An in situ one step synthesis route based on the polymeric precursor method was used to produce dual phase CuO-samaria doped ceria (SDC nanocomposite powders. This chemical route allowed to obtain composite powders with reduced particle size and uniform distribution of Cu, Ce and Sm elements. The particulate material was characterized by powder X-ray diffraction (XRD combined with Rietveld refinement. CuO-SDC sintered in air between 950 to 1050 °C and subsequently reduced to Cu-SDC cermets were further characterized by XRD and scanning electron microscopy. The open porosity was measured using the Archimedes’ principle. Suitable microstructures for both charge transfer and mass transport processes (30 to 45% porosity were attained in Cu-SDC cermets previously fired at 1000 to 1050 °C. Overall results indicated that CuO-SDC composites and Cu-SDC cermets with potential application as anodes for solid oxide fuel cells (SOFCs can be obtained by microstructural design. An anode supported half-cell was prepared by co-pressing and co-firing gadolinia doped ceria (CGO and the herein synthesized CuO-SDC nanocomposite powder.

  14. One-step electrochemical deposition of Schiff base cobalt complex as effective water oxidation catalyst

    Science.gov (United States)

    Huang, Binbin; Wang, Yan; Zhan, Shuzhong; Ye, Jianshan

    2017-02-01

    Schiff base metal complexes have been applied in many fields, especially, a potential homogeneous catalyst for water splitting. However, the high overpotential, time consumed synthesis process and complicated working condition largely limit their application. In the present work, a one-step approach to fabricate Schiff base cobalt complex modified electrode is developed. Microrod clusters (MRC) and rough spherical particles (RSP) can be obtained on the ITO electrode through different electrochemical deposition condition. Both of the MRC and RSP present favorable activity for oxygen evolution reaction (OER) compared to the commercial Co3O4, taking an overpotential of 650 mV and 450 mV to drive appreciable catalytic current respectively. The highly active and stable RSP shows a Tafel plot of 84 mV dec-1 and negligible decrease of the current density for 12 h bulk electrolysis. The synthesis strategy of effective and stable catalyst in this work provide a simple method to fabricate heterogeneous OER catalyst with Schiff base metal complex.

  15. ONE-STEP PROCESS TO MAKE ELECTRICALLY CONDUCTIVE THERMOPLASTIC VULCANIZATES FILLED WITH MWCNTs

    Institute of Scientific and Technical Information of China (English)

    Hao-sheng Wang; Xiao-hong Zhang; Yi-lei Zhu; Zhi-hai Song; Jin-liang Qiao

    2012-01-01

    Electrically conductive thermoplastic vulcanizates (TPVs) filled by multi-walled carbon nanotubes (MWCNTs)are prepared by a simple one-step melt mixing process,based on linear low density polyethylene (LLDPE) and ultrafine full-vulcanized rubber particles (UFRP).An ideal morphology with controlled localization of MWCNTs in continuous LLDPE matrix and appropriate size of finely-dispersed UFRP can be achieved at the same time.The controlled localization of MWCNTs in the continuous phase facilitates the formation of conductive pathway,and thus the volume resistivity of the as-prepared LLDPE/UFRP/MWCNTs thermoplastic vulcanizates is significantly decreased.The results show that both the blend ratio of LLDPE/UFRP and the loading of MWCNTs have remarkable effect on the volume resistivity.Significantly,the electrically conductive TPVs exhibit good mechanical properties duo to the fine dispersion of UFRP in LLDPE.The added MWCNTs are capable of imparting reinforcement effects to thermoplastic vulcanizates with just a slight loss of stretchability and elasticity.

  16. Enhancement of valve metal osteoconductivity by one-step hydrothermal treatment

    Energy Technology Data Exchange (ETDEWEB)

    Zuldesmi, Mansjur, E-mail: mzuldesmi@yahoo.com [Department of Materials Science and Engineering, Graduate School of Engineering, Nagoya University, Nagoya (Japan); Department of Mechanical Engineering, Manado State University (UNIMA) (Indonesia); Waki, Atsushi [Department of Materials Science and Engineering, Graduate School of Engineering, Nagoya University, Nagoya (Japan); Kuroda, Kensuke; Okido, Masazumi [EcoTopia Science Institute, Nagoya University, Nagoya (Japan)

    2014-09-01

    In this study, we produced super-hydrophilic surfaces of valve metals (Ti, Nb, Ta and Zr) by one-step hydrothermal treatment. Their surface characteristics and osteoconductivity using an in vivo test were then assessed. These data were compared with that of as-polished, as-anodized and both anodized + hydrothermally treated samples. Changes in surface chemistry, surface morphology and structure were investigated by X-ray photoelectron spectroscopy, scanning electron microscopy, and X-ray diffractometry. The results revealed that the water contact angles of valve metals were decreased by hydrothermal treatment and continued to reduce dramatically until lower than 10° after being immersed in phosphate buffered solution. By producing super-hydrophilic surfaces, the osteoconductivity of these hydrothermally treated valve metals was enhanced by up to 55%. - Highlights: • Hydrothermal treatment in distilled water was applied to valve metals. • Surface characteristics and osteoconductivity by in vivo test were evaluated. • Water contact angles of valve metals were decreased by hydrothermal treatment. • Osteoconductivity of valve metals improved notably by hydrothermal treatment.

  17. One-step preparation of hydrogenated ZrO2 microspheres by cathode plasma electrolysis

    Science.gov (United States)

    Liu, Chenxu; Xiang, Qingyun; Yang, Mu; Wang, Shengdian; Wang, Linxiu; Zhang, Jin; He, Yedong

    2017-07-01

    Hydrogenated ZrO2 microspheres were directly prepared by cathode plasma electrolysis (CPE) in an aqueous solution of Zr(NO3)4•5H2O. Owing to the energy of plasma and the cathodic hydrogen evolution reactions, the CPE method combined the preparation of ZrO2 ceramic and the hydrogen treatment into only one step. The results showed regular microspheres consisting of tetragonal-ZrO2 and monoclinic-ZrO2 with 1-10 µm in diameter were formed at relatively high concentration of Zr(NO3)3•5H2O. These ZrO2 microspheres contained about 52.54 µg g-1 hydrogen which caused a narrow band gap (3.10 eV). Thus, the microspheres showed good photocatalytic activity under simulated sunlight, and the degradation of RhB dye reached nearly 58% for 3 h of irradiation, much better than the ZrO2 microspheres after dehydrogenation treatment.

  18. One-step sol-gel imprint lithography for guided-mode resonance structures.

    Science.gov (United States)

    Huang, Yin; Liu, Longju; Johnson, Michael; C Hillier, Andrew; Lu, Meng

    2016-03-01

    Guided-mode resonance (GMR) structures consisting of sub-wavelength periodic gratings are capable of producing narrow-linewidth optical resonances. This paper describes a sol-gel-based imprint lithography method for the fabrication of submicron 1D and 2D GMR structures. This method utilizes a patterned polydimethylsiloxane (PDMS) mold to fabricate the grating coupler and waveguide for a GMR device using a sol-gel thin film in a single step. An organic-inorganic hybrid sol-gel film was selected as the imprint material because of its relatively high refractive index. The optical responses of several sol-gel GMR devices were characterized, and the experimental results were in good agreement with the results of electromagnetic simulations. The influence of processing parameters was investigated in order to determine how finely the spectral response and resonant wavelength of the GMR devices could be tuned. As an example potential application, refractometric sensing experiments were performed using a 1D sol-gel device. The results demonstrated a refractive index sensitivity of 50 nm/refractive index unit. This one-step fabrication process offers a simple, rapid, and low-cost means of fabricating GMR structures. We anticipate that this method can be valuable in the development of various GMR-based devices as it can readily enable the fabrication of complex shapes and allow the doping of optically active materials into sol-gel thin film.

  19. [One-step PCR sequencing]. Final report, July 1, 1994--August 31, 1997

    Energy Technology Data Exchange (ETDEWEB)

    Shaw, B.R.

    1997-12-31

    The author explored the use of a novel class of boronated nucleic acids, the boranophosphates, as an alternative, but complementary method to dideoxysequencing. Boranophosphates can be used to directly amplify and sequence single- or double-stranded DNA. Fragments are derived not from truncations during polymerase synthesis, but from insertion and digestion back to a boronated marker or delimiter that was incorporated during exponential amplification. The method, which the author calls Boronated One-Step PCR Sequencing, is unique in that it employs a new class of {alpha}-P-boronated 2{prime}-deoxynucleoside 5{prime}-triphosphates first synthesized in the laboratory. These boronated triphosphates exhibit useful properties: (a) they are heat stable, (b) they can be base-specifically incorporated into DNA during the polymerase chain reaction, and (c) once incorporated, the boranophosphate nucleotide (marker) blocks the action of exonuclease. Thus, the positions of the stably-incorporated boronated markers can be revealed by a simple exonuclease digestion, producing a series of fragments--each of which is terminated base-specifically at the boronated markers--and thereby defining the sequence of the PCR product.

  20. One-step selection of Vaccinia virus-binding DNA aptamers by MonoLEX

    Directory of Open Access Journals (Sweden)

    Stöcklein Walter

    2007-08-01

    Full Text Available Abstract Background As a new class of therapeutic and diagnostic reagents, more than fifteen years ago RNA and DNA aptamers were identified as binding molecules to numerous small compounds, proteins and rarely even to complete pathogen particles. Most aptamers were isolated from complex libraries of synthetic nucleic acids by a process termed SELEX based on several selection and amplification steps. Here we report the application of a new one-step selection method (MonoLEX to acquire high-affinity DNA aptamers binding Vaccinia virus used as a model organism for complex target structures. Results The selection against complete Vaccinia virus particles resulted in a 64-base DNA aptamer specifically binding to orthopoxviruses as validated by dot blot analysis, Surface Plasmon Resonance, Fluorescence Correlation Spectroscopy and real-time PCR, following an aptamer blotting assay. The same oligonucleotide showed the ability to inhibit in vitro infection of Vaccinia virus and other orthopoxviruses in a concentration-dependent manner. Conclusion The MonoLEX method is a straightforward procedure as demonstrated here for the identification of a high-affinity DNA aptamer binding Vaccinia virus. MonoLEX comprises a single affinity chromatography step, followed by subsequent physical segmentation of the affinity resin and a single final PCR amplification step of bound aptamers. Therefore, this procedure improves the selection of high affinity aptamers by reducing the competition between aptamers of different affinities during the PCR step, indicating an advantage for the single-round MonoLEX method.

  1. Biocompatible gold nanorods: one-step surface functionalization, highly colloidal stability, and low cytotoxicity.

    Science.gov (United States)

    Liu, Kang; Zheng, Yuanhui; Lu, Xun; Thai, Thibaut; Lee, Nanju Alice; Bach, Udo; Gooding, J Justin

    2015-05-05

    The conjugation of gold nanorods (AuNRs) with polyethylene glycol (PEG) is one of the most effective ways to reduce their cytotoxicity arising from the cetyltrimethylammonium bromide (CTAB) and silver ions used in their synthesis. However, typical PEGylation occurs only at the tips of the AuNRs, producing partially modified AuNRs. To address this issue, we have developed a novel, facile, one-step surface functionalization method that involves the use of Tween 20 to stabilize AuNRs, bis(p-sulfonatophenyl)phenylphosphine (BSPP) to activate the AuNR surface for the subsequent PEGylation, and NaCl to etch silver from the AuNRs. This method allows for the complete removal of the surface-bound CTAB and the most active surface silver from the AuNRs. The produced AuNRs showed far lower toxicity than other methods to PEGylate AuNRs, with no apparent toxicity when their concentration is lower than 5 μg/mL. Even at a high concentration of 80 μg/mL, their cell viability is still four times higher than that of the tip-modified AuNRs.

  2. Rapid synthesis of gold nanorods using a one-step photochemical strategy.

    Science.gov (United States)

    Ahmed, Marya; Narain, Ravin

    2010-12-07

    Rapid synthesis of gold nanorods of controlled dimensions is one of the desired aspects of nanotechnology as a result of the potential of these nanomaterials for biomedical applications. The synthesis of gold nanorods has been achieved using a photoinitiator as an instant source of ketyl radicals, which allows the synthesis of gold nanorods in minutes. This is the first report providing a one-step synthesis of nanorods of controlled dimensions in 20-30 min using photoinitiator I-2959 as a source of ketyl radicals. Furthermore, the role of UV intensity, the concentration of silver ions, and the presence of cosolvents and a cosurfactant have been studied in detail in an effort to produce nanorods with controlled dimensions in higher yields. The role of acetone in nanorod synthesis has been explored in detail, and it has been demonstrated that, for the photochemical synthesis of nanorods using a photoinitiator, acetone is not a critical component and can be replaced by other water-miscible solvents, thus the successful synthesis of nanorods in tetrahydrofuran (THF) has been demonstrated. It has also been found that a cosurfactant and an organic solvent are not required for the synthesis of nanorods; however, their presence is found to improve the monodispersity of nanorod samples, in addition to providing a higher yield.

  3. One-step Solution Processing of Ag, Au and Pd@MXene Hybrids for SERS

    Science.gov (United States)

    Satheeshkumar, Elumalai; Makaryan, Taron; Melikyan, Armen; Minassian, Hayk; Gogotsi, Yury; Yoshimura, Masahiro

    2016-08-01

    We report on one-step hybridization of silver, gold and palladium nanoparticles from solution onto exfoliated two-dimensional (2D) Ti3C2 titanium carbide (MXene) nanosheets. The produced hybrid materials can be used as substrates for surface-enhanced Raman spectroscopy (SERS). An approximate analytical approach is also developed for the calculation of the surface plasmon resonance (SPR) frequency of nanoparticles immersed in a medium, near the interface of two dielectric media with different dielectric constants. We obtained a good match with the experimental data for SPR wavelengths, 440 nm and 558 nm, respectively for silver and gold nanoparticles. In the case of palladium, our calculated SPR wavelength for the planar geometry was 160 nm, demonstrating that non-spherical palladium nanoparticles coupled with 2D MXene yield a broad, significanlty red-shifted SPR band with a peak at 230 nm. We propose a possible mechanism of the plasmonic hybridization of nanoparticles with MXene. The as-prepared noble metal nanoparticles on MXene show a highly sensitive SERS detection of methylene blue (MB) with calculated enhancement factors on the order of 105. These findings open a pathway for extending visible-range SERS applications of novel 2D hybrid materials in sensors, catalysis, and biomedical applications.

  4. Synthesis and Characteristics of Mesoporous Silica Aerogels with One-step Solvent Exchange/Surface Modification

    Institute of Scientific and Technical Information of China (English)

    WANG Lijiu; ZHAO Shanyu

    2009-01-01

    The synthesis procedures and physical properties of the ambient dried hydrophobic silica aerogels by using different contents of ethanol(EtOH)/trimethylchlorosilane(TMCS)/n-Hexane as surface modification agent were investigated.One-step solvent exchange and surface modification were simultaneously progressed by immersing silica hydrogels in EtOH/TMCS/n-Hexane solution.It is found that microstructures as well as properties of silica aerogels like porosity,specific density and specific surface area are affected by the contents of surface modification agent in the sol from the re-sults of SEM,TEM morphology,FT-IR chemical structure,BET surface area and BJH pore size analyses.The volume of TMCS is of 10%and 20%of hydrogels,and the final product is hydrophilic xerogels.When TMCS's percent(v/v)is elevated to 75%-100%,hydrophobic silica aerogels with good performance are synthesized,the porosities of aerogels are in the range of 93.5%-95.8%and the av-erage pore size diameter is less than 20 nm.

  5. Anisotropic wetting of copper alloys induced by one-step laser micro-patterning

    Energy Technology Data Exchange (ETDEWEB)

    Hans, M., E-mail: michael.hans@mx.uni-saarland.de [Chair of Functional Materials, Faculty of Natural Sciences and Technology, Saarland University, 66123 Saarbruecken (Germany); Mueller, F.; Grandthyll, S.; Huefner, S. [Experimental Physics, Faculty of Natural Sciences and Technology, Saarland University, 66123 Saarbruecken (Germany); Muecklich, F. [Chair of Functional Materials, Faculty of Natural Sciences and Technology, Saarland University, 66123 Saarbruecken (Germany)

    2012-12-15

    Highlights: Black-Right-Pointing-Pointer One-step, contactless micro-patterning of copper alloys has been achieved. Black-Right-Pointing-Pointer Anisotropic wetting properties are tailored by line-like structures. Black-Right-Pointing-Pointer Both topographical and chemical patterns contribute to the phenomenon. Black-Right-Pointing-Pointer The topographic shape and homogeneity are found to be governing factors. - Abstract: Copper alloys (CuSn8, CuZn23Al3Co) have been micro-patterned with line-like geometries by Laser Interference Surface Structuring (LISS). In the presented study two high power pulsed laser beams are recombined to create unique, line-like intensity distributions with a chosen, constant periodicity of 10 {mu}m at varying laser fluencies. Anisotropic wetting properties on these surfaces have been confirmed by drop shape analysis and static contact angle measurements, which were conducted parallel and perpendicular to the structures revealing up to 25% difference in contact angle. The topography and chemistry of the tailored line structures have been characterized and analyzed by white light interferometry, spatial frequency distribution, AFM and X-ray photoelectron spectroscopy. The topographic shape and homogeneity are considered as key parameters for anisotropic wetting design, although it is concluded that both, the geometry as well as the locally varying chemical composition of the surface structures contribute to the phenomenon. Parallel capillarity effects and perpendicular contact line pinning are found to be the governing mechanisms.

  6. Epoxy Resins Toughened with Surface Modified Epoxidized Natural Rubber Fibers by One-Step Electrospinning.

    Science.gov (United States)

    Kim, Joo Ran; Kim, Jung J

    2017-04-27

    Epoxidized natural rubber fibers (ERFs) are developed through one-step electrospinning and directly deposited into epoxy resins without collecting and distributing of fibers. The shape of ERFs shows rough surface due to different evaporation rate of solvent mixture consisting of chloroform and dichloromethane and the average diameter of ERFs is 6.2 µm. The increase of ERFs loading from 0 to 20 wt % into the epoxy resin increases the fracture strain significantly from 1.2% to 13% and toughness from 0.3 MPa to 1.9 MPa by a factor of 7. However, the tensile strength and Young's modulus decrease about 34% from 58 MPa to 34 MPa and from 1.4 GPa to 0.9 GPa, respectively. Due to the crosslinking reactions between oxirane groups of ERFs and amine groups in the resin, surface roughness and the high aspect ratio of ERFs, ERFs result in more effective toughening effect with the minimum loss of tensile properties in epoxy resins.

  7. Optimality analysis of one-step OOSM filtering algorithms in target tracking

    Institute of Scientific and Technical Information of China (English)

    ZHOU WenHui; LI Lin; CHEN GuoHai; YU AnXi

    2007-01-01

    In centralized multisensor tracking systems, there are out-of-sequence measurements (OOSMs) frequently arising due to different time delays in communication links and varying pre-processing times at the sensor. Such OOSM arrival can induce the "negative-time measurement update" problem, which is quite common in real multisensor tracking systems. The A1 optimal update algorithm with OOSM is presented by Bar-Shalom for one-step case. However, this paper proves that the optimality of A1 algorithm is lost in direct discrete-time model (DDM) of the process noise, it holds true only in discretized continuous-time model (DCM). One better OOSM filtering algorithm for DDM case is presented. Also, another new optimal OOSM filtering algorithm, which is independent of the discrete time model of the process noise, is presented here. The performance of the two new algorithms is compared with that of A1 algorithm by Monte Carlo simulations. The effectiveness and correctness of the two proposed algorithms are validated by analysis and simulation results.

  8. Optimisation of one-step desolvation and scale-up of gelatine nanoparticle production.

    Science.gov (United States)

    Geh, Katharina J; Hubert, Madlen; Winter, Gerhard

    2016-11-01

    Gelatine nanoparticles (GNPs) are biodegradable and biocompatible drug delivery systems with excellent clinical performances. A two-step desolvation is commonly used for their preparation, although this methodology has several shortcomings: lack of reproducibility, small scales and low yields. A straightforward and more consistent GNP preparation approach is presented here focusing on the development of a one-step desolvation with the use of a commercially available gelatine type. Controlled stirring conditions and ultrafiltration are used to achieve large-scale production of nanoparticles of up to 2.6 g per batch. Particle size distributions are conserved and comparable to those determined for two-step desolvation on small scale. Additionally, a range of cross-linking agents is examined for their effectiveness in stabilising GNPs as an alternative to glutaraldehyde. Glyceraldehyde demonstrated outstanding properties, which led to high colloidal stability. This approach optimises the manufacturing process and the scale-up of the production capacity, providing a clear potential for future applications.

  9. Parallel multifunctionalization of nanoparticles: a one-step modular approach for in vivo imaging.

    Science.gov (United States)

    Groult, Hugo; Ruiz-Cabello, Jesús; Pellico, Juan; Lechuga-Vieco, Ana V; Bhavesh, Riju; Zamai, Moreno; Almarza, Elena; Martín-Padura, Inés; Cantelar, Eugenio; Martínez-Alcázar, María P; Herranz, Fernando

    2015-01-21

    Multifunctional nanoparticles are usually produced by sequential synthesis, with long multistep protocols. Our study reports a generic modular strategy for the parallel one-step multifunctionalization of different hydrophobic nanoparticles. The method was designed and developed by taking advantage of the natural noncovalent interactions between the fatty acid binding sites of the bovine serum albumin (BSA) and the aliphatic surfactants on different inorganic nanomaterials. As a general example of the approach, three different nanoparticles-iron oxide, upconverting nanophosphors, and gold nanospheres-were nanoemulsified in water with BSA. To support specific applications, multifunctional capability was incorporated with a variety of previously modified BSA modules. These modules include different conjugated groups, such as chelating agents for (68)Ga or (89)Zr and ligand molecules for enhanced in vivo targeting. A large library of 13 multimodal contrast agents was developed with this convergent strategy. This platform allows a highly versatile and easy tailoring option for efficient incorporation of functional groups. Finally, as demonstration of this versatility, a bimodal (PET/MRI) probe including a maleimide-conjugated BSA was selectively synthesized with an RGD peptide for in vivo imaging detection of tumor angiogenesis.

  10. Cryopreservation of Pistacia spp. seeds by dehydration and one-step freezing.

    Science.gov (United States)

    Ozden-Tokatli, Y; Ozudogru, E A; Gumusel, F; Lambardi, M

    2007-01-01

    Cryopreservation protocols by dehydration and one-step freezing were developed for seeds from three Pistacia species, i.e., P. vera, P. terebinthus and P. lentiscus, which were characterised by different initial germination percentages (100%, 17% and 81%, respectively). In P. vera, a maximum of 90% germination was obtained following 8 hours drying in silica gel (corresponding to 11.7% moisture content on a FW basis) and direct immersion in LN. In P. terebinthus and P. lentiscus, shorter periods of dehydration (1 hour and 15 min, respectively) were sufficient to reduce their moisture content to about 20%, which resulted in peak seed germination percentages from cryostorage of 16% and 47%, respectively. Following cryopreservation, the seeds germinated better on semi-solid MS medium, than on cotton wool wetted with dH(2)O or liquid MS medium. Finally, in P. vera and P. lentiscus, high and significant correlation coefficients were obtained between the TTC viability test and seed germinability after recovery from LN, provided that seeds which were considered positive in the test showed completely or partially red embryonic axes coupled to completely red cotyledons.

  11. Demosaicking by alternating projections: theory and fast one-step implementation.

    Science.gov (United States)

    Lu, Yue M; Karzand, Mina; Vetterli, Martin

    2010-08-01

    Color image demosaicking is a key process in the digital imaging pipeline. In this paper, we study a well-known and influential demosaicking algorithm based upon alternating projections (AP), proposed by Gunturk, Altunbasak and Mersereau in 2002. Since its publication, the AP algorithm has been widely cited and compared against in a series of more recent papers in the demosaicking literature. Despite good performances, a limitation of the AP algorithm is its high computational complexity. We provide three main contributions in this paper. First, we present a rigorous analysis of the convergence property of the AP demosaicking algorithm, showing that it is a contraction mapping, with a unique fixed point. Second, we show that this fixed point is in fact the solution to a constrained quadratic minimization problem, thus, establishing the optimality of the AP algorithm. Finally, using the tool of polyphase representation, we show how to obtain the results of the AP algorithm in a single step, implemented as linear filtering in the polyphase domain. Replacing the original iterative procedure by the proposed one-step solution leads to substantial computational savings, by about an order of magnitude in our experiments.

  12. One-step bone marrow-derived cell transplantation in talarosteochondral lesions: mid-term results.

    Science.gov (United States)

    Buda, Roberto; Vannini, Francesca; Cavallo, Marco; Baldassarri, Matteo; Natali, Simone; Castagnini, Francesco; Giannini, Sandro

    2013-01-01

    to verify the capability of scaffold-supported bone marrow-derived cells to be used in the repair of osteochondral lesions of the talus. using a device to concentrate bone marrow-derived cells, a scaffold (collagen powder or hyaluronic acid membrane) for cell support and platelet gel, a one-step arthroscopic technique was developed for cartilage repair. In a prospective clinical study, we investigated the ability of this technique to repair talar osteochondral lesions in 64 patients. The mean follow-up was 53 months. Clinical results were evaluated using the American Orthopaedic Foot and Ankle Society (AOFAS) scale score. We also considered the influence of scaffold type, lesion area, previous surgery, and lesion depth. the mean preoperative AOFAS scale score was 65.2 ± 13.9. The clinical results peaked at 24 months, before declining gradually to settle at a score of around 80 at the maximum follow-up of 72 months. the use of bone marrow-derived cells supported by scaffolds to repair osteochondral lesions of the talus resulted in significant clinical improvement, which was maintained over time. level IV, therapeutic case series.

  13. One-Step Preparation of Silver Hexagonal Microsheets as Electrically Conductive Adhesive Fillers for Printed Electronics.

    Science.gov (United States)

    Ren, Hu-Ming; Guo, Ying; Huang, Sheng-Yun; Zhang, Kai; Yuen, Matthew M F; Fu, Xian-Zhu; Yu, Shuhui; Sun, Rong; Wong, Ching-Ping

    2015-06-24

    A facile one-step solution-phase chemical reduction method has been developed to synthesize Ag microsheets at room temperature. The morphology of Ag sheets is a regular hexagon more than 1 μm in size and about 200 nm in thickness. The hexagonal Ag microsheets possess a smoother and straighter surface compared with that of the commercial Ag micrometer-sized flakes prepared by ball milling for electrically conductive adhesives (ECAs). The function of the reagents and the formation mechanism of Ag hexagonal microsheets are also investigated. For the polyvinylpyrrolidone (PVP) and citrate facet-selective capping, the Ag atoms freshly reduced by N2H4 would orientationally grow alone on the {111} facet of Ag seeds, with the synergistically selective etching of irregular and small Ag particles by H2O2, to form Ag hexagonal microsheets. The hexagonal Ag microsheet-filled epoxy adhesives, as electrically conductive materials, can be easily printed on various substrates such as polyethylene terephthalate (PET), epoxy, glass, and flexible papers. The hexagonal Ag microsheet filled ECAs demonstrate lower bulk resistivity (approximately 8 × 10(-5) Ω cm) than that of the traditional Ag micrometer-sized-flake-filled ECAs with the same Ag content of 80 wt % (approximately 1.2 × 10(-4) Ω cm).

  14. A Facile One Step Solution Route to Synthesize Cuprous Oxide Nanofluid

    Directory of Open Access Journals (Sweden)

    Shenoy U. Sandhya

    2013-05-01

    Full Text Available A cuprous oxide nanofluid stabilized by sodium lauryl sulfate, synthesized by using the one step method, has been reported. Nanofluids were synthesized by using a well‐ controlled surfactant‐assisted solution phase synthesis. The method involved reduction of copper acetate by glucose in a mixture of water and ethylene glycol serving as the base fluid. The synthesized fluid was characterized by X‐ray and electron diffraction techniques, in addition, transmission and field emission microscopic techniques and Fourier transform infra red spectroscopic analysis was undertaken. The rheological property, as well as the thermal conductivity of the fluid, were measured. The variation of reaction parameters considerably affected the size of the particles as well as the reaction rate. The uniform dispersion of the particles in the base fluid led to a stability period of three months under stationary state, augmenting the thermal conductivity of the nanofluid. The method is found to be simple, reliable and fast for the synthesis of Newtonian nanofluids containing cuprous oxide nanoparticles.

  15. Multiplex Reverse Transcription-Polymerase Chain Reaction for Simultaneous Screening of 29 Chromosomal Translocation in Hematologic Malignancies

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Multiplex reverse transcription-polymerase chain reaction (M-RT-PCR) has been proved to possess great clinical potential for simultaneous screening of 29 chromosomal translocations in acute leukemia. To evaluate the clinical value of M-RT-PCR in hematologic malignancies, bone marrow samples from 90 patients with various hematologic malignancies, including 25 acute myeloge nous leukemia (AML), 22 acute lymphoblastic leukemia (ALL), 27 chronic myelogenous leukemia (CML), 4 myeloproliferative diseases (MPD), 3 chronic lymphoblastic leukemia (CLL), 3 and 1 malignant histocytosis (MH) were subjected to both M-RT-PCR and chromosome karyotypic analysis. Some of cases were subjected to follow-up examination of M-RT-PCR during the period of ukemia. In our hand, 12 of 29chromosomal translocation transcripts including TEL/PDGFR, DEK/CAN, MLL/AF6, AML1/ETO,F9, BCR/ABL, MLL/MLL, PML/RARα, TLS/ERG, E2A/HLF, EVIl and HOXI1 were detected in 57 cases (63.3 %) of the 90 samples, which were in consistence with the results of karyore, M-RT-PCR had also shown good clinical relevance when used as an approach to detect minimal residual leukemia. We concluded that M-RT-PCR could be used as an effiy in the initial diagnosis of hematologic malignancies but also in subsequent monitor of minimal residual leukemia.

  16. Reverse transcription polymerase chain reaction-based method for selectively detecting vegetative cells of toxigenic Clostridium difficile.

    Science.gov (United States)

    Senoh, Mitsutoshi; Kato, Haru; Murase, Tomoko; Hagiya, Hideharu; Tagashira, Yasuaki; Fukuda, Tadashi; Iwaki, Masaaki; Yamamoto, Akihiko; Shibayama, Keigo

    2014-11-01

    The laboratory diagnostic methods for Clostridium difficile infection (CDI) include toxigenic culture, enzyme immunoassays (EIAs) to detect the toxins of C. difficile, and nucleic acid amplification tests (NAATs) to detect C. difficile toxin genes, but each of these methods has disadvantages; toxigenic cultures require a long time to produce results, EIAs have low sensitivity, and NAATs that target DNA cannot distinguish vegetative cells from spores and dead cells. Here we report a new detection method that uses reverse transcription polymerase chain reaction to target the toxin-gene transcripts. This method was able to specifically detect the vegetative cells of toxigenic C. difficile in fecal samples in spike tests, with a minimum detection limit of 5 × 10(2) colony-forming units per 100 mg of stool specimen. The performance of this method was also demonstrated in a pilot scale evaluation using clinical fecal specimens, which showed that this method may be more sensitive than EIA and requires a shorter time than toxigenic culture. This method could potentially be applied in the clinical laboratory to detect C. difficile in fecal specimens. The ability of this method to discriminate the presence of vegetative cells from spores and dead cells could help to further the understanding of CDI.

  17. Reverse transcription-polymerase chain reaction molecular testing of cytology specimens: Pre-analytic and analytic factors.

    Science.gov (United States)

    Bridge, Julia A

    2017-01-01

    The introduction of molecular testing into cytopathology laboratory practice has expanded the types of samples considered feasible for identifying genetic alterations that play an essential role in cancer diagnosis and treatment. Reverse transcription-polymerase chain reaction (RT-PCR), a sensitive and specific technical approach for amplifying a defined segment of RNA after it has been reverse-transcribed into its DNA complement, is commonly used in clinical practice for the identification of recurrent or tumor-specific fusion gene events. Real-time RT-PCR (quantitative RT-PCR), a technical variation, also permits the quantitation of products generated during each cycle of the polymerase chain reaction process. This review addresses qualitative and quantitative pre-analytic and analytic considerations of RT-PCR as they relate to various cytologic specimens. An understanding of these aspects of genetic testing is central to attaining optimal results in the face of the challenges that cytology specimens may present. Cancer Cytopathol 2017;125:11-19. © 2016 American Cancer Society.

  18. A sensitive one-step real-time PCR for detection of avian influenza viruses using a MGB probe and an internal positive control

    Directory of Open Access Journals (Sweden)

    Delogu Mauro

    2006-05-01

    Full Text Available Abstract Background Avian influenza viruses (AIVs are endemic in wild birds and their introduction and conversion to highly pathogenic avian influenza virus in domestic poultry is a cause of serious economic losses as well as a risk for potential transmission to humans. The ability to rapidly recognise AIVs in biological specimens is critical for limiting further spread of the disease in poultry. The advent of molecular methods such as real time polymerase chain reaction has allowed improvement of detection methods currently used in laboratories, although not all of these methods include an Internal Positive Control (IPC to monitor for false negative results. Therefore we developed a one-step reverse transcription real time PCR (RRT-PCR with a Minor Groove Binder (MGB probe for the detection of different subtypes of AIVs. This technique also includes an IPC. Methods RRT-PCR was developed using an improved TaqMan technology with a MGB probe to detect AI from reference viruses. Primers and probe were designed based on the matrix gene sequences from most animal and human A influenza virus subtypes. The specificity of RRT-PCR was assessed by detecting influenza A virus isolates belonging to subtypes from H1–H13 isolated in avian, human, swine and equine hosts. The analytical sensitivity of the RRT-PCR assay was determined using serial dilutions of in vitro transcribed matrix gene RNA. The use of a rodent RNA as an IPC in order not to reduce the efficiency of the assay was adopted. Results The RRT-PCR assay is capable to detect all tested influenza A viruses. The detection limit of the assay was shown to be between 5 and 50 RNA copies per reaction and the standard curve demonstrated a linear range from 5 to 5 × 108 copies as well as excellent reproducibility. The analytical sensitivity of the assay is 10–100 times higher than conventional RT-PCR. Conclusion The high sensitivity, rapidity, reproducibility and specificity of the AIV RRT-PCR with

  19. Reptin is required for the transcription of telomerase reverse transcriptase and over-expressed in gastric cancer

    Directory of Open Access Journals (Sweden)

    Liu Tiantian

    2010-05-01

    Full Text Available Abstract Background Telomerase is activated in oncogenesis, which confers an immortal phenotype to cancer cells. The AAA + ATPase Reptin is required for telomerase biogenesis by maintaining telomerase RNA (hTER stability and is aberrantly expressed in certain cancers. Given its role in chromatin remodeling and transcription regulation, we determined the effect of Reptin on the transcription of the telomerase reverse transcriptase (hTERT gene, a key component of the telomerase complex and its expression in gastric cancer. Results Knocking down Reptin or its partner Pontin using small interfering RNA in gastric and cervical cancer cells led to significant decreases in hTERT mRNA, but hTERT promoter activity was inhibited in only Reptin-depleted cells. Reptin interacted with the c-MYC oncoprotein and its stimulatory effect on the hTERTpromoter was significantly dependent on functional E-boxes in the promoter. Moreover, Reptin bound to the hTERT proximal promoter and the loss of the Reptin occupancy led to dissociation of c-MYC from the hTERT promoter in Reptin-depleted cells. Reptin inhibition dramatically impaired clonogenic potential of gastric cancer cells by inducing cell growtharrest and over-expression of Reptin was observed in primary gastric cancer specimens. Conclusions The hTERT gene is a direct target of Reptin, and hTERT transcription requires constitutive expression of Reptin and its cooperation with c-MYC. Thus, Reptin regulates telomerase at two different levels. This finding, together with the requirementof Reptin for the clonogenic potential of cancer cells and its over-expression in gastriccancer and other solid tumors, suggests that Reptin may be a putative therapeutic target.

  20. Systems analysis reveals a transcriptional reversal of the mesenchymal phenotype induced by SNAIL-inhibitor GN-25

    Science.gov (United States)

    2013-01-01

    Background HMLEs (HMLE-SNAIL and Kras-HMLE, Kras-HMLE-SNAIL pairs) serve as excellent model system to interrogate the effect of SNAIL targeted agents that reverse epithelial-to-mesenchymal transition (EMT). We had earlier developed a SNAIL-p53 interaction inhibitor (GN-25) that was shown to suppress SNAIL function. In this report, using systems biology and pathway network analysis, we show that GN-25 could cause reversal of EMT leading to mesenchymal-to-epithelial transition (MET) in a well-recognized HMLE-SNAIL and Kras-HMLE-SNAIL models. Results GN-25 induced MET was found to be consistent with growth inhibition, suppression of spheroid forming capacity and induction of apoptosis. Pathway network analysis of mRNA expression using microarrays from GN-25 treated Kras-HMLE-SNAIL cells showed an orchestrated global re-organization of EMT network genes. The expression signatures were validated at the protein level (down-regulation of mesenchymal markers such as TWIST1 and TWIST2 that was concurrent with up-regulation of epithelial marker E-Cadherin), and RNAi studies validated SNAIL dependent mechanism of action of the drug. Most importantly, GN-25 modulated many major transcription factors (TFs) such as inhibition of oncogenic TFs Myc, TBX2, NR3C1 and led to enhancement in the expression of tumor suppressor TFs such as SMAD7, DD1T3, CEBPA, HOXA5, TFEB, IRF1, IRF7 and XBP1, resulting in MET as well as cell death. Conclusions Our systems and network investigations provide convincing pre-clinical evidence in support of the clinical application of GN-25 for the reversal of EMT and thereby reducing cancer cell aggressiveness. PMID:24004452

  1. Lanthanide chelate complementation and hydrolysis enhanced luminescent chelate in real-time reverse transcription polymerase chain reaction assays for KLK3 transcripts.

    Science.gov (United States)

    Alinezhad, Saeid; Väänänen, Riina-Minna; Lehmusvuori, Ari; Karhunen, Ulla; Soukka, Tero; Kähkönen, Esa; Taimen, Pekka; Alanen, Kalle; Pettersson, Kim

    2014-01-01

    The requirement for high-performance reporter probes in real-time detection of polymerase chain reaction (PCR) has led to the use of time-resolved fluorometry of lanthanide chelates. The aim of this study was to investigate the applicability of the principle of lanthanide chelate complementation (LCC) in comparison with a method based on hydrolysis enhancement and quenching of intact probes. A real-time reverse transcription (RT) PCR assay for kallikrein-related peptidase 3 (KLK3, model analyte) was developed by using the LCC detection method. Both detection methods were tested with a standard series of purified PCR products, 20 prostatic tissues, 20 healthy and prostate cancer patient blood samples, and female blood samples spiked with LNCaP cells. The same limit of detection was obtained with both methods, and two cycles earlier detection with the LCC method was observed. KLK3 messenger RNA (mRNA) was detected in all tissue samples and in 1 of 20 blood samples identically with both methods. The background was 30 times lower, and the signal-to-background (S/B) ratio was 3 times higher, when compared with the reference method. Use of the new reporter method provided similar sensitivity and specificity as the reference method. The lower background, the improved S/B ratio, and the possibility of melting curve analysis and single nucleotide polymorphism (SNP) detection could be advantages for this new reporter probe.

  2. High rate of mismatch extension during reverse transcription in a single round of retrovirus replication.

    Science.gov (United States)

    Pulsinelli, G A; Temin, H M

    1994-09-27

    We made spleen necrosis virus-based retroviral vectors with mutations at the 3' end of the primer binding site region to observe the effects of terminal mismatches on retroviral replication. These vectors, when compared to a vector with the wild-type primer binding sequence, allowed us to assay the effects of the mutations on the viral titer during a single cycle of replication. The mutant vectors had titers that were comparable to the wild-type vector, indicating that reverse transcriptase has no trouble extending mismatches of as many as 3 bases under normal in vivo conditions. These results confirm and extend previous in vitro studies [Yu, H. & Goodman, M. (1992) J. Biol. Chem. 15, 10888-10896] that showed that such mismatch extension could occur in a cell-free system at high concentrations of incorrect nucleotides and in the absence of correct nucleotides. We now show that mismatch extension can occur during normal retroviral replication in cells and at normal physiological nucleotide concentrations.

  3. Reverse Transcription in the Saccharomyces cerevisiae Long-Terminal Repeat Retrotransposon Ty3

    Directory of Open Access Journals (Sweden)

    Jason W. Rausch

    2017-03-01

    Full Text Available Converting the single-stranded retroviral RNA into integration-competent double-stranded DNA is achieved through a multi-step process mediated by the virus-coded reverse transcriptase (RT. With the exception that it is restricted to an intracellular life cycle, replication of the Saccharomyces cerevisiae long terminal repeat (LTR-retrotransposon Ty3 genome is guided by equivalent events that, while generally similar, show many unique and subtle differences relative to the retroviral counterparts. Until only recently, our knowledge of RT structure and function was guided by a vast body of literature on the human immunodeficiency virus (HIV enzyme. Although the recently-solved structure of Ty3 RT in the presence of an RNA/DNA hybrid adds little in terms of novelty to the mechanistic basis underlying DNA polymerase and ribonuclease H activity, it highlights quite remarkable topological differences between retroviral and LTR-retrotransposon RTs. The theme of overall similarity but distinct differences extends to the priming mechanisms used by Ty3 RT to initiate (− and (+ strand DNA synthesis. The unique structural organization of the retrotransposon enzyme and interaction with its nucleic acid substrates, with emphasis on polypurine tract (PPT-primed initiation of (+ strand synthesis, is the subject of this review.

  4. Rapid detection of European orthobunyaviruses by reverse transcription loop-mediated isothermal amplification assays.

    Science.gov (United States)

    Camp, Jeremy V; Nowotny, Norbert

    2016-10-01

    The development of reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assays are described herein for the detection of two orthobunyaviruses (Bunyaviridae), which represent the two main serogroups found in mosquitoes in Central Europe. The RT-LAMP assays were optimized for the detection of Ťahyňa virus (a California encephalitis group virus found in Aedes sp or Ochlerotatus sp mosquitoes) and Batai virus (also called Čalovo virus, a Bunyamwera group virus found in Anopheles maculipennis s.l. mosquitoes) nucleic acid using endemic European virus isolates. The sensitivity of the RT-LAMP assays was determined to be comparable to that of conventional tests, with a limit of detectionisothermal conditions using very simple equipment. Furthermore, it was possible to proceed with the assays without nucleic acid extraction, albeit at a 100-fold loss of sensitivity. The RT-LAMP assays are a sensitive, cost-efficient method for both arbovirus surveillance as well as diagnostic laboratories to detect the presence of these endemic orthobunyaviruses.

  5. Development of a multiplex reverse transcription-PCR assay for simultaneous detection of garlic viruses

    Institute of Scientific and Technical Information of China (English)

    HU Xin-xi; LEI Yan; WANG Pei; TANG Lin-fei; HE Chang-zheng; SONG Yong; XIONG Xing-yao; NIE Xian-zhou

    2015-01-01

    A preliminary screening for garlic viruses in garlic plants in Hunan, China, using existing monoplex (simplex) reverse tran-scription-polymerase chain reaction (RT-PCR) procedures detected four viruses/virus groups. These viruses/virus groups were Onion yel ow dwarf virus (OYDV), Leek yel ow stripe virus (LYSV), Shal ot latent virus (SLV) and al exiviruses (e.g., garlic viruses A, B, C, D, E, X). Sequence analysis of the projected al exivirus amplicons revealed the al exivirus in the infected garlic plants was Garlic virus D (GarV-D), which shared 92–97%sequence identities with various isolates from the world. A multiplex RT-PCR (mRT-PCR) was therefore developed to simultaneously detect and differentiate the four viruses/virus groups. To achieve this, four primer pairs targeting al exiviruses, OYDV, LYSV and SLV were designed. The anticipated amplicon sizes are 183 bp (al exiviruses), 265 bp (OYDV), 404 bp (LYSV) and 592 bp (SLV), respectively. Al primer pairs produced virus-speciifc fragments in both simplex and multiplex formats, thus conifrming the efifcacy of the newly developed mRT-PCR for detection of these viruses. The mRT-PCR further was evaluated by applying it to garlic plant samples col ected in two geographic locations in Hunan. Al exiviruses, OYDV, LYSV and SLV were detected in 50.9, 40.3, 28.3 and 58.5%of leaf samples, respectively;and mixed infections with two or more viruses accounted for 54%of the garlic samples. The results obtained by mRT-PCR were conifrmed by simplex RT-PCR assays. In conclusion, this newly devel-oped mRT-PCR provides a rapid, sensitive and reliable method for the detection and identiifcation of major garlic viruses.

  6. Magnetic studies of mesoporous nanostructured iron oxide materials synthesized by one-step soft-templating.

    Science.gov (United States)

    Jin, Jing; Hines, William A; Kuo, Chung-Hao; Perry, David M; Poyraz, Altug S; Xia, Yan; Zaidi, Taha; Nieh, Mu-Ping; Suib, Steven L

    2015-07-14

    A combined magnetization and (57)Fe spin-echo nuclear magnetic resonance (NMR) study has been carried out on mesoporous nanostructured materials consisting of the magnetite (Fe3O4) and maghemite (γ-Fe2O3) phases. Two series of samples were synthesized using a recently developed one-step soft-templating approach with systematic variations in calcination temperature and reaction atmosphere. Nuclear magnetic resonance has been shown to be a valuable tool for distinguishing between the two magnetic iron oxide spinel phases, Fe3O4 and γ-Fe2O3, on the nanoscale as well as monitoring phase transformation resulting from oxidation. For the Fe3O4 and γ-Fe2O3 phases, peaks in the NMR spectra are attributed to Fe in the tetrahedral (A) sites and octahedral (B) sites. The magnetic field dependence of the peaks was observed and confirmed the site assignments. Fe3O4 on a nanoscale readily oxidizes to form γ-Fe2O3 and this was clearly evident in the NMR spectra. As evidenced by transmission electron microscope (TEM) images, the porous mesostructure for the iron oxide materials is formed by a random close-packed aggregation of nanoparticles; correspondingly, superparamagnetic behavior was observed in the magnetic measurements. Although X-ray diffraction (XRD) shows the spinel structure for the Fe3O4 and γ-Fe2O3 phases, unlike NMR, it is difficult to distinguish between the two phases with XRD. Nitrogen sorption isotherms characterize the mesoporous structures of the materials, and yield BET surface area values and limited BJH pore size distribution curves.

  7. Nanomembrane Canister Architectures for the Visualization and Filtration of Oxyanion Toxins with One-Step Processing.

    Science.gov (United States)

    Aboelmagd, Ahmed; El-Safty, Sherif A; Shenashen, Mohamed A; Elshehy, Emad A; Khairy, Mohamed; Sakaic, Masaru; Yamaguchi, Hitoshi

    2015-11-01

    Nanomembrane canister-like architectures were fabricated by using hexagonal mesocylinder-shaped aluminosilica nanotubes (MNTs)-porous anodic alumina (PAA) hybrid nanochannels. The engineering pattern of the MNTs inside a 60 μm-long membrane channel enabled the creation of unique canister-like channel necks and cavities. The open-tubular canister architecture design provides controllable, reproducible, and one-step processing patterns of visual detection and rejection/permeation of oxyanion toxins such as selenite (SeO3(2-)) in aquatic environments (i.e., in ground and river water sources) in the Ibaraki Prefecture of Japan. The decoration of organic ligand moieties such as omega chrome black blue (OCG) into inorganic Al2O3@tubular SiO2/Al2O3 canister membrane channel cavities led to the fabrication of an optical nanomembrane sensor (ONS). The OCG ligand was not leached from the canister as observed in washing, sensing, and recovery assays of selenite anions in solution, which enabled its multiple reuse. The ONS makes a variety of alternate processing analyses of selective quantification, visual detection, rejection/permeation, and recovery of toxic selenite quick and simple without using complex instrumentation. Under optimal conditions, the ONS canister exhibited a high selectivity toward selenite anions relative to other ions and a low-level detection limit of 0.0093 μM. Real analytical data showed that approximately 96% of SeO3(2-) anions can be recovered from aquatic and wastewater samples. The ONS canister holds potential for field recovery applications of toxic selenite anions from water. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. One-step electrodeposition process of CuInSe2: Deposition time effect

    Indian Academy of Sciences (India)

    O Meglali; N Attaf; A Bouraiou; M S Aida; S Lakehal

    2014-10-01

    CuInSe2 thin films were prepared by one-step electrodeposition process using a simplified twoelectrodes system. The films were deposited, during 5, 10, 15 and 20 min, from the deionized water solution consisting of CuCl2, InCl3 and SeO2 onto ITO-coated glass substrates. As-deposited films have been annealed under vacuum at 300 °C during 30 min. The structural, optical band gap and electrical resistivity of elaborated films were studied, respectively, using X-ray diffraction (XRD), Raman spectroscopy, UV spectrophotometer and four-point probe method. The micro structural parameters like lattice constants, crystallite size, dislocation density and strain have been evaluated. The XRD investigation proved that the film deposited at 20 min present CuInSe2 single phase in its chalcopyrite structure and with preferred orientation along (1 1 2) direction, whereas the films deposited at 5, 10 and 15 min show the CuInSe2 chalcopyrite structure with the In2Se3 as secondary phase. We have found that the formation mechanism of CuInSe2 depends on the In2Se3 phase. The optical band gap of the films is found to decrease from 1.17 to 1.04 eV with increase in deposition time. All films show Raman spectra with a dominant A1 mode at 174 cm-1, confirming the chalcopyrite crystalline quality of these films. The films exhibited a range of resistivity varying from 2.3 × 10-3 to 4.4 × 10-1 cm.

  9. Dramatic improvement in dissolution rate of albendazole by a simple, one-step, industrially scalable technique

    Directory of Open Access Journals (Sweden)

    Saeed Ghanbarzadeh

    2016-01-01

    Full Text Available Low solubility and dissolution rate are the primary challenges in the drug development which substantially impact the oral absorption and bioavailability of drugs. Due to the poor water solubility, Albendazole (ABZ is poorly absorbed from the gastrointestinal tract and shows low oral bioavailability (5% which is a major disadvantage for the systemic use of ABZ. To improve the solubility and dissolution rate of ABZ, different classes of hydrophilic excipients such as sugars (lactose, sucrose, and glucose, polyols (mannitol and sorbitol, ionic surfactant (sodium lauryl sulfate and non-ionic surfactant (Cremophor A25 were co-spray dried with ABZ. The crystallinity changes in the processed drug were characterized by differential scanning calorimetry and X-Ray diffraction methods were used to interpret the enhanced solubility and dissolution rate of the drug. Results showed that the solubility and dissolution rate of ABZ were increased 1.8-2.6 folds and 3-25 folds, respectively. Unexpectedly, SLS decreased the solubility index of drug powder even lower than the unprocessed drug which was attributed to drug-SLS ionic interaction as depicted from Fourier transform infrared spectroscopy. It was concluded that by applying the facile, one-step, industrially scalable technique and the use of small amounts of excipient (only 4% of the formulation, a great improvement (21 folds in dissolution rate of ABZ was achieved. This finding may be used in the pharmaceutical industries for the formulation of therapeutically efficient dosage forms of class II and IV drugs classified in biopharmaceutical classification system.

  10. Development of a One-Step Multiplex PCR Assay for Differential Detection of Major Mycobacterium Species.

    Science.gov (United States)

    Chae, Hansong; Han, Seung Jung; Kim, Su-Young; Ki, Chang-Seok; Huh, Hee Jae; Yong, Dongeun; Koh, Won-Jung; Shin, Sung Jae

    2017-09-01

    The prevalence of tuberculosis continues to be high, and nontuberculous mycobacterial (NTM) infection has also emerged worldwide. Moreover, differential and accurate identification of mycobacteria to the species or subspecies level is an unmet clinical need. Here, we developed a one-step multiplex PCR assay using whole-genome analysis and bioinformatics to identify novel molecular targets. The aims of this assay were to (i) discriminate between the Mycobacterium tuberculosis complex (MTBC) and NTM using rv0577 or RD750, (ii) differentiate M. tuberculosis (M. tuberculosis) from MTBC using RD9, (iii) selectively identify the widespread M. tuberculosis Beijing genotype by targeting mtbk_20680, and (iv) simultaneously detect five clinically important NTM (M. avium, M. intracellulare, M. abscessus, M. massiliense, and M. kansasii) by targeting IS1311, DT1, mass_3210, and mkan_rs12360 An initial evaluation of the multiplex PCR assay using reference strains demonstrated 100% specificity for the targeted Mycobacterium species. Analytical sensitivity ranged from 1 to 10 pg for extracted DNA and was 10(3) and 10(4) CFU for pure cultures and nonhomogenized artificial sputum cultures, respectively, of the targeted species. The accuracy of the multiplex PCR assay was further evaluated using 55 reference strains and 94 mycobacterial clinical isolates. Spoligotyping, multilocus sequence analysis, and a commercial real-time PCR assay were employed as standard assays to evaluate the multiplex PCR assay with clinical M. tuberculosis and NTM isolates. The PCR assay displayed 100% identification agreement with the standard assays. Our multiplex PCR assay is a simple, convenient, and reliable technique for differential identification of MTBC, M. tuberculosis, M. tuberculosis Beijing genotype, and major NTM species. Copyright © 2017 American Society for Microbiology.

  11. One step synthesis of polyacrylamide functionalized graphene and its application in Pb(II) removal

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Zhiwei; Zhang, Yaoyao; Qian, Xiaoming; Shi, Jie; Chen, Lei; Li, Baodong; Niu, Jiarong; Liu, Liangsen, E-mail: 83019163@163.com

    2014-10-15

    Highlights: • PAM-g-graphene is synthesized by the co-irradiation between GO and AM monomers. • PAM graft on GO has led to the exfoliation of GO into individual sheets. • The γ-ray induced reduction of GO. • PAM-g-graphene exhibits high adsorption capacities toward Pb(II) ions. • PAM-g-graphene provides a new idea for heavy metal pollutants’ removal in water. - Abstract: Polyacrylamide grafted graphene (PAM-g-graphene) from graphite oxide (GO) was successfully prepared by γ-ray irradiation with acrylamide monomers in aqueous at room temperature in this paper. Our strategy involves the PAM chains graft on the surface and between the layers of GO by in situ radical polymerization which led to the exfoliation of GO into individual sheets. Results show that the degree of grafting of PAM-g-graphene samples is 24.2%, and the thickness is measured to be 2.59 nm. Moreover, the as-prepared PAM-g-graphene with some amino from PAM and little oxygen functional groups exhibit superior adsorption of Pb(II) ions. The adsorption processes reach equilibrium in just 30 min and the adsorption isotherms are described well by Langmuir and Freundlich classical isotherms models. The determined adsorption capacity of PAM-g-graphene is 819.67 mg g{sup −1} (pH 6) for Pb(II), which is 20 times and 8 times capacities of that for graphene nanosheets and carbon nanotubes according to reports, respectively. This chemically modified graphene synthesized by this fast one-step approach, featuring a good versatility and adaptability, excellent adsorption capacity and rapid extraction, may provide a new idea for the global problem of heavy metal pollutants’ removal in water.

  12. Synthesis of carbon nanoparticles using one step green approach and their application as mercuric ion sensor

    Energy Technology Data Exchange (ETDEWEB)

    Roshni, V.; Ottoor, Divya, E-mail: divya@chem.unipune.ac.in

    2015-05-15

    Carbon nanoparticles (CNPs) have been evolved as a promising candidate for the metal sensing applications due to their synthesis from naturally occurring and easily available non-toxic molecular precursors by green chemistry. A simple and one step procedure is reported here for the synthesis of CNPs from coconut milk by thermal pyrolysis at a temperature of 120–150 °C for 2–5 min without using any carbonizing or passivating agent. On pyrolysis the coconut oil is separated from the carbon rich residue and the residue when dissolved in water showed blue fluorescence under UV light. The CNPs produced are found to show an emission maximum at 440 nm when excited at 360 nm. Synthesis by green approach makes CNPs a promising substitute for the metal sensing applications. Series of metal ions which have a hazardous impact on the ecological system have been taken for the analysis and it is observed that the fluorescence of CNPs gets remarkably quenched by mercuric ions. Fluorescence quenching was studied using standard Stern–Volmer quenching model. Limit of detection was found to be 16.5 nM Hg{sup 2+} concentration. - Highlights: • Green and economical synthesis of carbon nanoparticles (CNPs) from naturally abundant material. • Coconut milk is used as molecular precursor, which on thermal pyrolysis at 120 °C yielded CNPs. • Highly fluorescent CNPs show an emission maxima of 440 nm when excited at 360 nm. • Application of CNPs for metal ion sensing using fluorescence quenching phenomena. • Hg{sup 2+} is most effectively sensed with a detection limit of 16.5 nM.

  13. One-step production of immobilized alpha-amylase in recombinant Escherichia coli.

    Science.gov (United States)

    Rasiah, Indira A; Rehm, Bernd H A

    2009-04-01

    Industrial enzymes are often immobilized via chemical cross-linking onto solid supports to enhance stability and facilitate repeated use in bioreactors. For starch-degrading enzymes, immobilization usually places constraints on enzymatic conversion due to the limited diffusion of the macromolecular substrate through available supports. This study describes the one-step immobilization of a highly thermostable alpha-amylase (BLA) from Bacillus licheniformis and its functional display on the surface of polyester beads inside engineered Escherichia coli. An optimized BLA variant (Termamyl) was N-terminally fused to the polyester granule-forming enzyme PhaC of Cupriavidus necator. The fusion protein lacking the signal sequence mediated formation of stable polyester beads exhibiting alpha-amylase activity. The alpha-amylase beads were assessed with respect to alpha-amylase activity, which was demonstrated qualitatively and quantitatively. The immobilized alpha-amylase showed Michaelis-Menten enzyme kinetics exerting a V(max) of about 506 mU/mg of bead protein with a K(m) of about 5 microM, consistent with that of free alpha-amylase. The stability of the enzyme at 85 degrees C and the capacity for repeated usage in a starch liquefaction process were also demonstrated. In addition, structural integrity and functionality of the beads at extremes of pH and temperature, demonstrating their suitability for industrial use, were confirmed by electron microscopy and protein/enzyme analysis. This study proposes a novel, cost-effective method for the production of immobilized alpha-amylase in a single step by using the polyester granules forming protein PhaC as a fusion partner in engineered E. coli.

  14. One-step extraction of polar drugs from plasma by parallel artificial liquid membrane extraction.

    Science.gov (United States)

    Pilařová, Veronika; Sultani, Mumtaz; Ask, Kristine Skoglund; Nováková, Lucie; Pedersen-Bjergaard, Stig; Gjelstad, Astrid

    2017-02-01

    The new microextraction technique named parallel artificial liquid membrane extraction (PALME) was introduced as an alternative approach to liquid-liquid extraction of charged analytes from aqueous samples. The concept is based on extraction of analytes across a supported liquid membrane sustained in the pores of a thin polymeric membrane, a well-known extraction principle also used in hollow fiber liquid-phase microextraction (HF-LPME). However, the new PALME technique offers a more user-friendly setup in which the supported liquid membrane is incorporated in a 96 well plate system. Thus, high-throughput is achievable, in addition to the green chemistry offered by using PALME. The consumption of organic solvent is minimized to 3-5μL per sample. With a sample volume of 250μL and acceptor solution volume of 50μL, a maximal enrichment factor of five is achievable. Based on these parameters, a new method for extraction of polar basic drugs was developed in the present work. The basic drugs hydralazine, ephedrine, metaraminol, salbutamol, and cimetidine were used as model analytes, and were extracted from alkalized human plasma into an aqueous solution via the supported liquid membrane. The extraction was promoted by a carrier dissolved in the membrane, creating a temporary ion-pair complex between the hydrophilic drug and the carrier. As the model analytes were extracted directly into an aqueous solution, there was no need for evaporation of the extract before injection into LC-MS. Hence, the sample preparation is performed in one step. With optimized conditions, the extraction recoveries were in the range 50-89% from human plasma after 45min extraction. The data from the method evaluation were satisfactory and in line with current guidelines, and revealed an extraction method with substantial potential for high throughput bioanalysis of polar basic drugs.

  15. One-step high-efficiency CRISPR/Cas9-mediated genome editing in Streptomyces.

    Science.gov (United States)

    Huang, He; Zheng, Guosong; Jiang, Weihong; Hu, Haifeng; Lu, Yinhua

    2015-04-01

    The RNA-guided DNA editing technology CRISPRs (clustered regularly interspaced short palindromic repeats)/Cas9 had been used to introduce double-stranded breaks into genomes and to direct subsequent site-specific insertions/deletions or the replacement of genetic material in bacteria, such as Escherichia coli, Streptococcus pneumonia, and Lactobacillus reuteri. In this study, we established a high-efficiency CRISPR/Cas9 genome editing plasmid pKCcas9dO for use in Streptomyces genetic manipulation, which comprises a target-specific guide RNA, a codon-optimized cas9, and two homology-directed repair templates. By delivering pKCcas9dO series editing plasmids into the model strain Streptomyces coelicolor M145, through one-step intergeneric transfer, we achieved the genome editing at different levels with high efficiencies of 60%-100%, including single gene deletion, such as actII-orf4, redD, and glnR, and single large-size gene cluster deletion, such as the antibiotic biosynthetic clusters of actinorhodin (ACT) (21.3 kb), undecylprodigiosin (RED) (31.6 kb), and Ca(2+)-dependent antibiotic (82.8 kb). Furthermore, we also realized simultaneous deletions of actII-orf4 and redD, and of the ACT and RED biosynthetic gene clusters with high efficiencies of 54% and 45%, respectively. Finally, we applied this system to introduce nucleotide point mutations into the rpsL gene, which conferred the mutants with resistance to streptomycin. Notably, using this system, the time required for one round of genome modification is reduced by one-third or one-half of those for conventional methods. These results clearly indicate that the established CRISPR/Cas9 genome editing system substantially improves the genome editing efficiency compared with the currently existing methods in Streptomyces, and it has promise for application to genome modification in other Actinomyces species.

  16. One-Step Synthesis of Zeolite Membranes Containing Catalytic Metal Nanoclusters.

    Science.gov (United States)

    Kim, Seok-Jhin; Tan, Shuai; Taborga Claure, Micaela; Briones Gil, Laura; More, Karren L; Liu, Yujun; Moore, Jason S; Dixit, Ravindra S; Pendergast, John G; Sholl, David S; Jones, Christopher W; Nair, Sankar

    2016-09-21

    Metal-loaded zeolitic membranes are promising candidates as catalytic membrane reactors. We report a one-step synthesis method to synthesize zeolite membranes containing metal nanoclusters, that has advantages in comparison to multistep methods such as impregnation and ion exchange. Pure-silica MFI zeolite-Pt hybrid membranes were prepared by hydrothermal synthesis with addition of 3-mercaptopropyl-trimethoxysilane (MPS) and a platinum precursor. Composition analysis and mapping by energy-dispersive X-ray spectroscopy (EDX) reveal that Pt ions/clusters are uniformly distributed along the membrane cross-section. High-magnification scanning transmission electron microscopy (STEM) analysis shows that Pt metal clusters in the hybrid zeolite membrane have a diameter distribution in the range of 0.5-2.0 nm. In contrast, a pure-silica MFI membrane synthesized from an MPS-free solution shows negligible incorporation of Pt metal clusters. To characterize the properties of the hybrid (zeolite/metal) membrane, it was used as a catalytic membrane reactor (CMR) for high-temperature propane dehydrogenation (PDH) at 600 °C and 1 atm. The results indicate that Pt metal clusters formed within the MFI zeolite membrane can serve as effective catalysts for high-temperature PDH reaction along with H2 removal via membrane permeation, thereby increasing both conversion and selectivity in relation to a conventional membrane reactor containing an equivalent amount of packed Pt catalyst in contact with an MFI membrane. The hybrid zeolite-Pt CMR also showed stable conversion and selectivity upon extended high-temperature operation (12 h), indicating that encapsulation in the zeolite allowed thermal stabilization of the Pt nanoclusters and reduced catalyst deactivation.

  17. Vygotsky's Principle "One Step in Learning - One Hundred Steps In Development": From Idea To Practice

    Directory of Open Access Journals (Sweden)

    Zaretsky V.K.,

    2016-12-01

    Full Text Available The article reviews Lev Vygotsky’s published works to trace the evolution of his understanding of child development. The authors believe that his assumption that one step in learning may mean one hundred steps in development, is as important as the two other key postulates of the cultural-historical theory: the principle that learning precedes development and the concept of zone of proximal development. The authors provide a rationale for utilization of these assumptions in the practice of development-facilitating psychological and educational assistance. A mechanism of this learning-development relationship is hypothesized. The article outlines a multidimensional model of the zone of proximal development illustrating the above mechanism. This model is one of the conceptual tools of the Reflection and Activity Approach helping children overcome learning difficulties and promoting their development. Having given the account of how they proceeded “from the idea to the problem” and “from the idea to the mechanism”, the authors provide case studies showing how this mechanism allows working with learning difficulties to trigger simultaneous improvement in multiple developmental dimensions. The article reports on the experience of running special Summer Schools for children with learning difficulties, implementing the “Chess for General Development” Project, and assisting orphaned children with severe somatic conditions. A case study of a female college student displaying signs of the learned helplessness syndrome is presented. The authors infer that Vygotsky’s idea of a specific relationship between learning and development may be of fundamental theoretical and practical value, especially for working with children with special needs.

  18. Development of a one-step duplex RT-qPCR for the quantification of phocine distemper virus.

    Science.gov (United States)

    Bogomolni, Andrea L; Frasca, Salvatore; Matassa, Keith A; Nielsen, Ole; Rogers, Kara; De Guise, Sylvain

    2015-04-01

    Worldwide, stranded marine mammals and the network personnel who respond to marine mammal mortality have provided much of the information regarding marine morbillivirus infections. An assay to determine the amount of virus present in tissue samples would be useful to assist in routine surveying of animal health and for monitoring large-scale die-off events. False negatives from poor-quality samples prevent determination of the true extent of infection, while only small amounts of tissue samples or archived RNA may be available at the time of collection for future retrospective analysis. We developed a one-step duplex real-time reverse transcriptase-quantitative-PCR assay (RT-qPCR) based on Taqman probe technology to quantify phocine distemper virus (PDV) isolated from an outbreak in harbor (Phoca vitulina concolor) and gray seals (Halichoerus grypus) along the northeast US coast in 2006. The glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) gene was selected to assess RNA quality. This duplex assay is specific for PDV and sensitive through a range of 10(0) to 10(9) copies ds-plasmid DNA. For the GAPDH target, the reaction in duplex amplified 10(0) to 10(9) copies of ds-plasmid DNA and was detectable in multiple seal species. This assay reduced the likelihood of false negative results due to degradation of tissues and well-to-well variability while providing sensitive and specific detection of PDV, which would be applicable in molecular epidemiologic studies and pathogen detection in field and laboratory investigations involving a variety of seal species.

  19. Rapid detection of tdh and trh mRNAs of Vibrio parahaemolyticus by the transcription-reverse transcription concerted (TRC) method.

    Science.gov (United States)

    Masuda, Noriyoshi; Yasukawa, Kiyoshi; Isawa, Yuichi; Horie, Ryuichi; Saitoh, Juichi; Ishiguro, Takahiko; Nakaguchi, Yoshitsugu; Nishibuchi, Mitsuaki; Hayashi, Toshinori

    2004-01-01

    We developed a novel method named the transcription-reverse transcription concerted (TRC) method and an instrument that allowed rapid and completely homogeneous real-time monitoring of RNA isothermal sequence amplification without any post-amplification analysis in our previous study [Ishiguro et al., Anal. Biochem., 314, 77-86 (2003)]. In this study, we newly established rapid and sensitive TRC systems for the detection of the mRNAs transcribed from two major virulence genes of Vibrio parahaemolyticus: the tdh gene encoding the thermostable direct hemolysin (tdh) and the trh gene encoding the thermostable direct hemolysin-related hemolysin. Examination of the standard RNAs prepared in vitro showed that a positive result, increase in the fluorescence intensity to the cut-off value within 25 min, was obtained for as few as 100 copies of RNA. The TRC method specific to the trh mRNA detected the mRNAs transcribed from the trh1 and trh2 genes, two representative trh variants sharing 84% sequence identity. The detection time gave a linear relationship to the logarithm of starting RNA copies ranging from 10(3) to 10(7) copies, showing that quantitative analysis is possible. The detection time for 10(3) copies of the standard RNAs ranged from 11 to 15 min. Examination of the total RNAs extracted from the standard strains of V. parahaemolyticus demonstrated that the new TRC systems are sufficiently sensitive to detect as few as 100 CFUs of the strains carrying the target genes. Total RNA preparations extracted from 24 strains of V. parahaemolyticus, 52 strains belonging to 31 other species of the genus Vibrio and 11 strains belonging to 8 species of non-Vibrio genera were examined. The results of the detection of tdh- and trh-specific mRNAs by the two TRC systems and those of the respective genes by the DNA colony hybridization method agreed. We conclude that the new TRC systems are rapid, highly sensitive, easy to manipulate, and are suitable for routine examination of

  20. Capillary-driven multiparametric microfluidic chips for one-step immunoassays.

    Science.gov (United States)

    Gervais, Luc; Hitzbleck, Martina; Delamarche, Emmanuel

    2011-09-15

    Here we present a capillary-driven microfluidic chip for "one-step" immunoassays. The chip allows for easy modification of several assay parameters such as the flow rates of sample, the volumes of samples for tests, and the type of reagents and receptors for detecting analytes. We therefore term such a chip a multiparametric chip and illustrate this concept with the integration and release of anti-C-reactive protein (CRP) detection antibodies (dAbs) together with splitting flow of samples containing CRP across lines of anti-CRP capture antibodies (cAbs). The microfluidic chip is fabricated in Si and is sealed with polydimethylsiloxane (PDMS) patterned with cAbs. The microfluidic chip is ∼1.7×3.4 cm(2) and is capable of analyzing 20 μL of human serum in 6 parallel flow paths with a range of flow rates from 3.3 nL s(-1) to 0.46 nL s(-1). An inkjet spotter was used to deposit 10.6 nL of dAb solution in a structure vicinal to the main flow path of the chip. The consequent asymmetric release of dAbs in a stream of human serum is compensated by a Dean flow mixer having 9 mixing loops and a footprint of 2.8 mm × 0.78 mm. The quantity of dAb present in the half of the flow path close to the spotting region decreases from 83% at the entrance of the mixer to 52% in the region after the mixer. The sample is then equally split into 6 reaction chambers and proceeds via connecting channels to 2 μL capillary pumps. The hydraulic resistance of the connecting channels is designed to vary flow rates, and therefore the kinetics of capture of CRP-dAb complexes, from 10 min to 72 min. The increased incubation time leads to a fourfold increase in detection signal in the reaction chamber with the longer incubation time. The concept presented here is flexible and suited for implementing various surface fluorescence immunoassays on a capillary-driven microfluidic chip.

  1. A new deterministic Ensemble Kalman Filter with one-step-ahead smoothing for storm surge forecasting

    KAUST Repository

    Raboudi, Naila

    2016-11-01

    The Ensemble Kalman Filter (EnKF) is a popular data assimilation method for state-parameter estimation. Following a sequential assimilation strategy, it breaks the problem into alternating cycles of forecast and analysis steps. In the forecast step, the dynamical model is used to integrate a stochastic sample approximating the state analysis distribution (called analysis ensemble) to obtain a forecast ensemble. In the analysis step, the forecast ensemble is updated with the incoming observation using a Kalman-like correction, which is then used for the next forecast step. In realistic large-scale applications, EnKFs are implemented with limited ensembles, and often poorly known model errors statistics, leading to a crude approximation of the forecast covariance. This strongly limits the filter performance. Recently, a new EnKF was proposed in [1] following a one-step-ahead smoothing strategy (EnKF-OSA), which involves an OSA smoothing of the state between two successive analysis. At each time step, EnKF-OSA exploits the observation twice. The incoming observation is first used to smooth the ensemble at the previous time step. The resulting smoothed ensemble is then integrated forward to compute a "pseudo forecast" ensemble, which is again updated with the same observation. The idea of constraining the state with future observations is to add more information in the estimation process in order to mitigate for the sub-optimal character of EnKF-like methods. The second EnKF-OSA "forecast" is computed from the smoothed ensemble and should therefore provide an improved background. In this work, we propose a deterministic variant of the EnKF-OSA, based on the Singular Evolutive Interpolated Ensemble Kalman (SEIK) filter. The motivation behind this is to avoid the observations perturbations of the EnKF in order to improve the scheme\\'s behavior when assimilating big data sets with small ensembles. The new SEIK-OSA scheme is implemented and its efficiency is demonstrated

  2. Patient satisfaction with a novel one-step hydrogen peroxide solution

    Directory of Open Access Journals (Sweden)

    Schafer J

    2014-10-01

    Full Text Available Jeffery Schafer, Robert Steffen, Marjorie J Rah Bausch & Lomb Incorporated, Rochester, NY, USA Purpose: We aimed to evaluate the product performance of a novel one-step hydrogen ­peroxide cleaning and disinfecting solution, PeroxiClear (“Test” solution, when used by habitual Clear Care users to bilaterally clean and disinfect their soft contact lenses, for approximately 2 weeks.Methods: This was a 2-week, open-label, bilateral eye study designed to include subjects ­ranging in age from 18 to 55 years, inclusive. All subjects were habitual users of the Clear Care peroxide regimen for cleaning, disinfecting, and storage of their soft contact lenses, for at least 6 months prior to enrolling in the study. Subjects were examined at two study visits: a screening/­dispensing visit and a 2-week follow-up visit. The primary end point, patient preference for the Test solution, was evaluated with an online survey administered after 7 days of using the Test cleaning and disinfecting solution. Respondents could answer questions with neutral or nonneutral responses (better or worse. Statistical analyses were conducted to compare differences for nonneutral responses.Results: Of the 299 eligible subjects enrolled, 297 completed the study, conducted at 21 sites by 21 investigators in the United States. A significantly higher proportion of nonneutral respondents reported the Test solution was better overall (85.9% than their habitual contact lens solution (14.1% (P<0.001. The proportion of subjects who preferred the Test solution over their habitual solution was significantly higher for each of the preference questions regarding comfort (85.4% vs 14.6%, moistness (90.0% vs 10.0%, cleanness (91.6% vs 8.4%, and clarity of vision (85.8% vs 14.2%.Conclusion: After 7 days of using the Test cleaning and disinfecting solution, survey results indicated high levels of patient satisfaction and preference over the habitual solution, particularly in the areas of

  3. Design and Assessment of a Real Time Reverse Transcription-PCR Method to Genotype Single-Stranded RNA Male-Specific Coliphages (Family Leviviridae).

    Science.gov (United States)

    A real-time, reverse transcription-PCR (RT-qPCR) assay was developed to differentiate the four genogroups of male-specific ssRNA coliphages (FRNA) (family Leviviridae). As FRNA display a trend of source-specificity (human sewage or animal waste) at the genogroup level, this assa...

  4. A bacterial community analysis using reverse transcription (RT) PCR which detects the bacteria with high activity in a wastewater treatment reactor

    Science.gov (United States)

    This research used reverse transcription polymerase chain reaction (RT-PCR) method to help detect active bacteria in a single-tank deammonification reactor combining partial nitritation and anammox. The single-tank aerobic deammonification reactor effectively removed the ammonia in anaerobically di...

  5. Detection of dengue virus serotype 3 by reverse transcription-polymerase chain reaction in Aedes aegypti (Diptera, Culicidae captured in Manaus, Amazonas

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    Valéria CS Pinheiro

    2005-12-01

    Full Text Available The detection of dengue virus serotypes from Aedes aegypti in Manaus, state of Amazonas was carried out using the reverse transcription-polymerase chain reaction technique. Fourteen pools out 82 (17.1% were positive for DENV3, providing a minimal infection rate of 2.1% of all analyzed infected female specimens of three different areas of the city.

  6. Development of reverse transcription loop-mediated isothermal amplification assay for rapid detection of an emerging potyvirus: tomato necrotic stunt virus

    Science.gov (United States)

    Tomato necrotic stunt virus (ToNStV) is an emerging potyvirus that causes severe stunting to the infected tomato plants. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for a sensitive detection of ToNStV. The sensitivity of RT-LAMP was comparable to th...

  7. Development of a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of Sugarcane mosaic virus and Sorghum mosaic virus in sugarcane

    Science.gov (United States)

    A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detecting Sugarcane mosaic virus (SCMV) and Sorghum mosaic virus (SrMV) in sugarcane. Six sets of four primers corresponding to the conserved coat protein gene were designed for each virus and their succ...

  8. Development of a rapid diagnostic assay for the detection of tomato chlorotic dwarf viroid based on isothermal reverse-transcription-recombinase polymerase amplification

    Science.gov (United States)

    A molecular diagnostic assay utilizing reverse transcription-recombinase polymerase amplification (RT-RPA) at an isothermal constant temperature of 39 °C and target-specific primers and probe were developed for the rapid, sensitive, and specific detection of tomato chlorotic dwarf viroid (TCDVd) in ...

  9. Multiplex Reverse Transcription-Polymerase Chain Reaction untuk Deteksi Cepat Virus Flu Burung H5N1 (MULTIPLEX REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION FOR RAPID DETECTION OF H5N1 AVIAN INFLUENZA VIRUS

    Directory of Open Access Journals (Sweden)

    Raden Wasito

    2015-05-01

    Full Text Available Avian influenza virus subtype H5N1 (AIV H5N1 is highly pathogenic and fatal in poultry. The virusis still endemic with low virulence rate, although it may play a critical role in causing high morbidity andmortality rates in poultry in Indonesia. In general, diagnostic approach for AIV H5N1 is based onconventional serological and viral isolation methods that have the potential to produce consumings oftime and relatively expensive cost within the laboratory without compromising test utility. Thus, amolecular approach of multiplex reverse transcription-polymerase chain reaction (mRT-PCR was developedand applied for the detection of matrix gene type A influenza viruses, AIV subtype subtype H5hemagglutinin gene with simultaneous detection of N1 nucleoprotein gene. Thirty sera specimens fromthe diseased commercial chickens that were specifically amplified positive-RT-PCR for AIV H5N1 wereselected for mRT-PCR. The mRT-PCR products were visualized by agarose gel electrophoresis and consistedof DNA fragments of AIV of 245 bp, 545 bp and 343 bp for M, H5 and N1 genes, respectively. Thus, themRT-PCR that can rapidly differentiate simultaneously between these genes is very important for thecontrol and even eradication of AIV transmission in poultry in Indonesia.

  10. Optimization of the elution buffer and concentration method for detecting hepatitis E virus in swine liver using a nested reverse transcription-polymerase chain reaction and real-time reverse transcription-polymerase chain reaction.

    Science.gov (United States)

    Son, Na Ry; Seo, Dong Joo; Lee, Min Hwa; Seo, Sheungwoo; Wang, Xiaoyu; Lee, Bog-Hieu; Lee, Jeong-Su; Joo, In-Sun; Hwang, In-Gyun; Choi, Changsun

    2014-09-01

    The aim of this study was to develop an optimal technique for detecting hepatitis E virus (HEV) in swine livers. Here, three elution buffers and two concentration methods were compared with respect to enhancing recovery of HEV from swine liver samples. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and nested RT-PCR were performed to detect HEV RNA. When phosphate-buffered saline (PBS, pH 7.4) was used to concentrate HEV in swine liver samples using ultrafiltration, real-time RT-PCR detected HEV in 6 of the 26 samples. When threonine buffer was used to concentrate HEV using polyethylene glycol (PEG) precipitation and ultrafiltration, real-time RT-PCR detected HEV in 1 and 3 of the 26 samples, respectively. When glycine buffer was used to concentrate HEV using ultrafiltration and PEG precipitation, real-time RT-PCR detected HEV in 1 and 3 samples of the 26 samples, respectively. When nested RT-PCR was used to detect HEV, all samples tested negative regardless of the type of elution buffer or concentration method used. Therefore, the combination of real-time RT-PCR and ultrafiltration with PBS buffer was the most sensitive and reliable method for detecting HEV in swine livers. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Rapid and Sensitive Salmonella Typhi Detection in Blood and Fecal Samples Using Reverse Transcription Loop-Mediated Isothermal Amplification.

    Science.gov (United States)

    Fan, Fenxia; Yan, Meiying; Du, Pengcheng; Chen, Chen; Kan, Biao

    2015-09-01

    Typhoid fever caused by Salmonella enterica serovar Typhi remains a significant public health problem in developing countries. Although the main method for diagnosing typhoid fever is blood culture, the test is time consuming and not always able to detect infections. Thus, it is very difficult to distinguish typhoid from other infections in patients with nonspecific symptoms. A simple and sensitive laboratory detection method remains necessary. The purpose of this study is to establish and evaluate a rapid and sensitive reverse transcription-based loop-mediated isothermal amplification (RT-LAMP) method to detect Salmonella Typhi infection. In this study, a new specific gene marker, STY1607, was selected to develop a STY1607-RT-LAMP assay; this is the first report of specific RT-LAMP detection assay for typhoid. Human-simulated and clinical blood/stool samples were used to evaluate the performance of STY1607-RT-LAMP for RNA detection; this method was compared with STY1607-LAMP, reverse transcription real-time polymerase chain reaction (rRT-PCR), and bacterial culture methods for Salmonella Typhi detection. Using mRNA as the template, STY1607-RT-LAMP exhibited 50-fold greater sensitivity than STY1607-LAMP for DNA detection. The STY1607-RT-LAMP detection limit is 3 colony-forming units (CFU)/mL for both the pure Salmonella Typhi samples and Salmonella Typhi-simulated blood samples and was 30 CFU/g for the simulated stool samples, all of which were 10-fold more sensitive than the rRT-PCR method. RT-LAMP exhibited improved Salmonella Typhi detection sensitivity compared to culture methods and to rRT-PCR of clinical blood and stool specimens from suspected typhoid fever patients. Because it can be performed without sophisticated equipment or skilled personnel, RT-LAMP is a valuable tool for clinical laboratories in developing countries. This method can be applied in the clinical diagnosis and care of typhoid fever patients as well as for a quick public health response.

  12. Identification of valid reference genes for gene expression studies of human stomach cancer by reverse transcription-qPCR

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    Lee Yeon-Su

    2010-05-01

    Full Text Available Abstract Background Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR is a powerful method for the analysis of gene expression. Target gene expression levels are usually normalized to a consistently expressed reference gene also known as internal standard, in the same sample. However, much effort has not been expended thus far in the search for reference genes suitable for the study of stomach cancer using RT-qPCR, although selection of optimal reference genes is critical for interpretation of results. Methods We assessed the suitability of six possible reference genes, beta-actin (ACTB, glyceraldehydes-3-phosphate dehydrogenase (GAPDH, hypoxanthine phosphoribosyl transferase 1 (HPRT1, beta-2-microglobulin (B2M, ribosomal subunit L29 (RPL29 and 18S ribosomal RNA (18S rRNA in 20 normal and tumor stomach tissue pairs of stomach cancer patients and 6 stomach cancer cell lines, by RT-qPCR. Employing expression stability analyses using NormFinder and geNorm algorithms we determined the order of performance of these reference genes and their variation values. Results This RT-qPCR study showed that there are statistically significant (p Conclusion This study validated RPL29 and RPL29-B2M as the best single reference genes and combination, for RT-qPCR analysis of 'all stomach tissues', and B2M and B2M-GAPDH as the best single reference gene and combination, for 'stomach cancer cell lines'. Use of these validated reference genes should provide more exact interpretation of differential gene expressions at transcription level in stomach cancer.

  13. [Colorimetric detection of human influenza A H1N1 virus by reverse transcription loop mediated isothermal amplification].

    Science.gov (United States)

    Nie, Kai; Wang, Da-Yan; Qin, Meng; Gao, Rong-Bao; Wang, Miao; Zou, Shu-Mei; Han, Feng; Zhao, Xiang; Li, Xi-Yan; Shu, Yue-Long; Ma, Xue-Jun

    2010-03-01

    A simple, rapid and sensitive colorimetric Reverse Transcription Loop Mediated Isothermal Amplification (RT-LAMP) method was established to detect human influenza A H1N1 virus. The method employed a set of six specially designed primers that recognized eight distinct sequences of the HA gene for amplification of nucleic acid under isothermal conditions at 65 degrees C for one and half hour. The amplification process of RT-LAMP was monitored by the addition of HNB (Hydroxy naphthol blue) dye prior to amplification. A positive reaction was indicated by a color change from violet to sky blue and confirmed by agarose electrophoresis. The specificity of the RT-LAMP assay was validated by cross-reaction with different swine and human influenza virus including human seasonal influenza A /H1N1 A /H3N2, influenza B and swine A /H1N1. The sensitivity of this assay was evaluated by serial dilutions of RNA molecules from in vitro transcription of human influenza A H1N1 HA gene. The assay was further evaluated with 30 clinical specimens with suspected pandemic influenza A H1N1 virus infection in parallel with RT-PCR detection and 26 clinical specimens with seasonal influenza virus infection. Our results showed that the RT-LAMP was able to achieve a sensitivity of 60 RNA copies with high specificity, and detection rate was comparable to that of the RT-PCR with the clinical samples of pandemic influenza A H1N1 infection. The RT-LAMP reaction with HNB could also be measured at 650nm in a microplate reader for quantitative analysis. Thus, we concluded that this colorimetric RT-LAMP assay had potential for the rapid screening of the human influenza A H1N1 virus infection in National influenza monitoring network laboratories and sentinel hospitals of provincial and municipal region in China.

  14. Lab-on-a-chip mRNA purification and reverse transcription via a solid-phase gene extraction technique.

    Science.gov (United States)

    Nestorova, Gergana G; Hasenstein, Karl; Nguyen, Nam; DeCoster, Mark A; Crews, Niel D

    2017-03-14

    Extraction and purification of high quality RNA is a crucial initial step required for a variety of genomic assays. We report a solid phase gene extraction (SPGE) method for automated extraction, purification and reverse transcription of mRNA in a microfluidic device. This is performed using a 130 μm diameter stainless steel needle that is amino-linked to dT(15) oligonucleotides for selective hybridization of mRNA. By inserting this probe into the biological sample for only 30 seconds, mRNA is captured with high selectivity and a yield greater than 10 pg per mm of probe length. The probe is then inserted into a lab-on-a-chip device, where the bound poly-adenylated RNA is thermally released and immediately reverse transcribed for subsequent PCR amplification. The insertion of the probe into the microfluidic device is straightforward: the microchannel is formed with an elastomer (PDMS) that, when punctured, will seal around the probe. The specificity and RNA loading capacity of the probes were evaluated using conventional qPCR. This procedure was successfully used to extract, purify, and transcribe mRNA from rat glioblastoma cell spheroids in less than seven minutes. Analysis of the product confirmed that the SPGE technique selectively captures and inherently purifies high-quality mRNA directly from biological material with no need for additional pre-processing steps. Integrating this elegant sample preparation method into a complete lab-on-a-chip system will substantially enhance the speed and automation of mRNA assays for research and clinical diagnostics.

  15. Rapid screening of innate immune gene expression in zebrafish using reverse transcription - multiplex ligation-dependent probe amplification

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    Spaink Herman P

    2011-06-01

    Full Text Available Abstract Background With the zebrafish increasingly being used in immunology and infectious disease research, there is a need for efficient molecular tools to evaluate immune gene expression in this model species. RT-MLPA (reverse transcription - multiplex ligation-dependent probe amplification provides a sensitive and reproducible method, in which fluorescently labelled amplification products of unique lengths are produced for a defined set of target transcripts. The method employs oligonucleotide probes that anneal to adjacent sites on a target sequence and are then joined by a heat-stable ligase. Subsequently, multiplex PCR with universal primers gives rise to amplicons that can be analyzed with standard sequencing equipment and relative quantification software. Allowing the simultaneous quantification of around 40 selected markers in a one-tube assay, RT-MLPA is highly useful for high-throughput screening applications. Findings We employed a dual-colour RT-MLPA probe design for chemical synthesis of probe pairs for 34 genes involved in Toll-like receptor signalling, transcriptional activation of the immune response, cytokine and chemokine production, and antimicrobial defence. In addition, six probe pairs were included for reference genes unaffected by infections in zebrafish. First, we established assay conditions for adult zebrafish infected with different strains of Mycobacterium marinum causing acute and chronic disease. Addition of competitor oligonucleotides was required to achieve peak heights in a similar range for genes with different expression levels. For subsequent analysis of embryonic samples it was necessary to adjust the amounts of competitor oligonucleotides, as the expression levels of several genes differed to a large extent between adult and embryonic tissues. Assay conditions established for one-day-old Salmonella typhimurium-infected embryos could be transferred without further adjustment to five-day-old M. marinum

  16. A Field-Tailored Reverse Transcription Loop-Mediated Isothermal Assay for High Sensitivity Detection of Plasmodium falciparum Infections

    Science.gov (United States)

    Kemleu, Sylvie; Guelig, Dylan; Eboumbou Moukoko, Carole; Essangui, Estelle; Diesburg, Steven; Mouliom, Abas; Melingui, Bernard; Manga, Jeanne; Donkeu, Christiane; Epote, Annie; Texier, Gaëtan; LaBarre, Paul; Burton, Robert

    2016-01-01

    Highly sensitive and field deployable molecular diagnostic tools are critically needed for detecting submicroscopic, yet transmissible levels of malaria parasites prevalent in malaria endemic countries worldwide. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed and evaluated in comparison with thick blood smear microscopy, an antigen-based rapid diagnostic test (RDT), and an in-house RT-PCR targeting the same RT-LAMP transcript. The optimized assay detected Plasmodium falciparum infections in as little as 0.25ng of total parasite RNA, and exhibited a detection limit of 0.08 parasites/ μL when tested directly on infected whole blood lysates, or ~0.0008 parasites/ μL when using RNA extracts. Assay positivity was observed as early as eight minutes from initiation of the RT-LAMP and in most cases the reaction was complete before twenty minutes. Clinical evaluation of the assay on 132 suspected malaria cases resulted in a positivity rate of 90% for RT-LAMP using extracted RNA, and 85% when using whole blood lysates. The positivity rates were 70% for P. falciparum-specific RDT, 83% for RT-PCR, and 74% for thick blood smear microscopy (Mean parasite density = 36,986 parasites/ μL). Concordance rates between the developed RT-LAMP and comparator tests were greater than 75%, the lowest being with light microscopy (78%, McNemar’s test: P = 0.0002), and the highest was with RT-PCR (87%, McNemar’s test: P = 0.0523). Compared to reference RT-PCR, assay sensitivity was 90% for RT-LAMP on whole blood, and 96% for RT-LAMP using corresponding RNA extracts. Electricity-free heaters were further developed and evaluated in comparison with a battery-operated isothermal amplification machine for use with the developed test in resource-limited settings. Taken together, the data highlight the benefits of targeting high abundant RNA transcripts in molecular diagnosis, as well as the potential usefulness of the developed RT-LAMP-assay in

  17. Reverse Transcription Polymerase Chain Reaction-based System for Simultaneous Detection of Multiple Lily-infecting Viruses

    Directory of Open Access Journals (Sweden)

    Ji Yeon Kwon

    2013-09-01

    Full Text Available A detection system based on a multiplex reverse transcription (RT polymerase chain reaction (PCR was developed to simultaneously identify multiple viruses in the lily plant. The most common viruses infecting lily plants are the cucumber mosaic virus (CMV, lily mottle virus (LMoV, lily symptomless virus (LSV. Leaf samples were collected at lily-cultivation facilities located in the Kangwon province of Korea and used to evaluate the detection system. Simplex and multiplex RT-PCR were performed using virus-specific primers to detect single-or mixed viral infections in lily plants. Our results demonstrate the selective detection of 3 different viruses (CMV, LMoV and LSV by using specific primers as well as the potential of simultaneously detecting 2 or 3 different viruses in lily plants with mixed infections. Three sets of primers for each target virus, and one set of internal control primers were used to evaluate the detection system for efficiency, reliability, and reproducibility.

  18. Strand-Specific Quantitative Reverse Transcription-Polymerase Chain Reaction Assay for Measurement of Arenavirus Genomic and Antigenomic RNAs.

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    Kelsey Haist

    Full Text Available Arenaviruses are bi-segmented, single-stranded RNA viruses that cause significant human disease. The manner in which they regulate the replication of their genome is not well-understood. This is partly due to the absence of a highly sensitive assay to measure individual species of arenavirus replicative RNAs. To overcome this obstacle, we designed a quantitative reverse transcription (RT-PCR assay for selective quantitation of each of the lymphocytic choriomeningitis virus (LCMV genomic or antigenomic RNAs. During the course of assay design, we identified a nonspecific priming phenomenon whereby, in the absence of an RT primer, cDNAs complementary to each of the LCMV replicative RNA species are generated during RT. We successfully circumvented this nonspecific priming event through the use of biotinylated primers in the RT reaction, which permitted affinity purification of primer-specific cDNAs using streptavidin-coated magnetic beads. As proof of principle, we used the assay to map the dynamics of LCMV replication at acute and persistent time points and to determine the quantities of genomic and antigenomic RNAs that are incorporated into LCMV particles. This assay can be adapted to measure total S or L segment-derived viral RNAs and therefore represents a highly sensitive diagnostic platform to screen for LCMV infection in rodent and human tissue samples and can also be used to quantify virus-cell attachment.

  19. Reliable gene expression analysis by reverse transcription-quantitative PCR: reporting and minimizing the uncertainty in data accuracy.

    Science.gov (United States)

    Remans, Tony; Keunen, Els; Bex, Geert Jan; Smeets, Karen; Vangronsveld, Jaco; Cuypers, Ann

    2014-10-01

    Reverse transcription-quantitative PCR (RT-qPCR) has been widely adopted to measure differences in mRNA levels; however, biological and technical variation strongly affects the accuracy of the reported differences. RT-qPCR specialists have warned that, unless researchers minimize this variability, they may report inaccurate differences and draw incorrect biological conclusions. The Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines describe procedures for conducting and reporting RT-qPCR experiments. The MIQE guidelines enable others to judge the reliability of reported results; however, a recent literature survey found low adherence to these guidelines. Additionally, even experiments that use appropriate procedures remain subject to individual variation that statistical methods cannot correct. For example, since ideal reference genes do not exist, the widely used method of normalizing RT-qPCR data to reference genes generates background noise that affects the accuracy of measured changes in mRNA levels. However, current RT-qPCR data reporting styles ignore this source of variation. In this commentary, we direct researchers to appropriate procedures, outline a method to present the remaining uncertainty in data accuracy, and propose an intuitive way to select reference genes to minimize uncertainty. Reporting the uncertainty in data accuracy also serves for quality assessment, enabling researchers and peer reviewers to confidently evaluate the reliability of gene expression data. © 2014 American Society of Plant Biologists. All rights reserved.

  20. Analysis of plasma viral RNA levels during acute dengue virus infection using quantitative competitor reverse transcription-polymerase chain reaction.

    Science.gov (United States)

    Sudiro, T M; Zivny, J; Ishiko, H; Green, S; Vaughn, D W; Kalayanarooj, S; Nisalak, A; Norman, J E; Ennis, F A; Rothman, A L

    2001-01-01

    There is increasing recognition of the potential importance of viral burden in the pathogenesis of dengue hemorrhagic fever (DHF). There is little data available, however, describing the kinetics of viral replication in humans with natural dengue virus (DV) infection. Standard procedures for measuring titers of infectious virus in clinical specimens are either laborious or insensitive. We developed a method for measurement of DV RNA in plasma samples based on reverse transcription-polymerase chain reaction (RT-PCR) using a mutant RNA target as a competitor. This technique was reproducible and accurate for samples containing any of the four DV serotypes, and could be applied to samples containing as few as 250 copies of RNA per reaction. We examined plasma viral RNA levels in 80 children with acute DV infection; sequential plasma samples were tested in 34 of these children. Plasma viral RNA levels ranged as high as 10(9) RNA copies/ml, and correlated with titers of infectious virus measured in mosquitoes (r= 0.69). Plasma viral RNA levels fell rapidly during the last several days of the febrile period. We did not find a significant difference in maximal plasma viral RNA levels between children with DHF and children with dengue fever, but peak viral RNA levels were identified in only 16 subjects. We conclude that this quantitative RT-PCR method will be valuable for further studies of natural DV infections.

  1. Development and application of reverse transcription loop-mediated isothermal amplification for detecting live Shewanella putrefaciens in preserved fish sample.

    Science.gov (United States)

    Li, Chenghua; Ying, Qi; Su, Xiurong; Li, Taiwu

    2012-04-01

    Given that live Shewanella putrefaciens is one of the major causes of spoilage for aquatic products even in chill storage, the rapid and accurate detection process is the first priority. In the present study, a novel reverse transcription loop-mediated isothermal amplification (RT-LAMP) detecting assay was developed by targeting internal transcribed spacer (ITS) sequence between 16S and 23S rRNA. At the same time, a new procaryotic mRNA isolation strategy was also established by introducing a polyA tail to RNA during cDNA synthesis step. Under the optimal reaction time (60 min) and temperature (64.1 °C), S. putrefaciens could be specially identified from a variety of other tested bacteria by RT-LAMP. The sensitivity analysis showed that RT-LAMP could be identified as lower as 5.4 copies per reaction, which is over 200-fold higher than that of standard PCR (1.08 × 10³ copies per reaction). The method could be effectively identified S. putrefaciens in artificially contaminated or spoilaged fish samples with dose-dependent manners. To our knowledge, this is the first report using RT-LAMP assay to detect live S. putrefaciens in fish. The study provided a rapid and accurate detection method for live bacteria in aquatic food and established a new procaryotic mRNA isolation strategy at the same time, which will be useful for food preservation. © 2012 Institute of Food Technologists®

  2. Identification of viable Listeria species based on reverse transcription-multiplex PCR (RT-MPCR) and restriction digestion.

    Science.gov (United States)

    Rattanachaikunsopon, Pongsak; Phumkhachorn, Parichat

    2012-01-01

    A novel method for the identification of viable Listeria species was developed based on reverse transcription-multiplex PCR (RT-MPCR) and restriction digestion. The targets for RT-MPCR were iap mRNAs whose genes are common to all Liseria species. A set of five primers was used in this study. Two of them were genus specific, and the other three were specific to L. monocytogenase, L. innocua, and L. grayi respectively. By RT-MPCR, L. monocytogenese, L. innocua, L. grayi, and a group of Listeria species, including L. ivanovii, L. welshimeri, and L. seeligeri, were specifically identified. To differentiate the latter three Listeria species, RT-MPCR products were subjected to digestion with HpaI and ScaI. The sensitivity of RT-MPCR in detecting Listeria species was determined to be 50 CFU/mL. RT-MPCR was found to discriminate between viable and nonviable cells and to detect viable Listeria species in a food model.

  3. Detection of Avian bornavirus in multiple tissues of infected psittacine birds using real-time reverse transcription polymerase chain reaction.

    Science.gov (United States)

    Delnatte, Pauline; Mak, Matthew; Ojkic, Davor; Raghav, Raj; DeLay, Josepha; Smith, Dale A

    2014-03-01

    Avian bornavirus (ABV), the cause of proventricular dilation disease in psittacine birds, has been detected in multiple tissues of infected birds using immunohistochemical staining (IHC) and reverse transcription polymerase chain reaction (RT-PCR). In the current study, real-time RT-PCR, using primers targeting the ABV matrix gene, was used to detect ABV in 146 tissues from 7 ABV-infected psittacine birds. Eighty-six percent of the samples tested positive, with crossing point values ranging from 13.82 to 37.82 and a mean of 22.3. These results were compared to the findings of a previous study using gel-based RT-PCR and IHC on the same samples. The agreement between the 2 RT-PCR techniques was 91%; when tests disagreed it was because samples were negative using gel-based RT-PCR but positive on real-time RT-PCR. Agreement with IHC was 77%; 16 out of 74 samples were negative using IHC but positive on real-time RT-PCR. The results suggest that real-time RT-PCR is a more sensitive technique than gel-based RT-PCR and IHC to detect ABV in tissues. The tissues that were ranked most frequently as having a high amount of viral RNA were proventriculus, kidney, colon, cerebrum, and cerebellum. Skeletal muscle, on the other hand, was found to have a consistently low amount of viral RNA.

  4. Candidate gene biodosimeters of mice and human exposure to ionizing radiation by quantitative reverse transcription polymerase chain reaction

    Directory of Open Access Journals (Sweden)

    Hamed Rezaeejam

    2015-01-01

    Full Text Available Understanding of cellular responses to ionizing radiation (IR is essential for the development of predictive markers useful for assessing human exposure. Biological markers of exposure to IR in human populations are of great interest for assessing normal tissue injury in radiation oncology and for biodosimetry in nuclear incidents and accidental radiation exposures. Traditional radiation exposure biomarkers based on cytogenetic assays (biodosimetry, are time-consuming and do not provide results fast enough and requires highly trained personnel for scoring. Hence, the development of rapid biodosimetry methods is one of the highest priorities. Exposure of cells to IR activates multiple signal transduction pathways, which result in complex alterations in gene-expression. Real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR has become the benchmark for the detection and quantification of RNA targets and is being utilized increasingly in monitoring the specific genes with more accurately and sensitively. This review evaluates the RT-qPCR as a biodosimetry method and we investigated the papers from 2000 up to now, which identified the genes-expression related the DNA repair, cell cycle checkpoint, and apoptosis induced by ionization radiation in peripheral blood and determined as biodosimeters. In conclusion, it could be say that RT-qPCR technique for determining the specific genes as biodosimeters could be a fully quantitative reliable and sensitive method. Furthermore, the results of the current review will help the researchers to recognize the most expressed genes induced by ionization radiation.

  5. RT-Bst: an integrated approach for reverse transcription and enrichment of cDNA from viral RNA.

    Science.gov (United States)

    Kabir, M S; Clements, M O; Kimmitt, P T

    2015-01-01

    The synthesis of cDNA from RNA is challenging due to the inefficiency of reverse transcription (RT). In order to address this, an RT-Bst method was developed for sequential RT of RNA and Bst DNA polymerase amplification for enrichment of cDNA in a single-tube reaction. Using genomic RNA from bacteriophage MS2, the yield of cDNA produced by RT alone and RT-Bst were compared by analysis of polymerase chain reaction (PCR)-amplified products. A superior performance was observed when amplifying MS2 cDNA with random primers following RT-Bst compared to RT alone, indicating greater quantities of cDNA were present after RT-Bst. RT-Bst was also compared with RT alone for their relative ability to produce sufficient cDNA to amplify eight target regions spanning the respiratory syncytial virus (RSV) genome. Six out of eight targets were amplified consistently by PCR subsequent to RT-Bst amplification, whereas only three out of eight targets could be amplified after RT alone. The RSV sequences were selectively amplified using RSV-specific primers from a mixed template containing an excess of MS2 RNA without amplifying MS2 sequences. This suggests that RT-Bst can be used to amplify RNA sequences non-specifically using random primers and specifically using sequence-specific primers, and enhances the yield of cDNA when compared to RT alone.

  6. Real-Time Quantitative PCR (QPCR) and Reverse Transcription-QPCR for Detection and Enumeration of Total Yeasts in Wine▿

    Science.gov (United States)

    Hierro, Núria; Esteve-Zarzoso, Braulio; González, Ángel; Mas, Albert; Guillamón, Jose M.

    2006-01-01

    Real-time PCR, or quantitative PCR (QPCR), has been developed to rapidly detect and quantify the total number of yeasts in wine without culturing. Universal yeast primers were designed from the variable D1/D2 domains of the 26S rRNA gene. These primers showed good specificity with all the wine yeasts tested, and they did not amplify the most representative wine species of acetic acid bacteria and lactic acid bacteria. Numerous standard curves were constructed with different strains and species grown in yeast extract-peptone-dextrose medium or incubated in wine. The small standard errors with these replicas proved that the assay is reproducible and highly robust. This technique was validated with artificially contaminated and natural wine samples. We also performed a reverse transcription-QPCR (RT-QPCR) assay from rRNA for total viable yeast quantification. This technique had a low detection limit and was more accurate than QPCR because the dead cells were not quantified. As far as we know, this is the first time that RT-QPCR has been performed to quantify viable yeasts from rRNA. RT-QPCR is a rapid and accurate technique for enumerating yeasts during industrial wine fermentation and controlling the risk of wine spoilage. PMID:17088381

  7. A multiplex reverse transcription-polymerase chain reaction assay for Newcastle disease virus and avian pneumovirus (Colorado strain).

    Science.gov (United States)

    Ali, A; Reynolds, D L

    2000-01-01

    Newcastle disease virus (NDV) and avian pneumovirus (APV) cause Newcastle disease and rhinotracheitis respectively, in turkeys. Both of these viruses infect the respiratory system. A one-tube, multiplex, reverse transcription-polymerase chain reaction (RT-PCR) assay for the detection of both NDV and Colorado strain of APV (APV-Col) was developed and evaluated. The primers, specific for each virus, were designed from the matrix protein gene of APV-Col and the fusion protein gene of NDV to amplify products of 631 and 309 nucleotides, respectively. The multiplex RT-PCR assay, for detecting both viruses simultaneously, was compared with the single-virus RT-PCR assays for its sensitivity and specificity. The specific primers amplified products of predicted size from each virus in the multiplex as well as the single-virus RT-PCR assays. The multiplex RT-PCR assay was determined to be equivalent to the single-virus RT-PCR assays for detecting both NDV and APV-Col. This multiplex RT-PCR assay proved to be a sensitive method for the simultaneous and rapid detection of NDV and APV-Col. This assay has the potential for clinical diagnostic applications.

  8. Comparison between Saliva and Nasopharyngeal Swab Specimens for Detection of Respiratory Viruses by Multiplex Reverse Transcription-PCR.

    Science.gov (United States)

    Kim, Young-Gon; Yun, Seung Gyu; Kim, Min Young; Park, Kwisung; Cho, Chi Hyun; Yoon, Soo Young; Nam, Myung Hyun; Lee, Chang Kyu; Cho, Yun-Jung; Lim, Chae Seung

    2017-01-01

    Nasopharyngeal swabs (NPSs) are being widely used as specimens for multiplex real-time reverse transcription (RT)-PCR for respiratory virus detection. However, it remains unclear whether NPS specimens are optimal for all viruses targeted by multiplex RT-PCR. In addition, the procedure to obtain NPS specimens causes coughing in most patients, which possibly increases the risk of nosocomial spread of viruses. In this study, paired NPS and saliva specimens were collected from 236 adult male patients with suspected acute respiratory illnesses. Specimens were tested for 16 respiratory viruses by multiplex real-time RT-PCR. Among the specimens collected from the 236 patients, at least 1 respiratory virus was detected in 183 NPS specimens (77.5%) and 180 saliva specimens (76.3%). The rates of detection of respiratory viruses were comparable for NPS and saliva specimens (P = 0.766). Nine virus species and 349 viruses were isolated, 256 from NPS specimens and 273 from saliva specimens (P = 0.1574). Adenovirus was detected more frequently in saliva samples (P saliva samples was excluded by direct sequencing. In conclusion, neither of the sampling methods was consistently more sensitive than the other. We suggest that these cost-effective methods for detecting respiratory viruses in mixed NPS-saliva specimens might be valuable for future studies. Copyright © 2016 American Society for Microbiology.

  9. One-Step Generation of Scalable Multiparticle Entanglement for Hot Ions Driven by a Standing-Wave Laser

    Institute of Scientific and Technical Information of China (English)

    杨文星; 陈爱喜

    2011-01-01

    An alternative scheme is proposed for one-step generation of multiparticle cluster state with trapped ions in thermal motion. In this scheme, the ions are simultaneously illuminated by a standing-wave laser tuned to the carrier. During the operations, the vibrational mode is virtually excited, thus the quantum operations are insensitive to the heating. It is shown that the high fidelity multiparticle entanglement could be generated in just one step even including the small fluctuations of parameters. In addition, the ion does not need to be exactly positioned at the node of the standing wave, which is also important from the viewpoint of experiment.

  10. Synthesis of Carbon Nanotube/Graphene Composites by One-Step Chemical Vapor Deposition for Electrodes of Electrochemical Capacitors

    Directory of Open Access Journals (Sweden)

    Chuen-Chang Lin

    2015-01-01

    Full Text Available To control the packing density of carbon nanotubes (CNTs and the number of graphene layers, carbon nanotube/graphene composites are directly grown on cobalt (Co catalysts-coated nickel foam by one-step ambient pressure chemical vapor deposition (CVD at different temperatures and times. The carbon nanotube/graphene composites grown by one-step CVD at 850°C for 10 min possess the highest specific capacitance. Furthermore, a lower growing temperature leads to a higher packing density of CNTs and a smaller number of layers of graphene. A shorter growing time also leads to a smaller number of layers of graphene.

  11. Rapid and sensitive electrochemiluminescence detection of rotavirus by magnetic primer based reverse transcription-polymerase chain reaction

    Energy Technology Data Exchange (ETDEWEB)

    Zhan Fangfang; Zhou Xiaoming [MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631 (China); Xing Da, E-mail: xingda@scnu.edu.cn [MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631 (China)

    2013-01-25

    Graphical abstract: In this work, we have developed and demonstrated a magnetic primer based RT-PCR assay for ECL detection of rotavirus. In the presence of two functional primers (magnetic primer and TBR-primer) and PCR reagents, cDNA from RT was amplified directly onto MPs during PCR cycles of denaturation, annealing and extension. The resulting MPs-TBR complexes were easily loaded on the electrode surface and produced a concentrated ECL signal. The figure shows the schematic illustration of magnetic primer RT-PCR based ECL assay for rotavirus detection. Highlights: Black-Right-Pointing-Pointer A novel method for detection of rotavirus has been developed. Black-Right-Pointing-Pointer In the presence of magnetic primer, TBR-primer and PCR reagents, cDNA form RT was amplified directly onto MPs. Black-Right-Pointing-Pointer To obtain the best sensing and efficient performance, important parameters associated with the efficiency were investigated carefully. Black-Right-Pointing-Pointer The proposed method will find numerous applications in food safety field and clinical diagnosis. - Abstract: A novel method for detection of rotavirus has been developed by integrating magnetic primer based reverse transcription-polymerase chain reaction (RT-PCR) with electrochemiluminescence (ECL) detection. This is realized by accomplishing RT of rotavirus RNA in traditional way and performing PCR of the resulting cDNA fragment on the surface of magnetic particles (MPs). In order to implement PCR on MPs and achieve rapid ECL detection, forward and reverse primers are bounded to MPs and tris-(2,2 Prime -bipyridyl) ruthenium (TBR), respectively. After RT-PCR amplification, the TBR labels are directly enriched onto the surface of MPs. Then the MPs-TBR complexes can be loaded on the electrode surface and analyzed by magnetic ECL platform without any post-modification or post-incubation process. So some laborious manual operations can be avoided to achieve rapid yet sensitive detection

  12. Inhibition of both HIV-1 reverse transcription and gene expression by a cyclic peptide that binds the Tat-transactivating response element (TAR RNA.

    Directory of Open Access Journals (Sweden)

    Matthew S Lalonde

    2011-05-01

    Full Text Available The RNA response element TAR plays a critical role in HIV replication by providing a binding site for the recruitment of the viral transactivator protein Tat. Using a structure-guided approach, we have developed a series of conformationally-constrained cyclic peptides that act as structural mimics of the Tat RNA binding region and block Tat-TAR interactions at nanomolar concentrations in vitro. Here we show that these compounds block Tat-dependent transcription in cell-free systems and in cell-based reporter assays. The compounds are also cell permeable, have low toxicity, and inhibit replication of diverse HIV-1 strains, including both CXCR4-tropic and CCR5-tropic primary HIV-1 isolates of the divergent subtypes A, B, C, D and CRF01_AE. In human peripheral blood mononuclear cells, the cyclic peptidomimetic L50 exhibited an IC(50 ∼250 nM. Surprisingly, inhibition of LTR-driven HIV-1 transcription could not account for the full antiviral activity. Timed drug-addition experiments revealed that L-50 has a bi-phasic inhibition curve with the first phase occurring after HIV-1 entry into the host cell and during the initiation of HIV-1 reverse transcription. The second phase coincides with inhibition of HIV-1 transcription. Reconstituted reverse transcription assays confirm that HIV-1 (- strand strong stop DNA synthesis is blocked by L50-TAR RNA interactions in-vitro. These findings are consistent with genetic evidence that TAR plays critical roles both during reverse transcription and during HIV gene expression. Our results suggest that antiviral drugs targeting TAR RNA might be highly effective due to a dual inhibitory mechanism.

  13. A one-step molecular biology method for simple and rapid detection of grass carp Ctenopharyngodon idella reovirus (GCRV) HZ08 strain

    Science.gov (United States)

    Six reverse-transcription loop-mediated isothermal amplification (RT-LAMP) primers designed against conserved regions of segment 6 (s6) gene were used for the detection of grass carp Ctenopharyngodon idella reovirus (GCRV) HZ08 subtype. The entire amplification could be completed within 40 min at 62...

  14. One-step electrochemical growth of a three-dimensional Sn-Ni@PEO nanotube array as a high performance lithium-ion battery anode.

    Science.gov (United States)

    Fan, Xin; Dou, Peng; Jiang, Anni; Ma, Daqian; Xu, Xinhua

    2014-12-24

    Various well-designed nanostructures have been proposed to optimize the electrode systems of lithium-ion batteries for problems like Li(+) diffusion, electron transport, and large volume changes so as to fulfill effective capacity utilization and increase electrode stability. Here, a novel three-dimensional (3D) hybrid Sn-Ni@PEO nanotube array is synthesized as a high performance anode for a lithium-ion battery through a simple one-step electrodeposition for the first time. Superior to the traditional stepwise synthesis processes of heterostructured nanomaterials, this one-step method is more suitable for practical applications. The electrode morphology is well preserved after repeated Li(+) insertion and extraction, indicating that the positive synergistic effect of the alloy nanotube array and 3D ultrathin PEO coating could authentically optimize the current volume-expansion electrode system. The electrochemistry results further confirm that the superiority of the Sn-Ni@PEO nanotube array electrode could largely boost durable high reversible capacities and superior rate performances compared to a Sn-Ni nanowire array. This proposed ternary hybrid structure is proven to be an ideal candidate for the development of high performance anodes for lithium-ion batteries.

  15. DETECTION OF MICROMETASTASES OF LUNG CANCER BY USING LUNX mRNA SPECIFIC REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION

    Institute of Scientific and Technical Information of China (English)

    朱广迎; 刘德林; 王绪; 彭猛青; 刘惠; 沈万华; 张海舟; 王伟; 陈杰

    2002-01-01

    Objective: To detect of lung cancer micrometastases in peripheral blood and regional lymphatic nodes by using lunx mRNA specific reverse transcription-polymerase chain reaction (RT-PCR). Methods: RT-PCR was used to detect lunx mRNA in peripheral blood of 26 patients with lung cancer. We also detected 44 regional lymphatic nodes obtained from 25 patients with lung cancer who underwent curative lobectomy. All the 44 regional lymphatic nodes were also examined by histopathology. Micrometastatic tumor cells in the peripheral blood and regional lymphatic nodes were semiquantitatively determined with the ratio of lunx band intensity to the glyceraldehydes-3-phosphate dehydrogenase band intensity. Results: The positive detection rate of lunx mRNA in peripheral blood for non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC) patients were 60% (12/20) and 67% (4/6) respectively. 16 (36.4%) of regional lymphatic nodes from 44 lung cancer patients were positive by RT-PCR while 6 (13.6%) were positive by histopathology (x2=6.06, P=0.014). However, no blood samples and lymphatic nodes from patients with benign pulmonary diseases or normal volunteers were positive for lunx mRNA. The positive detection rate of lunx mRNA in bone marrow of NSCLC amd SCLC patients were 65% (13/20) and 67% (4/6) respectively. Conclusion: RT-PCR amplification of lunx mRNA is an sensitive and specific means to detect early haematogenous and regional lymphatic nodes dissemination of cancer cells for patients with lung cancer.

  16. Rapid detection of all known ebolavirus species by reverse transcription-loop-mediated isothermal amplification (RT-LAMP).

    Science.gov (United States)

    Oloniniyi, Olamide K; Kurosaki, Yohei; Miyamoto, Hiroko; Takada, Ayato; Yasuda, Jiro

    2017-03-26

    Ebola virus disease (EVD), a highly virulent infectious disease caused by ebolaviruses, has a fatality rate of 25-90%. Without a licensed chemotherapeutic agent or vaccine for the treatment and prevention of EVD, control of outbreaks requires accurate and rapid diagnosis of cases. In this study, five sets of six oligonucleotide primers targeting the nucleoprotein gene were designed for specific identification of each of the five ebolavirus species using reverse transcription-loop mediated isothermal amplification (RT-LAMP) assay. The detection limits of the ebolavirus species-specific primer sets were evaluated using in vitro transcribed RNAs. The detection limit of species-specific RT-LAMP assays for Zaire ebolavirus, Sudan ebolavirus, Taï Forest ebolavirus, and Bundibugyo ebolavirus was 256 copies/reaction, while the detection limit for Reston ebolavirus was 64 copies/reaction, and the detection time for each of the RT-LAMP assays was 13.3±3.0, 19.8±4.6, 14.3±0.6, 16.1±4.7, and 19.8±2.4min (mean±SD), respectively. The sensitivity of the species-specific RT-LAMP assays were similar to that of the established RT-PCR and quantitative RT-PCR assays for diagnosis of EVD and are suitable for field or point-of-care diagnosis. The RT-LAMP assays were specific for the detection of the respective species of ebolavirus with no cross reaction with other species of ebolavirus and other viral hemorrhagic fever viruses such as Marburg virus, Lassa fever virus, and Dengue virus. The species-specific RT-LAMP assays developed in this study are rapid, sensitive, and specific and could be useful in case of an EVD outbreak.

  17. One-Pot Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP) for Detecting MERS-CoV.

    Science.gov (United States)

    Lee, Se Hee; Baek, Yun Hee; Kim, Yang-Hoon; Choi, Young-Ki; Song, Min-Suk; Ahn, Ji-Young

    2016-01-01

    Due to the limitation of rapid development of specific antiviral drug or vaccine for novel emerging viruses, an accurate and rapid diagnosis is a key to manage the virus spread. We developed an efficient and rapid method with high specificity for the Middle East Respiratory Syndrome coronavirus (MERS-CoV), based on one-pot reverse transcription loop-mediated isothermal amplification (one-pot RT-LAMP). A set of six LAMP primers [F3, B3, FIP, BIP, LF (Loop-F), and LB (Loop-B)] were designed using the sequence of nucleocapsid (N) gene with optimized RT-LAMP enzyme conditions: 100 U M-MLV RTase and 4 U Bst polymerase, implying that the reaction was able to detect four infectious viral genome copies of MERS-CoV within a 60 min reaction time period. Significantly, EvaGreen dye has better signal read-out properties in one-pot RT-LAMP reaction and is more compatible with DNA polymerase than SYBR green I. Isothermally amplified specific N genes were further evaluated using field-deployable microchamber devices, leading to the specific identification of as few as 0.4 infectious viral genome copies, with no cross-reaction to the other acute respiratory disease viruses, including influenza type A (H1N1 and H3N2), type B, human coronavirus 229E, and human metapneumovirus. This sensitive, specific and feasible method provides a large-scale technical support in emergencies, and is also applied as a sample-to-detection module in Point of Care Testing devices.

  18. Improved detection of Lassa virus by reverse transcription-PCR targeting the 5' region of S RNA.

    Science.gov (United States)

    Olschläger, Stephan; Lelke, Michaela; Emmerich, Petra; Panning, Marcus; Drosten, Christian; Hass, Meike; Asogun, Danny; Ehichioya, Deborah; Omilabu, Sunday; Günther, Stephan

    2010-06-01

    The method of choice for the detection of Lassa virus is reverse transcription (RT)-PCR. However, the high degree of genetic variability of the virus poses a problem with the design of RT-PCR assays that will reliably detect all strains. Recently, we encountered difficulties in detecting some strains from Liberia and Nigeria in a commonly used glycoprotein precursor (GPC) gene-specific RT-PCR assay (A. H. Demby, J. Chamberlain, D. W. Brown, and C. S. Clegg, J. Clin. Microbiol. 32:2898-2903, 1994), which prompted us to revise the protocol. The design of the new assay, the GPC RT-PCR/2007 assay, took into account 62 S RNA sequences from all countries where Lassa fever is endemic, including 40 sequences generated from the strains in our collection. The analytical sensitivity of the new assay was determined with 11 strains from Sierra Leone, Liberia, Ivory Coast, and Nigeria by probit analysis; the viral loads detectable with a probability of 95% ranged from 342 to 2,560 S RNA copies/ml serum, which corresponds to 4 to 30 S RNA copies/assay. The GPC RT-PCR/2007 assay was validated with 77 serum samples and 1 cerebrospinal fluid sample from patients with laboratory-confirmed Lassa fever. The samples mainly originated from Liberia and Nigeria and included strains difficult to detect in the assay of 1994. The GPC RT-PCR/2007 assay detected virus in all clinical specimens (100% sensitivity). In conclusion, a new RT-PCR assay, based in part on the protocol developed by Demby et al. in 1994, for the detection of Lassa virus is described. Compared to the assay developed in 1994, the GPC RT-PCR/2007 assay offers improved sensitivity for the detection of Liberian and Nigerian Lassa virus strains.

  19. Evaluation of Altona Diagnostics RealStar Zika Virus Reverse Transcription-PCR Test Kit for Zika Virus PCR Testing.

    Science.gov (United States)

    L'Huillier, Arnaud G; Lombos, Ernesto; Tang, Elaine; Perusini, Stephen; Eshaghi, Alireza; Nagra, Sandeep; Frantz, Christine; Olsha, Romy; Kristjanson, Erik; Dimitrova, Kristina; Safronetz, David; Drebot, Mike; Gubbay, Jonathan B

    2017-05-01

    With the emerging Zika virus (ZIKV) epidemic, accessible real-time reverse transcription-PCR (rRT-PCR) assays are needed to streamline testing. The commercial Altona Diagnostics RealStar ZIKV rRT-PCR test kit (Altona PCR) has been approved for emergency use authorization by the U.S. FDA. Our aim was to verify the Altona PCR by comparing it to the CDC-designed dual-target ZIKV rRT-PCR reference assay (reference PCR) and describe the demographics of patients tested for ZIKV by rRT-PCR in Ontario, Canada. A large set of clinical specimens was tested for ZIKV by the Altona PCR and the reference PCR. Positive or equivocal specimens underwent PCR and Sanger sequencing targeting the ZIKV NS5 gene. A total of 671 serum specimens were tested by the reference PCR: 58 (8.6%) were positive, 193 (28.8%) were equivocal, and 420 (62.6%) were negative. Ninety percent of the reference PCR-positive patients were tested in the first 5 days after symptom onset. The Altona PCR was performed on 284/671 specimens tested by the reference PCR. The Altona PCR was positive for 53/58 (91%) reference PCR-positive specimens and 16/193 (8%) reference PCR-equivocal specimens; the ZIKV NS5 PCR was positive for all 68 Altona PCR-positive specimens and negative for all 181 Altona PCR-negative specimens that underwent the NS5 PCR. The Altona PCR has very good sensitivity (91%) and specificity (97%) compared to the reference PCR. The Altona PCR can be used for ZIKV diagnostic testing and has less extensive verification requirements than a laboratory-developed test. Copyright © 2017 American Society for Microbiology.

  20. Aphids preserved in propylene glycol can be used for reverse transcription-polymerase chain reaction detection of Potato virus Y.

    Science.gov (United States)

    Nie, Xianzhou; Pelletier, Yvan; Mason, Nicola; Dilworth, Andrea; Giguère, Marie-Andrée

    2011-08-01

    The effectiveness of propylene glycol on the retention of RNA target of Potato virus Y (PVY), an aphid stylet-borne virus, in Myzus persicae was investigated in comparison to ethanol and liquid nitrogen/-80°C. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the PVY targets from the propylene glycol/ethanol/liquid nitrogen preserved single aphids after a 5min acquisition period from infected potato plants. In the liquid nitrogen/-80°C and 70% ethanol treatments, 55.6% and 38.8% aphids tested PVY-positive, respectively. In the 0-75% propylene glycol treatments, 12.2-44.7% aphids tested PVY-positive. The lowest detection rate was in the 0% (positive rate, 15.2%) and the 10% propylene glycol (positive rate, 12.2%). As the propylene glycol concentration increased to 25%, 29.8% aphids tested positive. A high PVY-positive rate was also found in 35-75% propylene glycol treatments at 44.7% (35% propylene glycol), 36.7% (50% propylene glycol) and 34.8% (75% propylene glycol), which is comparable to the rate shown in 70% ethanol. No significant difference in the positive detection rate was observed in aphids preserved in 50% propylene glycol at room temperature for 2, 4 and 10 days. These results demonstrate that propylene glycol at 25-75% can retain PVY targets effectively in aphids for an extended time period, and thus can be used in aphid traps to preserve viruliferous aphids for later RT-PCR detection of PVY.

  1. Intracerebral Borna disease virus infection of bank voles leading to peripheral spread and reverse transcription of viral RNA.

    Directory of Open Access Journals (Sweden)

    Paula Maria Kinnunen

    Full Text Available Bornaviruses, which chronically infect many species, can cause severe neurological diseases in some animal species; their association with human neuropsychiatric disorders is, however, debatable. The epidemiology of Borna disease virus (BDV, as for other members of the family Bornaviridae, is largely unknown, although evidence exists for a reservoir in small mammals, for example bank voles (Myodes glareolus. In addition to the current exogenous infections and despite the fact that bornaviruses have an RNA genome, bornavirus sequences integrated into the genomes of several vertebrates millions of years ago. Our hypothesis is that the bank vole, a common wild rodent species in traditional BDV-endemic areas, can serve as a viral host; we therefore explored whether this species can be infected with BDV, and if so, how the virus spreads and whether viral RNA is transcribed into DNA in vivo.We infected neonate bank voles intracerebrally with BDV and euthanized them 2 to 8 weeks post-infection. Specific Ig antibodies were detectable in 41%. Histological evaluation revealed no significant pathological alterations, but BDV RNA and antigen were detectable in all infected brains. Immunohistology demonstrated centrifugal spread throughout the nervous tissue, because viral antigen was widespread in peripheral nerves and ganglia, including the mediastinum, esophagus, and urinary bladder. This was associated with viral shedding in feces, of which 54% were BDV RNA-positive, and urine at 17%. BDV nucleocapsid gene DNA occurred in 66% of the infected voles, and, surprisingly, occasionally also phosphoprotein DNA. Thus, intracerebral BDV infection of bank vole led to systemic infection of the nervous tissue and viral excretion, as well as frequent reverse transcription of the BDV genome, enabling genomic integration. This first experimental bornavirus infection in wild mammals confirms the recent findings regarding bornavirus DNA, and suggests that bank voles are

  2. Intracerebral Borna disease virus infection of bank voles leading to peripheral spread and reverse transcription of viral RNA.

    Science.gov (United States)

    Kinnunen, Paula Maria; Inkeroinen, Hanna; Ilander, Mette; Kallio, Eva Riikka; Heikkilä, Henna Pauliina; Koskela, Esa; Mappes, Tapio; Palva, Airi; Vaheri, Antti; Kipar, Anja; Vapalahti, Olli

    2011-01-01

    Bornaviruses, which chronically infect many species, can cause severe neurological diseases in some animal species; their association with human neuropsychiatric disorders is, however, debatable. The epidemiology of Borna disease virus (BDV), as for other members of the family Bornaviridae, is largely unknown, although evidence exists for a reservoir in small mammals, for example bank voles (Myodes glareolus). In addition to the current exogenous infections and despite the fact that bornaviruses have an RNA genome, bornavirus sequences integrated into the genomes of several vertebrates millions of years ago. Our hypothesis is that the bank vole, a common wild rodent species in traditional BDV-endemic areas, can serve as a viral host; we therefore explored whether this species can be infected with BDV, and if so, how the virus spreads and whether viral RNA is transcribed into DNA in vivo.We infected neonate bank voles intracerebrally with BDV and euthanized them 2 to 8 weeks post-infection. Specific Ig antibodies were detectable in 41%. Histological evaluation revealed no significant pathological alterations, but BDV RNA and antigen were detectable in all infected brains. Immunohistology demonstrated centrifugal spread throughout the nervous tissue, because viral antigen was widespread in peripheral nerves and ganglia, including the mediastinum, esophagus, and urinary bladder. This was associated with viral shedding in feces, of which 54% were BDV RNA-positive, and urine at 17%. BDV nucleocapsid gene DNA occurred in 66% of the infected voles, and, surprisingly, occasionally also phosphoprotein DNA. Thus, intracerebral BDV infection of bank vole led to systemic infection of the nervous tissue and viral excretion, as well as frequent reverse transcription of the BDV genome, enabling genomic integration. This first experimental bornavirus infection in wild mammals confirms the recent findings regarding bornavirus DNA, and suggests that bank voles are capable of

  3. Development and validation of a multiplex reverse transcription PCR assay for simultaneous detection of three papaya viruses.

    Science.gov (United States)

    Tuo, Decai; Shen, Wentao; Yang, Yong; Yan, Pu; Li, Xiaoying; Zhou, Peng

    2014-10-21

    Papaya ringspot virus (PRSV), Papaya leaf distortion mosaic virus (PLDMV), and Papaya mosaic virus (PapMV) produce similar symptoms in papaya. Each threatens commercial production of papaya on Hainan Island, China. In this study, a multiplex reverse transcription PCR assay was developed to detect simultaneously these three viruses by screening combinations of mixed primer pairs and optimizing the multiplex RT-PCR reaction conditions. A mixture of three specific primer pairs was used to amplify three distinct fragments of 613 bp from the P3 gene of PRSV, 355 bp from the CP gene of PLDMV, and 205 bp from the CP gene of PapMV, demonstrating the assay's specificity. The sensitivity of the multiplex RT-PCR was evaluated by showing plasmids containing each of the viral target genes with 1.44 × 103, 1.79 × 103, and 1.91 × 102 copies for the three viruses could be detected successfully. The multiplex RT-PCR was applied successfully for detection of three viruses from 341 field samples collected from 18 counties of Hainan Island, China. Rates of single infections were 186/341 (54.5%), 93/341 (27.3%), and 3/341 (0.9%), for PRSV, PLDMV, and PapMV, respectively; 59/341 (17.3%) of the samples were co-infected with PRSV and PLDMV, which is the first time being reported in Hainan Island. This multiplex RT-PCR assay is a simple, rapid, sensitive, and cost-effective method for detecting multiple viruses in papaya and can be used for routine molecular diagnosis and epidemiological studies in papaya.

  4. Development and Validation of a Multiplex Reverse Transcription PCR Assay for Simultaneous Detection of Three Papaya Viruses

    Directory of Open Access Journals (Sweden)

    Decai Tuo

    2014-10-01

    Full Text Available Papaya ringspot virus (PRSV, Papaya leaf distortion mosaic virus (PLDMV, and Papaya mosaic virus (PapMV produce similar symptoms in papaya. Each threatens commercial production of papaya on Hainan Island, China. In this study, a multiplex reverse transcription PCR assay was developed to detect simultaneously these three viruses by screening combinations of mixed primer pairs and optimizing the multiplex RT-PCR reaction conditions. A mixture of three specific primer pairs was used to amplify three distinct fragments of 613 bp from the P3 gene of PRSV, 355 bp from the CP gene of PLDMV, and 205 bp from the CP gene of PapMV, demonstrating the assay’s specificity. The sensitivity of the multiplex RT-PCR was evaluated by showing plasmids containing each of the viral target genes with 1.44 × 103, 1.79 × 103, and 1.91 × 102 copies for the three viruses could be detected successfully. The multiplex RT-PCR was applied successfully for detection of three viruses from 341 field samples collected from 18 counties of Hainan Island, China. Rates of single infections were 186/341 (54.5%, 93/341 (27.3%, and 3/341 (0.9%, for PRSV, PLDMV, and PapMV, respectively; 59/341 (17.3% of the samples were co-infected with PRSV and PLDMV, which is the first time being reported in Hainan Island. This multiplex RT-PCR assay is a simple, rapid, sensitive, and cost-effective method for detecting multiple viruses in papaya and can be used for routine molecular diagnosis and epidemiological studies in papaya.

  5. Selection of reference genes for quantitative reverse-transcription polymerase chain reaction normalization in Brassica napus under various stress conditions.

    Science.gov (United States)

    Wang, Zheng; Chen, Yu; Fang, Hedi; Shi, Haifeng; Chen, Keping; Zhang, Zhiyan; Tan, Xiaoli

    2014-10-01

    Data normalization is essential for reliable output of quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) assays, as the unsuitable choice of reference gene(s), whose expression might be influenced by exogenous treatments in plant tissues, could cause misinterpretation of results. To date, no systematic studies on reference genes have been performed in stressed Brassica napus. In this study, we investigated the expression variations of nine candidate reference genes in 40 samples of B. napus leaves subjected to various exogenous treatments. Parallel analyses by geNorm and NormFinder revealed that optimal reference genes differed across the different sets of samples. The best-ranked reference genes were PP2A and TIP41 for salt stress, TIP41 and ACT7 for heavy metal (Cr(6+)) stress, PP2A and UBC21 for drought stress, F-box and SAND for cold stress, F-box and ZNF for salicylic acid stress, TIP41, ACT7, and PP2A for methyl jasmonate stress, TIP41 and ACT7 for abscisic acid stress, and TIP41, UBC21, and PP2A for Sclerotinia sclerotiorum stress. Two newly employed reference genes, TIP41 and PP2A, showed better performances, suggesting their suitability in multiple conditions. To further validate the suitability of the reference genes, the expression patterns of BnWRKY40 and BnMKS1 were studied in parallel. This study is the first systematic analysis of reference gene selection for qRT-PCR normalization in B. napus, an agriculturally important crop, under different stress conditions. The results will contribute toward more accurate and widespread use of qRT-PCR in gene analysis of the genus Brassica.

  6. Solving the maxwell equations by the Chebyshev method : A one-step finite-difference time-domain algorithm

    NARCIS (Netherlands)

    De Raedt, H; Michielsen, K; Kole, JS; Figge, MT

    2003-01-01

    We present a one-step algorithm that solves the Maxwell equations for systems with spatially varying permittivity and permeability by the Chebyshev method. We demonstrate that this algorithm may be orders of magnitude more efficient than current finite-difference time-domain (FDTD) algorithms.

  7. One-Step Multiplex PCR Assay for Differentiating Proposed New Species "Clostridium neonatale" from Closely Related Species.

    Science.gov (United States)

    Ferraris, Laurent; Schönherr, Sophia; Bouvet, Philippe; Dauphin, Brunhilde; Popoff, Michel; Butel, Marie Jose; Aires, Julio

    2015-11-01

    "Clostridium neonatale" sp. nov., previously involved in an outbreak of neonatal necrotizing enterocolitis, was recently proposed as a new species of the Clostridium genus sensu stricto. We developed a one-step multiplex colony PCR for C. neonatale identification and investigated C. neonatale intestinal colonization frequency in healthy preterm neonates. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  8. Direct construction of predictive models for describing growth Salmonella enteritidis in liquid eggs – a one-step approach

    Science.gov (United States)

    The objective of this study was to develop a new approach using a one-step approach to directly construct predictive models for describing the growth of Salmonella Enteritidis (SE) in liquid egg white (LEW) and egg yolk (LEY). A five-strain cocktail of SE, induced to resist rifampicin at 100 mg/L, ...

  9. One-step Synthesis of n-Butanol from Ethanol Condensation over Alumina-supported Metal Catalysts

    Institute of Scientific and Technical Information of China (English)

    Ke Wu YANG; Xuan Zhen JIANG; Wei Chao ZHANG

    2004-01-01

    One-step synthesis of n-butanol from bimolecular condensation of ethanol was firstly achieved over nickel supported gamma alumina catalyst. A mechanism of dehydration path for the growth of carbon chain by eliminating a hydroxy group from one ethanol molecule with a α-H of other ethanol molecule rather than aldol condensation was verified.

  10. A novel approach for one-step forming ε-caprolactam from cyclohexane nitrozation catalyzed by transition metal salt

    Institute of Scientific and Technical Information of China (English)

    Li Qiu Mao; BO Hua Wu; Du Lin Yin; Kui Yi You; Ping Le Liu; He An Luo

    2007-01-01

    The nitrozation reaction of cyclohexane in one-step reaction to form ε-caprolactam has been studied using transition metal salt as catalysts in this work. The results indicated that the catalysts play an especially important role. This method is expected to be a novel way to synthesize other lactam by similar reaction. The possible mechanism was suggested.

  11. Estimating Digital Inter-Symbol Interference Channel Blindly Based on the One-Step Branch Transition Rules in Trellises

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    A novel discrete-time digital inter-symbol interference (ISI) channel blind estimation sub-optimal algorithm is proposed. This algorithm reduces the complexity of the optimal maximum likelihood sequence estimation(MLSE) considerably based on the one-step branch transition rules in trellises, and is suitable for the estimation of the channels with small lengths of ISI.

  12. Versatile, Fast, and Easy One-Step Method for the Synthesis of Hydrophilic Lanthanide-Doped Nanoparticles

    NARCIS (Netherlands)

    Grana Suarez, Laura; Verboom, Willem; Sarkar, Shyam; Mahalingam, V.; Huskens, Jurriaan

    2016-01-01

    The preparation of hydrophilic lanthanide-doped nanoparticles (NPs) following a versatile one-step colloidal method at low temperature and short reaction time with different doped lanthanides, capping ligands, and fluoride sources is presented. The photoluminescence of the particles, via both

  13. Treatment of fish processing wastewater in a one-step or two-step upflow anaerobic sludge blanket (UASB) reactor

    NARCIS (Netherlands)

    Paluenzuela-Rollon, A.; Zeeman, G.; Lubberding, H.J.; Lettinga, G.; Alaerts, G.J.

    2002-01-01

    The performance of one-step UASB reactors treating fish processing wastewater of different lipid levels was determined using artificially generated influent simulating that of the canning of sardines and tuna. The organic loading rates (OLR) and the hydraulic retention times (HRT) were 5-8 g COD.l(-

  14. One-Step Generation of Cluster States Assisted by a Strong Driving Classical Field in Cavity Quantum Electrodynamics

    Institute of Scientific and Technical Information of China (English)

    SHAO Xiao-Qiang; ZHANG Shou

    2008-01-01

    We propose a scheme for one-step generation of cluster states with atoms sent through a thermal cavity with strong classical driving field, based on the resonant atom-cavity interaction so that the operating time is sharply short, which is important in the view of decoherence.

  15. Fabrication of an all-polymer electrochemical sensor by using a one-step hot embossing procedure

    DEFF Research Database (Denmark)

    Kafka, Jan Robert; Larsen, Niels Bent; Skaarup, Steen

    2010-01-01

    We present a fast one-step hot embossing procedure for fabricating an all-polymer electrochemical sen¬sor based on a thin, conductive film of poly(3,4-ethylenedioxythiophene) (PEDOT), a few 100s of nano¬meters in thickness, polymerised on top of a non-conductive TOPAS® (Cyclic Olefin Copolymer...

  16. One-step implementation of maximally entangled states of many three-level atoms in microwave cavity QED

    Science.gov (United States)

    Zou, Xubo; Mathis, W.

    2004-09-01

    We propose an experimental scheme for one-step implementation of maximally entangled states of many three-level atoms in microwave cavity QED. In the scheme, many three-level atoms initially prepared in the same superposition states are simultaneously sent through one superconducting cavity, and maximally entangled states can be generated without requiring the measurement and individual addressing of the atoms.

  17. Treatment of fish processing wastewater in a one-step or two-step upflow anaerobic sludge blanket (UASB) reactor

    NARCIS (Netherlands)

    Paluenzuela-Rollon, A.; Zeeman, G.; Lubberding, H.J.; Lettinga, G.; Alaerts, G.J.

    2002-01-01

    The performance of one-step UASB reactors treating fish processing wastewater of different lipid levels was determined using artificially generated influent simulating that of the canning of sardines and tuna. The organic loading rates (OLR) and the hydraulic retention times (HRT) were 5-8 g

  18. Synthesis of Copper Oxalate Nanorods by a Simple One-step Solid-state Chemical Reaction Method

    Institute of Scientific and Technical Information of China (English)

    CAO Ya-li; JIA Dian-zeng; LIU Lang; XIAO Ding-quan; XIN Xin-quan

    2005-01-01

    Copper oxalate nanorods were successfully prepared by means of a simple one-step solid-state reaction method with the assistance of a suitable surfactant, polyethylene glycol 400. The product with uniform rodlike morphology was characterized by XRD, TEM and SEM. The formational mechanism of the rod-like structure was also preliminary discussed.

  19. Rapid detection and differentiation of dengue virus serotypes by NS1 specific reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay in patients presenting to a tertiary care hospital in Hyderabad, India.

    Science.gov (United States)

    Neeraja, M; Lakshmi, V; Lavanya, Vanjari; Priyanka, E N; Parida, M M; Dash, P K; Sharma, Shashi; Rao, P V Lakshmana; Reddy, Gopal

    2015-01-01

    Early and rapid detection of dengue virus (DENV) infection during the acute phase of illness is crucial for proper patient management and prevention of the spread of the infection. In the present study, the standardization and validation of a one step, four tube reverse transcription loop-mediated isothermal amplification assay (RT-LAMP) for rapid detection and serotyping of the DENV targeting NS1 gene using the Genie® II flourometer was carried out. The performance of the RT-LAMP was compared to RT-PCR, CDC 1-4 Real time PCR and the NS1 antigen ELISA, IgM and IgG anti DENV antibodies. Acute DENV infection was confirmed in 250/300 patients suspected clinically of DENV infection. RT- LAMP and CDC 1-4 Real time PCR assay was positive in 148/250 patients, while 92/250 patients were positive for anti- Dengue IgM and IgG antibodies. The RT-LAMP assay and the CDC real-time RT-PCR assay showed high concordance (k=1.0). The detection rate of acute DENV infection improved to 96% (240/250) when the results of RT-LAMP were combined with NS1 Ag, IgM and IgG ELISA. The RT-LAMP had a detection limit of 100 copies for DEN-1 and DEN-2, 10 copies for DEN-3 and DEN-4 compared to 1000 copies for DEN-1 and DEN-2, 100 copies for DEN-3 and DEN-4 by the conventional RT-PCR. The assay showed 100% specificity. The RT-LAMP assay developed in this study has potential use for early clinical diagnosis, serotyping and surveillance of DENV infection in endemic countries such as India.

  20. Imidazolium ionic liquid induced one-step synthesis of -Fe2O3 nanorods and nanorod assemblies for lithium-ion battery

    Directory of Open Access Journals (Sweden)

    Shuting Xie

    2016-12-01

    Full Text Available α-Fe2O3 nanorods and nanorod assemblies are prepared via a facile one-step method with the assistance of imidazolium-based ionic liquid. The aspect ratio of synthesized nanorods is determined by the alkyl chain length of [Cnmim]+. The inter-molecular π−π interaction and intra-molecular dipole-dipole interaction among imidazole rings of [C4mim]+[PhCOO]− play critical roles in both nucleation and assembly processes of α-Fe2O3 nanorods. The α-Fe2O3 nanorod assemblies show an excellent performance in lithium-ion batteries with a reversible capacity of 1007.3 mA h g−1 at the rate of 500 mA g−1 after 150 cycles.

  1. Detection limits of several commercial reverse transcriptase enzymes: impact on the low- and high-abundance transcript levels assessed by quantitative RT-PCR

    Directory of Open Access Journals (Sweden)

    Delbecchi Louis

    2007-10-01

    Full Text Available Abstract Background In functional genomics, transcript measurement is of fundamental importance. Quantitative reverse transcription polymerase chain reaction (qRT-PCR assays are the most popular technology and depend on the initial molecular step, the reverse transcription (RT. This study provides a complex overview of the influence of elements such as RT systems, amount of background RNA, and transcript abundance on the efficiency of qRT-PCR. Using qRT-PCR, we compared the efficiency of some commonly used RT systems and measured the production of PCR-amplifiable products and the influence of PCR inhibitor contents. Results The qRT-PCR assays were conducted using the TaqMan system, although we also tested the SYBR Green I chemistry, which is not compatible with all the RT systems. When dealing with low-abundance transcripts, the SuperScript II system generated more detectable molecules than the four other systems tested: Sensiscript, Omniscript, SuperScript III and PowerScript (P P Conclusion This study provides a complex overview of the influence of elements such as RT systems, qRTPCR chemistry, amount of background RNA, and transcript abundance on the efficiency of qRT-PCR. Whereas the most significant influencing factor is the presence of inhibitors in the RT systems, total background RNA is also a major influencing component that affects the PCR reaction. Whenever the aim of a study is to obtain a precise gene expression measurement or to profile the global transcriptome (e.g. microarray, the RT step is critical and should be examined with care.

  2. Simultaneous detection and differentiation of three genotypes of Brassica yellows virus by multiplex reverse transcription-polymerase chain reaction.

    Science.gov (United States)

    Zhang, Xiaoyan; Peng, Yanmei; Wang, Ying; Zhang, Zongying; Li, Dawei; Yu, Jialin; Han, Chenggui

    2016-11-22

    Brassica yellows virus (BrYV), proposed to be a new polerovirus species, three distinct genotypes (BrYV-A, BrYV-B and BrYV-C) have been described. This study was to develop a simple, rapid, sensitive, cost-effective method for simultaneous detection and differentiation of three genotypes of BrYV. In this study, a multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was developed for simultaneous detection and differentiation of the three genotypes of BrYV. The three genotypes of BrYV and Tunip yellows virus (TuYV) could be differentiated simultaneously using six optimized specific oligonucleotide primers, including one universal primer for detecting BrYV, three BrYV genotype-specific primers, and a pair of primers for specific detection of TuYV. Primers were designed from conserved regions of each virus and their specificity was confirmed by sequencing PCR products. The mRT-PCR products were 278 bp for BrYV-A, 674 bp for BrYV-B, 505 bp for BrYV-C, and 205 bp for TuYV. Amplification of three target genotypes was optimized by increasing the PCR annealing temperatures to 62 °C. One to three fragments specific for the virus genotypes were simultaneously amplified from infected samples and identified by their specific molecular sizes in agarose gel electrophoresis. No specific products could be amplified from cDNAs of other viruses which could infect crucifer crops. Detection limits of the plasmids for multiplex PCR were 100 fg for BrYV-A and BrYV-B, 10 pg for BrYV-C, and 1 pg for TuYV, respectively. The mRT-PCR was applied successfully for detection of three BrYV genotypes from field samples collected in China. The simple, rapid, sensitive, and cost-effective mRT-PCR was developed successfully for detection and differentiation of the three genotypes of BrYV.

  3. Screening suitable reference genes for normalization in reverse transcription quantitative real-time PCR analysis in melon.

    Directory of Open Access Journals (Sweden)

    Qiusheng Kong

    Full Text Available Melon (Cucumis melo. L is not only an economically important cucurbitaceous crop but also an attractive model for studying many biological characteristics. Screening appropriate reference genes is essential to reverse transcription quantitative real-time PCR (RT-qPCR, which is key to many studies involving gene expression analysis. In this study, 14 candidate reference genes were selected, and the variations in their expression in roots and leaves of plants subjected to biotic stress, abiotic stress, and plant growth regulator treatment were assessed by RT-qPCR. The stability of the expression of the selected genes was determined and ranked using geNorm and NormFinder. geNorm identified the two most stable genes for each set of conditions: CmADP and CmUBIep across all samples, CmUBIep and CmRPL in roots, CmRAN and CmACT in leaves, CmADP and CmRPL under abiotic stress conditions, CmTUA and CmACT under biotic stress conditions, and CmRAN and CmACT under plant growth regulator treatments. NormFinder determined CmRPL to be the best reference gene in roots and under biotic stress conditions and CmADP under the other experimental conditions. CmUBC2 and CmPP2A were not found to be suitable under many experimental conditions. The catalase family genes CmCAT1, CmCAT2, and CmCAT3 were identified in melon genome and used as target genes to validate the reliability of identified reference genes. The catalase family genes showed the most upregulation 3 days after inoculation with Fusarium wilt in roots, after which they were downregulated. Their levels of expression were significantly overestimated when the unsuitable reference gene was used for normalization. These results not only provide guidelines for the selection of reference genes for gene expression analyses in melons but may also provide valuable information for studying the functions of catalase family genes in stress responses.

  4. Improving the tensile strength of carbon nanotube yarn via one-step double [2+1] cycloadditions

    Energy Technology Data Exchange (ETDEWEB)

    Kim, HeeJin [Kyungpook National University, Daegu (Korea, Republic of); Lee, Jaegeun; Park, Byungrak; Sa, Jeong Hoon; Jung, Alum; Kim, Teawon; Park, Junbeom; Hwang, Woonbong; Lee, Kun Hong [Pohang University of Science and Technology, Pohang (Korea, Republic of)

    2016-01-15

    The tensile strength of a CNT yarn was improved through simple one-step double [2+1] cycloaddition reactions that crosslinked the constituent CNTs using a polyethylene glycol (PEG)-diazide crosslinker. The FT-IR spectrum confirmed that the azide groups in the PEG-diazide were converted into aziridine rings, indicating that the cycloaddition reaction was successful. The generation of crosslinked CNTs was also supported by the observation of N1s peak in the XPS spectrum and the increased thermal stability of the material, as observed by TGA. The tensile strength of the CNT yarn was increased from 0.2GPa to 1.4GPa after the crosslinking reaction when twisted at 4000 twists/ meter. The appropriate selection of the crosslinker may further optimize the CNT yarn crosslinking reaction. The simplicity of this one-step crosslinking reaction provides an economical approach to the mass production of high-strength CNT yarns.

  5. Rapid One-Step Selection Method for Generating Nucleic Acid Aptamers: Development of a DNA Aptamer against alpha-Bungarotoxin

    DEFF Research Database (Denmark)

    Lauridsen, Lasse Holm; Shamaileh, Hadi A.; Edwards, Stacey L.

    2012-01-01

    Background: Nucleic acids based therapeutic approaches have gained significant interest in recent years towards the development of therapeutics against many diseases. Recently, research on aptamers led to the marketing of Macugen (R), an inhibitor of vascular endothelial growth factor (VEGF...... by PCR enrichment of the selected aptamers. One round of selection successfully identified a DNA aptamer sequence with a binding affinity of 7.58 mu M. Conclusion: We have demonstrated a one-step method for rapid production of nucleic acid aptamers. Although the reported binding affinity is in the low...... micromolar range, we believe that this could be further improved by using larger targets, increasing the stringency of selection and also by combining a capillary electrophoresis separation prior to the one-step selection. Furthermore, the method presented here is a user-friendly, cheap and an easy way...

  6. Linear Unbiased Optimal Filter for Discrete-Time Systems with One-Step Random Delays and Inconsecutive Packet Dropouts

    Directory of Open Access Journals (Sweden)

    Jian Ding

    2013-11-01

    Full Text Available This paper is concerned with the linear unbiased minimum variance estimation problem for discrete-time stochastic linear control systems with one-step random delay and inconsecutive packet dropout. A new model is developed to describe the phenomena of the one-step delay and inconsecutive packet dropout by employing a Bernoulli distributed stochastic variable. Based on the model, a recursive linear unbiased optimal filter in the linear minimum variance sense is designed by the method of completing the square. The solution to the linear filter is given by three equations including a Riccati equation, a Lyapunov equation and a simple difference equation. A sufficient condition for the existence of the steady-state filter is given. A simulation shows the effectiveness of the proposed algorithm.    

  7. Sheets of branched poly(lactic acid) obtained by one step reactive extrusion calendering process: Melt rheology analysis

    OpenAIRE

    Cailloux, J.; Santana, O.O.; E. Franco-Urquiza; Bou, J. J.; F. Carrasco; J. Gamez-Perez; M. L. Maspoch

    2013-01-01

    One-step reactive extrusion-calendering process (REX-Calendering) was used in order to obtain sheets of 1mm from two PD,L-LA extrusion grades modified with a styrene-acrylic multifunctional oligomeric agent. In a preliminary internal mixer study, torque versus time was monitored in order to determine chain extender ratios and reaction time. Once all parameters were optimized, reactive extrusion experiments were performed. Independently of the processing method employed, under the same pro...

  8. Quantum dot-enhanced detection of dual short RNA sequences via one-step template-dependent surface hybridization

    Energy Technology Data Exchange (ETDEWEB)

    Song Wenqing; Qiu Xue; Lau Choiwan [School of Pharmacy, Key Laboratory of Smart Drug Delivery, Ministry of Education and PLA, Fudan University, 826 Zhangheng Road, Shanghai 201203 (China); Lu Jianzhong, E-mail: jzlu@shmu.edu.cn [School of Pharmacy, Key Laboratory of Smart Drug Delivery, Ministry of Education and PLA, Fudan University, 826 Zhangheng Road, Shanghai 201203 (China)

    2012-07-20

    Graphical abstract: A novel multiplexed method for short RNA detection is reported that employed a design strategy in which quantum dots functionalized reporter DNA were used to capture a short single-stranded RNA sequence from a target solution and then to specifically adsorb onto a common capture probe-modified 96-well plate via a one-step template-dependent, surface hybridization for simultaneous fluorescence detection. Highlights: Black-Right-Pointing-Pointer A ligase-free sensor was demonstrated for the specific detection of dual short RNA. Black-Right-Pointing-Pointer The method was sensitive and simultaneous. Black-Right-Pointing-Pointer Quantum dots-modified reporter probes could increase the melting temperature. Black-Right-Pointing-Pointer Quantum dots functionalized reporter DNA hybridized with capture DNA and target RNA. Black-Right-Pointing-Pointer Target RNA was captured via a one-step template-dependent hybridization. - Abstract: A novel multiplexed method for short RNA detection is reported that employs a design strategy in which capture and reporter probes anneal to each other in the presence of a short RNA target via the formation of a stable three-component complex. Quantum dots (QDs) functionalized with reporter DNA are thus specifically bound onto a capture probe-modified 96-well plate by one-step hybridization for simple RNA detection. In comparison with conventional organic dye-modified reporter probes, the use of reporter DNA-modified QD conjugates increase the melting temperature and lead to the detection of short RNA without the need for a ligation reaction. Moreover, QD properties allow multiple short RNA sequences to be simultaneously determined via rapid and simple one-step hybridization, as exemplified herein. The present results clearly demonstrate that this new strategy can be used to detect dual-short RNA sequence at concentrations of 10 pM in 100 {mu}L.

  9. Multiple Coatings can Improve the Bond Durability of One-step Self-etching Adhesive to Primary Dentin

    Institute of Scientific and Technical Information of China (English)

    Lin Ma; Jian-feng Zhou; Quan Jing; Ji-zhi Zhao; Kuo Wan

    2012-01-01

    Objective To investigate whether multiple coatings can improve the bond durability of one-step self-etching adhesive to primary dentin.Methods Twelve caries-free human primary molars were randomly divided into 2 groups.In group 1,each tooth was hemisected into 2 halves.One half was assigned to the control subgroup 1,which was bonded with a commercially available one-step self-etching adhesive according to the manufacturer's instructions; the other half was assigned to experimental subgroup 1,in which the adhesive was applied three times before light curing.In group 2,one split half tooth was bonded with a commercially available one-step self-etching adhesive according to the manufacturer's instructions; for the other half,three layers of adhesive were applied with each successive layer of light curing.Specimens were stored in 0.9% NaCl containing 0.02% sodium azide at 37℃ for 18 months and then were subjected to microtensile bond strength test and the fracture mode analysis.Results When the adhesive was applied three times before light curing,the bond strength of the experimental subgroup 1 was significandy higher than that of the control subgroup 1 (47.46±13.91 vs.38.12±11.21 MPa,P<0.05).When using the technique of applying multiple layers of adhesive with each successive layer of light curing,no difference was observed in bond strength between the control subgroup and the experimental subgroup (39.40±8.87 vs.40.87±9.33 MPa,P>0.05).Conclusion Multiple coatings can improve the bond durability of one-step self-etching adhesive to primary dentin when using the technique of light-curing after applying 3 layers of adhesive.

  10. Rapid one-step selection method for generating nucleic acid aptamers: development of a DNA aptamer against α-bungarotoxin.

    Directory of Open Access Journals (Sweden)

    Lasse H Lauridsen

    Full Text Available BACKGROUND: Nucleic acids based therapeutic approaches have gained significant interest in recent years towards the development of therapeutics against many diseases. Recently, research on aptamers led to the marketing of Macugen®, an inhibitor of vascular endothelial growth factor (VEGF for the treatment of age related macular degeneration (AMD. Aptamer technology may prove useful as a therapeutic alternative against an array of human maladies. Considering the increased interest in aptamer technology globally that rival antibody mediated therapeutic approaches, a simplified selection, possibly in one-step, technique is required for developing aptamers in limited time period. PRINCIPAL FINDINGS: Herein, we present a simple one-step selection of DNA aptamers against α-bungarotoxin. A toxin immobilized glass coverslip was subjected to nucleic acid pool binding and extensive washing followed by PCR enrichment of the selected aptamers. One round of selection successfully identified a DNA aptamer sequence with a binding affinity of 7.58 µM. CONCLUSION: We have demonstrated a one-step method for rapid production of nucleic acid aptamers. Although the reported binding affinity is in the low micromolar range, we believe that this could be further improved by using larger targets, increasing the stringency of selection and also by combining a capillary electrophoresis separation prior to the one-step selection. Furthermore, the method presented here is a user-friendly, cheap and an easy way of deriving an aptamer unlike the time consuming conventional SELEX-based approach. The most important application of this method is that chemically-modified nucleic acid libraries can also be used for aptamer selection as it requires only one enzymatic step. This method could equally be suitable for developing RNA aptamers.

  11. Rapid one-step selection method for generating nucleic acid aptamers: development of a DNA aptamer against α-bungarotoxin.

    Science.gov (United States)

    Lauridsen, Lasse H; Shamaileh, Hadi A; Edwards, Stacey L; Taran, Elena; Veedu, Rakesh N

    2012-01-01

    Nucleic acids based therapeutic approaches have gained significant interest in recent years towards the development of therapeutics against many diseases. Recently, research on aptamers led to the marketing of Macugen®, an inhibitor of vascular endothelial growth factor (VEGF) for the treatment of age related macular degeneration (AMD). Aptamer technology may prove useful as a therapeutic alternative against an array of human maladies. Considering the increased interest in aptamer technology globally that rival antibody mediated therapeutic approaches, a simplified selection, possibly in one-step, technique is required for developing aptamers in limited time period. Herein, we present a simple one-step selection of DNA aptamers against α-bungarotoxin. A toxin immobilized glass coverslip was subjected to nucleic acid pool binding and extensive washing followed by PCR enrichment of the selected aptamers. One round of selection successfully identified a DNA aptamer sequence with a binding affinity of 7.58 µM. We have demonstrated a one-step method for rapid production of nucleic acid aptamers. Although the reported binding affinity is in the low micromolar range, we believe that this could be further improved by using larger targets, increasing the stringency of selection and also by combining a capillary electrophoresis separation prior to the one-step selection. Furthermore, the method presented here is a user-friendly, cheap and an easy way of deriving an aptamer unlike the time consuming conventional SELEX-based approach. The most important application of this method is that chemically-modified nucleic acid libraries can also be used for aptamer selection as it requires only one enzymatic step. This method could equally be suitable for developing RNA aptamers.

  12. One-Step Targeted Minimum Loss-based Estimation Based on Universal Least Favorable One-Dimensional Submodels.

    Science.gov (United States)

    van der Laan, Mark; Gruber, Susan

    2016-05-01

    Consider a study in which one observes n independent and identically distributed random variables whose probability distribution is known to be an element of a particular statistical model, and one is concerned with estimation of a particular real valued pathwise differentiable target parameter of this data probability distribution. The targeted maximum likelihood estimator (TMLE) is an asymptotically efficient substitution estimator obtained by constructing a so called least favorable parametric submodel through an initial estimator with score, at zero fluctuation of the initial estimator, that spans the efficient influence curve, and iteratively maximizing the corresponding parametric likelihood till no more updates occur, at which point the updated initial estimator solves the so called efficient influence curve equation. In this article we construct a one-dimensional universal least favorable submodel for which the TMLE only takes one step, and thereby requires minimal extra data fitting to achieve its goal of solving the efficient influence curve equation. We generalize these to universal least favorable submodels through the relevant part of the data distribution as required for targeted minimum loss-based estimation. Finally, remarkably, given a multidimensional target parameter, we develop a universal canonical one-dimensional submodel such that the one-step TMLE, only maximizing the log-likelihood over a univariate parameter, solves the multivariate efficient influence curve equation. This allows us to construct a one-step TMLE based on a one-dimensional parametric submodel through the initial estimator, that solves any multivariate desired set of estimating equations.

  13. Identification and evaluation of suitable reference genes for gene expression studies in the whitefly Bemisia tabaci (Asia I) by reverse transcription quantitative realtime PCR.

    Science.gov (United States)

    Collins, Carl; Patel, Mitulkumar V; Colvin, John; Bailey, David; Seal, Susan

    2014-05-02

    This study presents a reliable method for performing reverse transcription quantitative realtime PCR (RT-qPCR) to measure gene expression in the whitefly Bemisia tabaci (Asia I) (Gennadius) (Hemiptera: Aleyrodidae), utilising suitable reference genes for data normalisation. We identified orthologs of commonly used reference genes (actin (ACT), cyclophilin 1 (CYP1), elongation factor 1α (EF1A), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), ribosomal protein L13a (RPL13A), and α-tubulin (TUB1A)), measured the levels of their transcripts by RT-qPCR during development and in response to thermal stress, and evaluated their suitability as endogenous controls using geNorm, BestKeeper, and NormFinder programs. Overall, TUB1A, RPL13A, and CYP1 were the most stable reference genes during B. tabaci development, and TUB1A, GAPDH, and RPL13A were the most stable reference genes in the context of thermal stress. An analysis of the effects of reference gene choice on the transcript profile of a developmentally-regulated gene encoding vitellogenin demonstrated the importance of selecting the correct endogenous controls for RT-qPCR studies. We propose the use of TUB1A, RPL13A, and CYP1 as endogenous controls for transcript profiling studies of B. tabaci development, whereas the combination of TUB1A, GAPDH, and RPL13A should be employed for studies into thermal stress. The data pre- sented here will assist future transcript profiling studies in whiteflies.

  14. Identification and Evaluation of Suitable Reference Genes for Gene Expression Studies in the Whitefly Bemisia tabaci (Asia I) by Reverse Transcription Quantitative Real-Time PCR

    Science.gov (United States)

    Collins, Carl; Patel, Mitulkumar V.; Colvin, John; Bailey, David; Seal, Susan

    2014-01-01

    This study presents a reliable method for performing reverse transcription quantitative real-time PCR (RT-qPCR) to measure gene expression in the whitefly Bemisia tabaci (Asia I) (Gennadius) (Hemiptera: Aleyrodidae), utilising suitable reference genes for data normalisation. We identified orthologs of commonly used reference genes (actin (ACT), cyclophilin 1 (CYP1), elongation factor 1α (EF1A), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), ribosomal protein L13a (RPL13A), and α-tubulin (TUB1A)), measured the levels of their transcripts by RT-qPCR during development and in response to thermal stress, and evaluated their suitability as endogenous controls using geNorm, BestKeeper, and NormFinder programs. Overall, TUB1A, RPL13A, and CYP1 were the most stable reference genes during B. tabaci development, and TUB1A, GAPDH, and RPL13A were the most stable reference genes in the context of thermal stress. An analysis of the effects of reference gene choice on the transcript profile of a developmentally-regulated gene encoding vitellogenin demonstrated the importance of selecting the correct endogenous controls for RT-qPCR studies. We propose the use of TUB1A, RPL13A, and CYP1 as endogenous controls for transcript profiling studies of B. tabaci development, whereas the combination of TUB1A, GAPDH, and RPL13A should be employed for studies into thermal stress. The data presented here will assist future transcript profiling studies in whiteflies. PMID:25373210

  15. One Step Further

    DEFF Research Database (Denmark)

    Hansen, Finn Thorbjørn

    2012-01-01

    The phenomenological attitude is essential for practising phenomenology. Many refer to wonder and wonderment as basic attitudes and ways of being present with and listening to phenomena. In this article a critical view is placed on the typically psychologically-loaded language and tonality that i...

  16. Lipopolysaccharide-induced inhibition of transcription of tlr4 in vitro is reversed by dexamethasone and correlates with presence of conserved NFκB binding sites

    Energy Technology Data Exchange (ETDEWEB)

    Bonin, Camila P., E-mail: mila_bonin@yahoo.com.br [Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo 05508-900 (Brazil); Baccarin, Raquel Y.A., E-mail: baccarin@usp.br [Department of Clinics, School of Veterinary Medicine, University of São Paulo, São Paulo 05508-900 (Brazil); Nostell, Katarina, E-mail: katarina.nostell@slu.se [Department of Clinical Sciences, Swedish University of Agricultural Sciences, Box 7054, 750 07 Uppsala (Sweden); Nahum, Laila A., E-mail: laila@nahum.com.br [Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, Belo Horizonte 30190-002 (Brazil); Faculdade Infórium de Tecnologia, Belo Horizonte 30130-180 (Brazil); Fossum, Caroline, E-mail: caroline.fossum@bvf.slu.se [Department of Biomedicine and Veterinary Public Health, Section for Immunology, Swedish University of Agricultural Sciences, BMC, Box 588, SE 751 23 Uppsala (Sweden); Camargo, Maristela M. de, E-mail: mmcamar@usp.br [Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo 05508-900 (Brazil)

    2013-03-08

    Highlights: ► Chimpanzees, horses and humans have regions of similarity on TLR4 and MD2 promoters. ► Rodents have few regions of similarity on TLR4 promoter when compared to primates. ► Conserved NFkB binding sites were found in the promoters of TLR4 and MD2. ► LPS-induced inhibition of TLR4 transcription is reversed by dexamethasone. ► LPS-induced transcription of MD2 is inhibited by dexamethasone. -- Abstract: Engagement of Toll-like receptor 4 (TLR4) by lipopolysaccharide (LPS) is a master trigger of the deleterious effects of septic shock. Horses and humans are considered the most sensitive species to septic shock, but the mechanisms explaining these phenomena remain elusive. Analysis of tlr4 promoters revealed high similarity among LPS-sensitive species (human, chimpanzee, and horse) and low similarity with LPS-resistant species (mouse and rat). Four conserved nuclear factor kappa B (NFκB) binding sites were found in the tlr4 promoter and two in the md2 promoter sequences that are likely to be targets for dexamethasone regulation. In vitro treatment of equine peripheral blood mononuclear cells (eqPBMC) with LPS decreased transcripts of tlr4 and increased transcription of md2 (myeloid differentiation factor 2) and cd14 (cluster of differentiation 14). Treatment with dexamethasone rescued transcription of tlr4 after LPS inhibition. LPS-induced transcription of md2 was inhibited in the presence of dexamethasone. Dexamethasone alone did not affect transcription of tlr4 and md2.

  17. Selenolthiol and dithiol C-terminal tetrapeptide motifs for one-step purification and labeling of recombinant proteins produced in E. coli.

    Science.gov (United States)

    Cheng, Qing; Johansson, Linda; Thorell, Jan-Olov; Fredriksson, Anna; Samén, Erik; Stone-Elander, Sharon; Arnér, Elias S J

    2006-12-01

    We have previously shown that a redox-active selenocysteine-containing tetrapeptide-Sel-tag (Gly-Cys-Sec-Gly)-can be used as a C-terminal fusion motif for recombinant proteins produced in Escherichia coli. This Sel-tag allows selenolate-targeted one-step purification, as well as fluorescent labeling or radiolabeling either with gamma emitters (75Se) or with positron-emitting radionuclides (11C). Here we have analyzed four different redox-active C-terminal motifs, carrying either dithiol (Gly-Cys-Cys-Gly or Ser-Cys-Cys-Ser) or selenolthiol (Gly-Cys-Sec-Gly or Ser-Cys-Sec-Ser) motifs. Utilizing these different functional motifs with the same recombinant protein (Fel d 1), we were able to assess their relative reactivities and potential usefulness for biotechnological applications. We found that all four redox-active tags could be utilized for efficient one-step purification to provide pure protein from a crude bacterial lysate through reversible binding to phenylarsine oxide sepharose, with yields and purities comparable to those obtained for a His-tagged protein purified by the more common approach with use of a Ni2+ column. For labeling with electrophilic fluorescent or radioactive compounds, however, the selenolthiol motifs were considerably more efficient than their dithiol counterparts. The results thus show that both the selenolthiol- and the dithiol-containing tags can serve as efficient alternatives to His-tags for protein purification, while the selenolthiol motifs offer additional and unique potential for Sec-targeted labeling. It should therefore be possible to utilize these multifunctional tetrapeptide motifs to develop a wide range of novel biotechnological applications based on Sec targeting with electrophilic compounds.

  18. One-step interfacial thiol-ene photopolymerization for metal nanoparticle-decorated microcapsules (MNP@MCs).

    Science.gov (United States)

    Liu, Dandan; Jiang, Xuesong; Yin, Jie

    2014-06-24

    We herein reported a one-step strategy to prepare the noble metal nanoparticle-decorated microcapsules (MNP@MCs) through the interfacial thiol-ene photopolymerization. In the presence of amphiphlic polyhedral oligomeric silsesquioxane (POSS) containing thiol groups (PTPS) as a reactive surfactant and trimethylolpropane triacrylate (TMPTA) as a cross-linker, the oil phase of toluene dissolved with a photoinitiator was emulsified into a water phase containing a metal precursor to form an oil-in-water (O/W) emulsion. Upon irradiation of ultraviolet (UV) light, the thiol-ene photoploymerization and photoreduction at the interface of toluene/water lead to the formation of the cross-linked wall and metal nanoparticles, respectively. A series of gold, silver, and platinum nanoparticle-decorated microcapsules (AuNP@MC, AgNP@MC, and PtNP@MC) were prepared through this one-step interfacial thiol-ene photopolymerization and were characterized carefully by scanning electron microscopy (SEM), transmission electron microscopy (TEM), and atomic force microscopy (AFM). The results revealed that the obtained MNP@MCs were 2.2-2.7 μm in diameter with a wall of 40-70 nm in thickness, which was covered with the metal nanoparticles. The size and amount of metal nanoparticles increased with the increasing concentration of the metal precursor in water. Furthermore, the catalyst performance of AuNP@MC was studied by reduction of aromatic nitro compounds and exhibited the enhanced catalytic activity and good stability in the reduction of hydrophobic nitrophenol. It is believed that this robust, convenient, simple strategy based on the one-step interfacial thiol-ene photopolymerization will provide an important alternative to fabricate the functional metal nanoparticle-modified microcapsules.

  19. Evaluation of transcription levels of inlA, inlB, hly, bsh and prfA genes in Listeria monocytogenes strains using quantitative reverse-transcription PCR and ability of invasion into human CaCo-2 cells.

    Science.gov (United States)

    Tamburro, Manuela; Sammarco, Michela Lucia; Ammendolia, Maria Grazia; Fanelli, Incoronata; Minelli, Fabio; Ripabelli, Giancarlo

    2015-03-01

    Listeria monocytogenes virulence depends on the activity of well-characterized virulence factors. In this study, transcription levels of inlA, inlB, hly, bsh and prfA genes in L. monocytogenes strains, and the ability of invasion into CaCo-2 cells were investigated. Serotyping, multiplex-PCR for serovar identification and restriction fragment analysis of inlA were performed. Transcription levels and invasiveness were evaluated by quantitative reverse-transcription PCR and by in vitro assays, respectively. The isolates were of serovars 1/2a, 4b, 1/2c, 1/2b and 3a. Full-length inlA profiles were found for nine of ten clinical isolates, while five of seven cultures from foods showed truncated profile. The analysis of transcription levels of virulence factors encoding genes demonstrated a substantial inter-strain heterogeneity, with clinical strains showing higher levels for almost all genes than isolates from food. A correlation between transcription levels of inlA and inlB, as well as between bsh and prfA, was observed. Significant differences between clinical strains and food isolates in the invasion of CaCo-2 cells were found. Analysis of gene transcription and invasiveness of human cells suggests different virulence phenotypes among L. monocytogenes populations, and this characterization could be a useful tool for risk assessment purposes and for the development of public health strategies. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  20. Sheets of branched poly(lactic acid) obtained by one step reactive extrusion calendering process: Physical Aging and Fracture Behaviour

    OpenAIRE

    Cailloux, Jonathan; Santana Pérez, Orlando Onofre; FRANCO URQUIZA, EDGAR ADRIAN; Bou Serra, Jordi; Carrasco Alonso, Félix Ángel; Maspoch Rulduà, Mª Lluïsa

    2014-01-01

    The architectural modifications of a linear poly(D,L-Lactide) acid (PD,L-LA) commercial grade were induced by a one-step reactive extrusion-calendering process using a styrene-glycidyl acrylate copolymer as reactive agent. The melt degradation was counteracted by chain extension and branching reactions, leading to a stabilization of the melt properties and an increase in the molecular weight. For such modified samples [poly(lactic acid) (PLA)-reactive extrusion (REX)], the rate of physical ag...

  1. Rapid Synthesis of Lead Oxide Nanorods by One-step Solid-state Chemical Reaction at Room Temperature

    Institute of Scientific and Technical Information of China (English)

    CAO, Ya-Li(曹亚丽); JIA, Dian-Zeng(贾殿赠); LIU, Lang(刘浪); LUO, Jian-Min(骆建敏)

    2004-01-01

    A simple and facile method was reported to synthesize lead oxide nanorods. Nanorods of lead oxide were obtained directly from grinding solid metal salt and sodium hydroxide in agate mortar with the assistance of a suitable nonionic surfactant in only one step, which is different from the result of hydroxide in solution. The product has been characterized by XRD, TEM and SEM. The formation mechanism of rod-like morphology is discussed and the surfactant plays an important soft-template role in modifying the interface of solid-state reaction and according process of rod-formation.

  2. Large-scale fabrication of In2S3 porous films via one-step hydrothermal process.

    Science.gov (United States)

    Chen, Fei; Deng, Dan; Lei, Yinlin

    2013-10-01

    Large-scale indium sulfide (In2S3) porous films were fabricated via a facile one-step and non-template hydrothermal process using L-cysteine as a capping agent. The impact of reaction conditions such as reaction time, temperatures, and capping agents on the synthesis of the In2S3 porous films were studied. The morphology, structure, and phase composition of In2S3 porous films were characterized by X-ray diffraction (XRD), field-emission scanning electron microscopy (FESEM), and transmission electron microscopy (TEM). The formation process and the optical property of the In2S3 porous films were also evaluated.

  3. One-Step Realization of SWAP Gate with Superconducting Quantum-Interference Devices and Atoms in Cavity QED

    Institute of Scientific and Technical Information of China (English)

    ZHAN Zhi-Ming

    2008-01-01

    We put forward a simple scheme for one-step realization of a two-qubit SWAP gate with SQUIDs (super-conducting quantum-interference devices) in cavity QED via Raman transition. In this scheme, the cavity field is only virtually excited and thus the cavity decay is suppressed. The SWAP gate is realized by using only two lower flux states of the SQUID system and the excited state would not be excited. Therefore, the effect of decoherence caused from the levels of the SQUID system is possibly minimized. The scheme can also be used to implement the SWAP gate with atoms.

  4. One-Step UV-Induced Synthesis of Polypyrrole/Ag Nanocomposites at the Water/Ionic Liquid Interface

    Directory of Open Access Journals (Sweden)

    Yang Xiaoming

    2009-01-01

    Full Text Available Abstract Polpyrrole (PPy/Ag nanocomposites were successfully synthesized at the interface of water and ionic liquid by one-step UV-induced polymerization. Highly dispersed PPy/Ag nanoparticles were obtained by controlling the experimental conditions. The results of Fourier-transform infrared spectroscopy, X-ray diffraction, transmission electron microscopy and X-ray photoelectron spectroscopy revealed that the UV-induced interface polymerization leaded to the formation of PPy incorporating silver nanoparticles. It was also found that the electrical conductivity of PPy/Ag nanocomposite was about 100 times higher than that of pure PPy.

  5. The amphiphilic hydrophobin Vmh2 plays a key role in one step synthesis of hybrid protein-gold nanoparticles.

    Science.gov (United States)

    Politi, Jane; De Stefano, Luca; Longobardi, Sara; Giardina, Paola; Rea, Ilaria; Methivier, Christophe; Pradier, Claire-Marie; Casale, Sandra; Spadavecchia, Jolanda

    2015-12-01

    We report a simple and original method to synthesize gold nanoparticles in which a fungal protein, the hydrophobin Vmh2 from Pleurotus ostreatus and dicarboxylic acid-terminated polyethylene-glycol (PEG) has been used as additional components in a one step process, leading to hybrid protein-metal nanoparticles (NPs). The nanoparticles have been characterized by ultra-violet/visible, infrared and X-ray photoelectron spectroscopies, dynamic light scattering and also by electron microscopy imaging. The results of these analytical techniques highlight nanometric sized, stable, hybrid complexes of about 12 nm, with outer surface rich in functional chemical groups. Interaction with protein and antibodies has also been exploited.

  6. One-step synthesis of collagen hybrid gold nanoparticles and formation on Egyptian-like gold-plated archaeological ivory.

    Science.gov (United States)

    Spadavecchia, Jolanda; Apchain, Emilande; Albéric, Marie; Fontan, Elisabeth; Reiche, Ina

    2014-08-04

    A one-step method is reported to synthesize hybrid gold nanoparticles (AuNPs) by reduction of HAuCl4 in acetic solution in the presence of collagen (Col), dicarboxylic acid-terminated polyethylene glycol (PEG), and cetyltetrammonium bromide (CTAB) mixed with hydoxyapatite (HAP) as surfactants. Such formation process of AuNPs was shown to be responsible for purple stains naturally formed on Egyptianizing archaeological gilded ivories from 8th BC Syria. The understanding of this formation mechanism, which most likely involves a step with hybrid AuNPs, allows the establishing of an authenticity marker of ancient gold-plated ivories.

  7. Changing Safety Culture, One Step at a Time: The Value of the DOE-VPP Program at PNNL

    Energy Technology Data Exchange (ETDEWEB)

    Wright, Patrick A.; Isern, Nancy G.

    2005-02-01

    The primary value of the Pacific Northwest National Laboratory (PNNL) Voluntary Protection Program (VPP) is the ongoing partnership between management and staff committed to change Laboratory safety culture one step at a time. VPP enables PNNL's safety and health program to transcend a top-down, by-the-book approach to safety, and it also raises grassroots safety consciousness by promoting a commitment to safety and health 24 hours a day, 7 days a week. PNNL VPP is a dynamic, evolving program that fosters innovative approaches to continuous improvement in safety and health performance at the Laboratory.

  8. A One-Step-Ahead Smoothing-Based Joint Ensemble Kalman Filter for State-Parameter Estimation of Hydrological Models

    KAUST Repository

    El Gharamti, Mohamad

    2015-11-26

    The ensemble Kalman filter (EnKF) recursively integrates field data into simulation models to obtain a better characterization of the model’s state and parameters. These are generally estimated following a state-parameters joint augmentation strategy. In this study, we introduce a new smoothing-based joint EnKF scheme, in which we introduce a one-step-ahead smoothing of the state before updating the parameters. Numerical experiments are performed with a two-dimensional synthetic subsurface contaminant transport model. The improved performance of the proposed joint EnKF scheme compared to the standard joint EnKF compensates for the modest increase in the computational cost.

  9. One-step flame synthesis of an active Pt/TiO2 catalyst for SO2 oxidation

    DEFF Research Database (Denmark)

    Johannessen, Tue; Koutsopoulos, Sotiris

    2002-01-01

    size of the platinum particles supported on aggregated nano-particles of TiO2 is approximately 2 nm. The high SO2-oxidation activity of the catalyst proves that platinum is not hidden in the titania matrix. The flame-produced catalyst showed catalytic activity similar to samples prepared by wet......Flame synthesis as a route for production of composite metal oxides has been employed for the one-step synthesis of a supported noble metal catalyst, i.e. a Pt/TiO2 catalyst, by simultaneous combustion of Ti-isopropoxide and platinum acetylacetonate in a quench-cooled flame reactor. The average...

  10. Evidence of a one-step replica symmetry breaking in a three-dimensional Potts glass model.

    Science.gov (United States)

    Takahashi, Takashi; Hukushima, Koji

    2015-02-01

    We study a seven-state Potts glass model in three dimensions with first-, second-, and third-nearest-neighbor interactions with a bimodal distribution of couplings by Monte Carlo simulations. Our results show the existence of a spin-glass transition at a finite temperature T(c), a discontinuous jump of an order parameter at T(c) without latent heat, and a nontrivial structure in the order parameter distribution below T(c). They are compatible with one-step replica symmetry breaking.

  11. A One-Step PCR Method for Detecting the First Base of Splice Donor of Wx Intron 1 in Rice

    Institute of Scientific and Technical Information of China (English)

    MAO Xing-xue; LIU Yan-zhuo; XIAO Xin; CHEN Jian-wei; LUO Wen-yong; LI Xiao-fang

    2004-01-01

    A new method of one-step PGR was devised for detecting the first nucleotide in the splice donor site of Wx intron 1.compared to the regular PCR-Acc I method, the method can produce the same result for detecting +1 nucleotide of Wx intron 1.The reliability of the new method was confirmed with 30 rice varieties. The new technique is more convenient and cheaper than the regular PCR-Acc I method, and could be widely deploded in rice molecular marker assistant selection.

  12. One-step flame synthesis of an active Pt/TiO2 catalyst for SO2 oxidation

    DEFF Research Database (Denmark)

    Johannessen, Tue; Koutsopoulos, Sotiris

    2002-01-01

    Flame synthesis as a route for production of composite metal oxides has been employed for the one-step synthesis of a supported noble metal catalyst, i.e. a Pt/TiO2 catalyst, by simultaneous combustion of Ti-isopropoxide and platinum acetylacetonate in a quench-cooled flame reactor. The average...... size of the platinum particles supported on aggregated nano-particles of TiO2 is approximately 2 nm. The high SO2-oxidation activity of the catalyst proves that platinum is not hidden in the titania matrix. The flame-produced catalyst showed catalytic activity similar to samples prepared by wet...

  13. Development of reverse transcription loop-mediated isothermal amplification assay as a simple detection method of Chrysanthemum stem necrosis virus in chrysanthemum and tomato.

    Science.gov (United States)

    Suzuki, Ryoji; Fukuta, Shiro; Matsumoto, Yuho; Hasegawa, Toru; Kojima, Hiroko; Hotta, Makiko; Miyake, Noriyuki

    2016-10-01

    For a simple and rapid detection of Chrysanthemum stem necrosis virus (CSNV) from chrysanthemum and tomato, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed. A primer set designed to the genome sequences of CSNV worked most efficiently at 63°C and could detect CSNV RNA within 12min by fluorescence monitoring using an isothermal DNA amplification and fluorescence detection device. The result of a specificity test using seven other viruses and one viroid-infectable chrysanthemum or tomato showed that the assay could amplify CSNV specifically, and a sensitivity comparison showed that the RT-LAMP assay was as sensitive as the reverse transcriptase polymerase chain reaction. The RT-LAMP assay using crude RNA, extracted simply, could detect CSNV. Overall, the RT-LAMP assay was found to be a simple, specific, convenient, and time-saving method for CSNV detection.

  14. High-resolution magic angle spinning 1H NMR spectroscopy and reverse transcription-PCR analysis of apoptosis in a rat glioma.

    Science.gov (United States)

    Griffin, Julian L; Blenkiron, Cherie; Valonen, Piia K; Caldas, Carlos; Kauppinen, Risto A

    2006-03-01

    The functional genomic approaches of transcriptomics, proteomics and metabolomics aim to measure the mRNA, protein or metabolite complement of a cell, tissue or organism. In this study we have investigated the compatibility of transcriptional analysis, using Reverse Transcription (RT)-PCR, and metabolite analysis, by high-resolution magic angle spinning (HRMAS) 1H NMR spectroscopy, in BT4C rat glioma following the induction of programmed cell death. The metabolite and transcriptional changes that accompanied apoptosis were examined at 0, 4 and 8 days of ganciclovir/thymidine kinase gene therapy. Despite the high spinning speeds employed during HRMAS 1H NMR spectroscopy of one-half of the tumor samples, RT-PCR analysis of the pro-apoptotic transcripts Bcl-2, BAK-1, caspase-9 and FAS was possible, producing similar results to those detected in the unspun half of the tumors. Furthermore, the expression of FAS was inversely correlated with some of the key metabolic changes across the time period examined including the increases CH=CH and CH=CHCH2 lipid resonances which accompany apoptosis. This study demonstrates how combined transcriptomic and metabolomic studies of tumors can be used to understand the molecular events that accompany well documented metabolic perturbations during cell death processes.

  15. Real-time quantitative reverse transcription-PCR analysis of expression stability of Actinobacillus pleuropneumoniae housekeeping genes during in vitro growth under iron-depleted conditions

    DEFF Research Database (Denmark)

    Nielsen, K. K.; Boye, Mette

    2005-01-01

    The aims of the present investigation were to develop and test a sensitive and reproducible method for the study of gene expression in the porcine lung pathogen Actinobacillus pleuropneumoniae by real-time quantitative reverse transcription (RT)-PCR and to evaluate a number of suitable internal...... up-regulation under iron-restricted conditions compared to bacteria grown in medium with sufficient iron. The observed expression patterns of the genes of interest were consistent with previous observations. This study therefore lends further support to the use of real-time quantitative RT...

  16. Detection and strain differentiation of infectious bronchitis virus in tracheal tissues from experimentally infected chickens by reverse transcription-polymerase chain reaction. Comparison with an immunohistochemical technique

    DEFF Research Database (Denmark)

    Handberg, Kurt; Nielsen, O.L.; Pedersen, M.W.;

    1999-01-01

    Oligonucleotide pairs were constructed for priming the amplification of fragments of nucleocapsid (N) protein and spike glycoprotein (S) genes of avian infectious bronchitis virus (IBV) by reverse transcription-polymerase chain reaction (RT-PCR). One oligonucleotide pair amplified a common segment......3896 and 793B strains of IBV, respectively, Groups of specific pathogen free chickens were experimentally inoculated with the Massachusetts (H120, M41), the D1466 and the 793B strains of IBV, and tracheal tissue preparations were made from each bird for RT-PCR and for immunohistochemistry (IHC) up to 3...

  17. Porcine reproductive and respiratory syndrome virus: Interlaboratory ring trial to evaluate real-time reverse transcription polymerase chain reaction detection methods

    DEFF Research Database (Denmark)

    Wernike, Kerstin; Bonilauri, Paolo; Dauber, Malte

    2012-01-01

    To compare the real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) assays used for the diagnosis of Porcine reproductive and respiratory syndrome virus (PRRSV), a Europe-wide interlaboratory ring trial was conducted. A variety of PRRSV strains including North American...... (NA) and European (EU) genotype isolates were analyzed by the participants. Great differences regarding qualitative diagnostics as well as analytical sensitivity were observed between the individual RT-qPCR systems, especially when investigating strains from the EU genotype. None of the assays...

  18. Multiplex reverse transcription-polymerase chain reaction combined with on-chip electrophoresis as a rapid screening tool for candidate gene sets

    DEFF Research Database (Denmark)

    Wittig, Rainer; Salowsky, Rüdiger; Blaich, Stephanie

    2005-01-01

    Combining multiplex reverse transcription-polymerase chain reaction (mRT-PCR) with microfluidic amplicon analysis, we developed an assay for the rapid and reliable semiquantitative expression screening of 11 candidate genes for drug resistance in human malignant melanoma. The functionality...... of this approach was demonstrated by low interexperimental variations of amplicon quantities after endpoint analysis. When applied to RNA samples derived from drug-sensitive and -resistant melanoma cell lines, mRT-PCR delivered results qualitatively concordant with data obtained from Northern blot and array...

  19. Simple One-Step Synthesis and Superconducting Properties of SmFeAsO1-x Fx

    Institute of Scientific and Technical Information of China (English)

    MA Yan-Wei; GAO Zhao-Shun; WANG Lei; QI Yan-Peng; WANG Dong-Liang; ZHANG Xian-Ping

    2009-01-01

    The recent discovery of superconductivity in REFeAsO (RE,rare-earth metal) has generated enormous interest because these materials are the first non-copper oxide superconductors with critical temperatures Tc exceeding 50 K as well as upper critical fields well above 100 T.However,for these new superconductors,very complicated synthesis routes,such as the complex two-step synthesis or high-pressure sintering,are required.Furthermore,there is the toxicity and volatility of arsenic to consider,sometimes a sealed quartz tube of arsenic exploded during annealing.We present a new method for producing high-temperature SmFeAsO1-xFx superconductors by using a one-step sintering process.Superconducting transition with the onset temperature of 54.6 K and high critical fields Hc2 (0) ≥ 200 T were confirmed in SmFeAsO0.7F0.3.At 5 K and at self field,critical current densities Jc estimated from the magnetization hysteresis using the whole sample size and the average particle size have reached 8.5×103 and 1.2×106 A/cm2,respectively.Moreover,Jc exhibited a very weak dependence on magnetic field. This simple and safe one-step synthesis technique should be effective in other rare earth derivatives of iron-based superconductors.

  20. One-step green synthesis of non-hazardous dicarboxyl cellulose flocculant and its flocculation activity evaluation

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, Hangcheng; Zhang, Yong; Yang, Xiaogang; Liu, Hongyi [The Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, College of Materials and Textiles, Zhejiang Sci-Tech University, Hangzhou 310018 (China); Shao, Lan [Technique Center, Hangzhou Xinhua Group Co., Ltd, Hangzhou 310011 (China); Zhang, Xiumei [The Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, College of Materials and Textiles, Zhejiang Sci-Tech University, Hangzhou 310018 (China); Yao, Juming, E-mail: yaoj@zstu.edu.cn [The Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, College of Materials and Textiles, Zhejiang Sci-Tech University, Hangzhou 310018 (China)

    2015-10-15

    The waste management of used flocculants is a thorny issue in the field of wastewater treatment. To natural cellulose based flocculants, utilization of hazardous cellulose solvent and simplification of synthetic procedure are the two urgent problems needing to be further improved. In this work, a series of natural dicarboxyl cellulose flocculants (DCCs) were one-step synthesized via Schiff-base route. The cellulose solvent (NaOH/Urea solution) was utilized during the synthesis process. The full-biodegradable flocculants avoid causing secondary pollution to environment. The chemical structure and solution property of the DCC products were characterized by FT-IR, {sup 1}H NMR, {sup 13}C NMR, TGA, FESEM, charge density and ζ-potential. Kaolin suspension and effluent from paper mill were selected to evaluate the flocculation activity of the DCCs. Their flocculation performance was compared with that of commercial cationic polyacrylamide and poly aluminium chloride flocculants. The positive results showed that the NaOH/Urea solvent effectively promoted the dialdehyde cellulose (DAC) conversion to DCC in the one-step synthesis reaction. The DCCs with the carboxylate content more than 1 mmol/g exhibited steady flocculation performance to kaolin suspension in the broad pH range from 4 to 10. Its flocculation capacity to the effluent from paper mill also showed excellent.

  1. Properties of a Ni-FUSI gate formed by the EBV method and one-step RTA

    Institute of Scientific and Technical Information of China (English)

    Zhang Youwei; Xu Dawei; Wan Li; Wang Zhongjian; Xia Chao; Cheng Xinhong; Yu Yuehui

    2012-01-01

    Nickel fully silicided (Ni-FUSI) gate material has been fabricated on a HfO2 surface to form a NiFUSI gate/HfO2/Si/A1 (MIS) structure by using an ultra-high vacuum e-beam evaporation (EBV) method followed by a one step rapid thermal annealing (RTA) treatment.X-ray diffraction (XRD) and Raman spectroscopy were used to reveal the microstructures and electrical properties of the MIS structure.Results show that a one step post RTA treatment is enough to promote the full reaction of nickel silicide,compared with multiple RTA treatments.Furthermore,the HfO2 gate dielectric film is sensitive to heat treatment,and multiple RTA treatments can damage the electrical properties of the HfO2 film rather than improve them.By optimization of the sample fabrication technique,the MIS capacitor produces good high-frequency capacitance-voltage curves with a hysteresis of 30 mV,a work function of about 5.44-5.53 eV and leakage current density of only 1.45 × 10-8 A/cm2 at -1 V gate bias.

  2. One-step green synthesis of non-hazardous dicarboxyl cellulose flocculant and its flocculation activity evaluation.

    Science.gov (United States)

    Zhu, Hangcheng; Zhang, Yong; Yang, Xiaogang; Liu, Hongyi; Shao, Lan; Zhang, Xiumei; Yao, Juming

    2015-10-15

    The waste management of used flocculants is a thorny issue in the field of wastewater treatment. To natural cellulose based flocculants, utilization of hazardous cellulose solvent and simplification of synthetic procedure are the two urgent problems needing to be further improved. In this work, a series of natural dicarboxyl cellulose flocculants (DCCs) were one-step synthesized via Schiff-base route. The cellulose solvent (NaOH/Urea solution) was utilized during the synthesis process. The full-biodegradable flocculants avoid causing secondary pollution to environment. The chemical structure and solution property of the DCC products were characterized by FT-IR, (1)H NMR, (13)C NMR, TGA, FESEM, charge density and ζ-potential. Kaolin suspension and effluent from paper mill were selected to evaluate the flocculation activity of the DCCs. Their flocculation performance was compared with that of commercial cationic polyacrylamide and poly aluminium chloride flocculants. The positive results showed that the NaOH/Urea solvent effectively promoted the dialdehyde cellulose (DAC) conversion to DCC in the one-step synthesis reaction. The DCCs with the carboxylate content more than 1 mmol/g exhibited steady flocculation performance to kaolin suspension in the broad pH range from 4 to 10. Its flocculation capacity to the effluent from paper mill also showed excellent.

  3. A one-step bioprocess for production of high-content fructo-oligosaccharides from inulin by yeast.

    Science.gov (United States)

    Wang, Da; Li, Fu-Li; Wang, Shi-An

    2016-10-20

    Commercial fructo-oligosaccharides (FOS) are predominantly produced from sucrose by transfructosylation process that presents a maximum theoretical yield below 0.60gFOSgSucrose(-1). To obtain high-content FOS, costly purification is generally employed. Additionally, high-content FOS can be produced from inulin by using endo-inulinases. However, commercial endo-inulinases have not been extensively used in scale-up production of FOS. In the present study, a one-step bioprocess that integrated endo-inulinase production, FOS fermentation, and non-FOS sugars removal into one reactor was proposed to produce high-content FOS from inulin. The bioprocess was implemented by a recombinant yeast strain JZHΔS-TSC, in which a heterologous endo-inulinase gene was expressed and the inherent invertase gene SUC2 was disrupted. FOS fermentation at 40°C from 200g/L chicory inulin presented the maximun titer, yield, and productivity of 180.2±0.8g/L, 0.9gFOSgInulin(-1), and 7.51±0.03g/L/h, respectively. This study demonstrated that the one-step bioprocess was simple and highly efficient.

  4. Fabrication and characterization of metal-packaged fiber Bragg grating sensor by one-step ultrasonic welding

    Science.gov (United States)

    Zhang, Yumin; Zhu, Lianqing; Luo, Fei; Dong, Mingli; Ding, Xiangdong; He, Wei

    2016-06-01

    A metallic packaging technique of fiber Bragg grating (FBG) sensors is developed for measurement of strain and temperature, and it can be simply achieved via one-step ultrasonic welding. The average strain transfer rate of the metal-packaged sensor is theoretically evaluated by a proposed model aiming at surface-bonded metallic packaging FBG. According to analytical results, the metallic packaging shows higher average strain transfer rate compared with traditional adhesive packaging under the same packaging conditions. Strain tests are performed on an elaborate uniform strength beam for both tensile and compressive strains; strain sensitivities of approximately 1.16 and 1.30 pm/μɛ are obtained for the tensile and compressive situations, respectively. Temperature rising and cooling tests are also executed from 50°C to 200°C, and the sensitivity of temperature is 36.59 pm/°C. All the measurements of strain and temperature exhibit good linearity and stability. These results demonstrate that the metal-packaged sensors can be successfully fabricated by one-step welding technique and provide great promise for long-term and high-precision structural health monitoring.

  5. The energy expenditure of stair climbing one step and two steps at a time: estimations from measures of heart rate.

    Science.gov (United States)

    Halsey, Lewis G; Watkins, David A R; Duggan, Brendan M

    2012-01-01

    Stairway climbing provides a ubiquitous and inconspicuous method of burning calories. While typically two strategies are employed for climbing stairs, climbing one stair step per stride or two steps per stride, research to date has not clarified if there are any differences in energy expenditure between them. Fourteen participants took part in two stair climbing trials whereby measures of heart rate were used to estimate energy expenditure during stairway ascent at speeds chosen by the participants. The relationship between rate of oxygen consumption ([Formula: see text]) and heart rate was calibrated for each participant using an inclined treadmill. The trials involved climbing up and down a 14.05 m high stairway, either ascending one step per stride or ascending two stair steps per stride. Single-step climbing used 8.5±0.1 kcal min(-1), whereas double step climbing used 9.2±0.1 kcal min(-1). These estimations are similar to equivalent measures in all previous studies, which have all directly measured [Formula: see text] The present study findings indicate that (1) treadmill-calibrated heart rate recordings can be used as a valid alternative to respirometry to ascertain rate of energy expenditure during stair climbing; (2) two step climbing invokes a higher rate of energy expenditure; however, one step climbing is energetically more expensive in total over the entirety of a stairway. Therefore to expend the maximum number of calories when climbing a set of stairs the single-step strategy is better.

  6. The energy expenditure of stair climbing one step and two steps at a time: estimations from measures of heart rate.

    Directory of Open Access Journals (Sweden)

    Lewis G Halsey

    Full Text Available Stairway climbing provides a ubiquitous and inconspicuous method of burning calories. While typically two strategies are employed for climbing stairs, climbing one stair step per stride or two steps per stride, research to date has not clarified if there are any differences in energy expenditure between them. Fourteen participants took part in two stair climbing trials whereby measures of heart rate were used to estimate energy expenditure during stairway ascent at speeds chosen by the participants. The relationship between rate of oxygen consumption ([Formula: see text] and heart rate was calibrated for each participant using an inclined treadmill. The trials involved climbing up and down a 14.05 m high stairway, either ascending one step per stride or ascending two stair steps per stride. Single-step climbing used 8.5±0.1 kcal min(-1, whereas double step climbing used 9.2±0.1 kcal min(-1. These estimations are similar to equivalent measures in all previous studies, which have all directly measured [Formula: see text] The present study findings indicate that (1 treadmill-calibrated heart rate recordings can be used as a valid alternative to respirometry to ascertain rate of energy expenditure during stair climbing; (2 two step climbing invokes a higher rate of energy expenditure; however, one step climbing is energetically more expensive in total over the entirety of a stairway. Therefore to expend the maximum number of calories when climbing a set of stairs the single-step strategy is better.

  7. One step phase separation process to fabricate superhydrophobic PVC films and its corrosion prevention for AZ91D magnesium alloy

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Na; Li, Jicheng; Bai, Ningning; Xu, Lan; Li, Qing, E-mail: liqingswu@163.com

    2016-07-15

    Graphical abstract: - Highlights: • Independent superhydrophobic polyvinyl chloride (PVC) film was prepared by phase separation process. • The superhydrophobic PVC film showed excellent stability in acid, alkali and salt corrosive solutions. • This film was prepared on magnesium surface protecting it from corrosion. • This method was simple and universal. - Abstract: A one step, simple fabrication method to prepare independent superhydrophobic polyvinyl chloride (PVC) coating is reported in this paper. The rough surface structure and low surface energy could be simply obtained only by a phase separation process. The independent PVC superhydrophobic film was also applied on AZ91D magnesium alloy. Scanning electron microscopy (SEM), water contact angle measurements, electrochemical test and adhesion tests have been performed to characterize the surface morphology, wettability, anti-corrosion and adhesion strength of independent PVC film and superhydrophobic magnesium alloy respectively. The results indicated that whether it was the PVC film or superhydrophobic magnesium, they show static contact angles higher than 150°, excellent anti-corrosion effect and adhesion strength. We believed that the presented method could provide a straightforward and simple route to fabricate low-cost and anti-corrosion coating on various substrate materials. Moreover, this one step process may find potential application in the field of industry because of its simplicity and universality.

  8. Forensic Identification of Human Blood: comparison of two one-step presumptive tests for blood screening of crime scene samples.

    Directory of Open Access Journals (Sweden)

    Ana Flávia Belchior Andrade

    2014-08-01

    Full Text Available Blood is the most common body fluid found at crime scenes. One-step presumptive tests have been designed as a rapid immunological test for the qualitative detection of human hemoglobin in stool samples (faecal occult blood their usefulness for forensic purposes has been demonstrated before. In this study we compare Hexagon OBTI kit and FOB One-step Bioeasy kit sensitivity in the analysis of diluted blood samples. With Hexagon OBTI, positive test results are achieved in whole blood dilutions up to 1:1.000. Sensitivity decreased with aged samples, if samples were not stored under low temperatures regardless of which presumptive test is used. Whole blood tests must take into consideration that “hook” effect may interfere. Comparing both tests, OBTI Hexagon Kit is more sensible to detect diluted blood, showing a wider detection window in all conditions. This is interesting when analyzing forensic samples as forensic analysts usually do not know about the history of the analyzed sample before its collection.

  9. Effect of Multiple Coatings of One-step Self-etching Adhesive on Microtensile Bond Strength to Primary Dentin

    Institute of Scientific and Technical Information of China (English)

    Lin Ma; Jian-feng Zhou; Jian-guo Tan; Quan Jing; Ji-zhi Zhao; Kuo Wan

    2011-01-01

    Objective To investigate the effect of multiple coatings of the one-step self-etching adhesive on immediate microtensile bond strength to primary dentin.Methods Twelve caries-free human primary molars were randomly divided into 2 groups with 6 teeth each. In group 1,each tooth was hemisected into two halves. One half was assigned to control subgroup 1,which was bonded with a single-step self-etching adhesive according to the manufacturer's instructions; the other half was assigned to experimental subgroup 1 in which the adhesive was applied three times before light curing. In group 2, the teeth were also hemisected into two halves. One half was assigned to control subgroup 2, which was bonded with the single-step self-etching adhesive according to the manufacturer's instructions; the other half was assigned to experimental subgroup 2 in which three layers of adhesive were applied with light curing each successive layer. Microtensile bond strength was immediately tested after specimen preparation.Results When the adhesive was applied three times before light curing, the bond strength of the experimental subgroup 1 (n=33, 57.49±11.61 MPa) was higher than that of the control subgroup 1 (n=31,49.71±11.43 MPa, P0.05).Conclusion multiple coatings of one-step self-etching adhesive can increase the immediate bond strength to primary dentin when using the technique of light-curing after applying three layers of adhesive.

  10. One-step room temperature synthesis of very small γ-Fe{sub 2}O{sub 3} nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Moscoso-Londoño, O. [Laboratorio de Sólidos Amorfos, INTECIN, Facultad de Ingeniería, Universidad de Buenos Aires, C1063ACV, Buenos Aires (Argentina); Carrião, M.S. [Laboratorio de Sólidos Amorfos, INTECIN, Facultad de Ingeniería, Universidad de Buenos Aires, C1063ACV, Buenos Aires (Argentina); Instituto de Física, Campus II, Universidade Federal de Goiás, CEP: 74001-970 Goiânia (GO) (Brazil); Cosio-Castañeda, C. [Laboratorio de Sólidos Amorfos, INTECIN, Facultad de Ingeniería, Universidad de Buenos Aires, C1063ACV, Buenos Aires (Argentina); Departamento de Física y Química Teórica, Facultad de Química, Universidad Nacional Autónoma de México, Ciudad Universitaria, D.F. 04510 (Mexico); Bilovol, V. [Laboratorio de Sólidos Amorfos, INTECIN, Facultad de Ingeniería, Universidad de Buenos Aires, C1063ACV, Buenos Aires (Argentina); Sánchez, R. Martínez [Instituto de Materiales y Reactivos, Universidad de La Habana, CP10400, Ciudad Habana (Cuba); Lede, E.J.; Socolovsky, L.M. [Laboratorio de Sólidos Amorfos, INTECIN, Facultad de Ingeniería, Universidad de Buenos Aires, C1063ACV, Buenos Aires (Argentina); Martínez-García, R., E-mail: rmartinez@fi.uba.ar [Laboratorio de Sólidos Amorfos, INTECIN, Facultad de Ingeniería, Universidad de Buenos Aires, C1063ACV, Buenos Aires (Argentina)

    2013-09-01

    Graphical abstract: - Highlights: • One-step synthesis of 3 nm maghemite nanoparticles is reported. • Maghemite nanoparticles can be synthesized from a ferric solution. • γ-Fe{sub 2}O{sub 3} NPs can be obtained if the precursor has Fe(III) in tetrahedral interstices. • HR-TEM, Mössbauer, XAFS and magnetometry analysis proved the maghemite existence - Abstract: Very small maghemite nanoparticles (∼3 nm) are obtained through a one-step synthesis at room temperature. The fast neutralization reaction of a ferric solution in a basic medium produces an intermediate phase, presumably two-line ferrihydrite, which in oxidizing conditions is transformed to maghemite nanoparticles. The synthesis of maghemite, as final product of the reaction, was characterized by High-Resolution Transmission Electron Microscopy (HR-TEM), X-ray Absorption Fine Structure (XAFS), Mössbauer spectroscopy, and magnetometry. The XAFS technique allowed the analysis of the crystallographic variations into maghemite nanoparticles as a result of modification in its surface/volume ratio. Mössbauer spectroscopy at low temperature (4.2 K) confirms the presence of Fe(III) in tetrahedral and octahedral interstices, in the stoichiometry corresponding to maghemite. The specific magnetization, M vs H (3 K and 300 K, up to 7 T) and temperature dependence of the magnetization (50 Oe by ZFC mode, 2 K ≤ T ≤ 300 K) indicate that maghemite nanoparticles of 3 nm are in superparamagnetic state with a blocking temperature close to 36 K.

  11. Laser ablation-based one-step generation and bio-functionalization of gold nanoparticles conjugated with aptamers

    Directory of Open Access Journals (Sweden)

    Walter Johanna G

    2010-08-01

    Full Text Available Abstract Background Bio-conjugated nanoparticles are important analytical tools with emerging biological and medical applications. In this context, in situ conjugation of nanoparticles with biomolecules via laser ablation in an aqueous media is a highly promising one-step method for the production of functional nanoparticles resulting in highly efficient conjugation. Increased yields are required, particularly considering the conjugation of cost-intensive biomolecules like RNA aptamers. Results Using a DNA aptamer directed against streptavidin, in situ conjugation results in nanoparticles with diameters of approximately 9 nm exhibiting a high aptamer surface density (98 aptamers per nanoparticle and a maximal conjugation efficiency of 40.3%. We have demonstrated the functionality of the aptamer-conjugated nanoparticles using three independent analytical methods, including an agglomeration-based colorimetric assay, and solid-phase assays proving high aptamer activity. To demonstrate the general applicability of the in situ conjugation of gold nanoparticles with aptamers, we have transferred the method to an RNA aptamer directed against prostate-specific membrane antigen (PSMA. Successful detection of PSMA in human prostate cancer tissue was achieved utilizing tissue microarrays. Conclusions In comparison to the conventional generation of bio-conjugated gold nanoparticles using chemical synthesis and subsequent bio-functionalization, the laser-ablation-based in situ conjugation is a rapid, one-step production method. Due to high conjugation efficiency and productivity, in situ conjugation can be easily used for high throughput generation of gold nanoparticles conjugated with valuable biomolecules like aptamers.

  12. Effects of cooling rate on vermicular graphite percentage in a brake drum produced by one-step cored wire injection

    Directory of Open Access Journals (Sweden)

    Yu-shuang Feng

    2015-09-01

    Full Text Available In this research, a vermicular graphite cast iron brake drum was produced by cored wire injection in a one-step method. Silica sand and low-density alumina-silicate ceramic were used as molding materials in order to investigate the effect of cooling rate on percentage of vermicular graphite and mechanical properties of the brake drum casting. Several thermocouples were inserted into the casting in the desired positions to measure the temperature change. By means of one-step cored wire injection, the two residual concentrations of Mg and RE were effectively controlled in the ranges of 0.013%-0.017% and 0.019%-0.025%, respectively, which are crucial for the production of vermicular graphite cast iron and the formation of vermicular graphite. In addition, the cooling rate had a significant effect on the vermicular graphite percentage. In the case of the silica mold brake drum casting, there was an obvious difference in the cooling rate with the wall change, leading to a change in vermicular graphite percentage from 70.8% to 90%. In the low-density alumina-silicate ceramic mold casting, no obvious change in temperature was detected by the thermocouples and the percentage of the vermicular graphite was stable at 85%. Therefore, the vermicular graphite cast iron brake drum with a better combination of mechanical properties could be obtained.

  13. Removal of polycyclic aromatic hydrocarbons and phenols from coking wastewater by simultaneously synthesized organobentonite in a one-step process

    Institute of Scientific and Technical Information of China (English)

    Zhenhua Wu; Lizhong Zhu

    2012-01-01

    The optimal condition for a one-step process removing organic compounds from coiking wastewater by simultaneously synthesized organobentonite as a pretreatment was investigated.Results showed that sorption of organic compounds by organobentonite was positively correlated to the cation surfactant exchange on the bentonite and the octanol-water partition coefficient (Kow) of the solutes.With 0.75 g/L bentonite and 180 mg/L (60% of bentonite cation exchange capacity) cetyltrimethylammonium bromide,the removal efficiencies of the 16 polycyclic aromatic hydrocarbon (PAHs) specified by the US Environmental Protection Agency in coking waste0water except naphthalene were more than 90%,and that of benzo(a)pyrene was 99.5%.At the same time,the removal efficiencies of CODCr,NH3-N,volatile phenols,colour and turbidity were 28.6%,13.2%,8.9%,55% and 84.3%,respectively,and the ratio of BOD5/CODcr increased from 0.31 to 0.41.These results indicated that the one-step process had high removal efficiency for toxic and refractory hydrophobic organic compounds,and could improve the biodegradability of the coking wastewater.Therefore it could be a promising technology for the pretreatment of toxic and refractory organic wastewater.

  14. One-step synthesis and characterization of polyaniline nanofiber/silver nanoparticle composite networks as antibacterial agents.

    Science.gov (United States)

    Poyraz, Selcuk; Cerkez, Idris; Huang, Tung Shi; Liu, Zhen; Kang, Litao; Luo, Jujie; Zhang, Xinyu

    2014-11-26

    Through a facile and effective seeding polymerization reaction via a one-step redox/complexation process, which took place in aqueous medium at ambient temperature, silver nanoparticles (Ag NPs) embedded polyaniline nanofiber (PANI NF) networks were synthesized as antibacterial agents. During the reaction, not only NF morphology formation of the resulting conducting polymers (CPs) but also amplification of the aqueous silver nitrate (AgNO3) solutions' oxidative potentials were managed by vanadium pentoxide (V2O5) sol-gel nanofibers, which acted as well-known nanofibrous seeding agents and the auxiliary oxidative agent at the same time. The PANI/Ag nanocomposites were proven to exhibit excellent antibacterial property against both Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus. Antibacterial property performance and average life span of the nanocomposite network were optimized through the homogeneous distribution/embedment of Ag NPs within one-dimensional (1-D) PANI NF matrix. The antibacterial efficacy tests and nanocomposite material characterization results further indicated that the sole components of PANI/Ag have a synergistic effect to each other in terms of antibacterial property. Thus, this well-known catalytic seeding approach via a one-step oxidative polymerization reaction can be considered as a general methodology and a substantial fabrication tool to synthesize Ag NP decorated nanofibrillar PANI networks as advanced antibacterial agents.

  15. Humidity-Sensing Properties of One-Step Hydrothermally Synthesized Tin Dioxide-Decorated Graphene Nanocomposite on Polyimide Substrate

    Science.gov (United States)

    Zhang, Dongzhi; Chang, Hongyan; Liu, Runhua

    2016-08-01

    This paper demonstrates the one-step hydrothermal synthesis of a tin dioxide (SnO2)-decorated reduced graphene oxide (RGO) hybrid nanocomposite, which was drop-casted on a polyimide substrate as a humidity sensor. The as-synthesized hybrid was characterized in terms of its nanostructural, morphological and compositional features by SEM, XRD and nitrogen sorption. The humidity sensing properties of the presented RGO/SnO2 hybrid nanocomposite, such as repeatability, stability, response-recovery characteristics, were investigated by exposing it to a broad humidity range of 11-97% RH at room temperature. As a result, the sensor demonstrated a high sensitivity, a good repeatability, an acceptable linearity, a fast response/recovery characteristic and high long-term stability over a full humidity range measurement, indicating the unique advantages of one-step hydrothermal synthesis for sensor fabrication. The possible and proposed sensing mechanism for the sensor is mainly attributed to a humidity-induced transfer of charge carriers occuring at the interfaces and the swelling effect of RGO.

  16. Optimization of One-Step In Situ Transesterification Method for Accurate Quantification of EPA in Nannochloropsis gaditana

    Directory of Open Access Journals (Sweden)

    Yuting Tang

    2016-11-01

    Full Text Available Microalgae are a valuable source of lipid feedstocks for biodiesel and valuable omega-3 fatty acids. Nannochloropsis gaditana has emerged as a promising producer of eicosapentaenoic acid (EPA due to its fast growth rate and high EPA content. In the present study, the fatty acid profile of Nannochloropsis gaditana was found to be naturally high in EPA and devoid of docosahexaenoic acid (DHA, thereby providing an opportunity to maximize the efficacy of EPA production. Using an optimized one-step in situ transesterification method (methanol:biomass = 90 mL/g; HCl 5% by vol.; 70 °C; 1.5 h, the maximum fatty acid methyl ester (FAME yield of Nannochloropsis gaditana cultivated under rich condition was quantified as 10.04% ± 0.08% by weight with EPA-yields as high as 4.02% ± 0.17% based on dry biomass. The total FAME and EPA yields were 1.58- and 1.23-fold higher separately than that obtained using conventional two-step method (solvent system: methanol and chloroform. This one-step in situ method provides a fast and simple method to measure fatty acid methyl ester (FAME yields and could serve as a promising method to generate eicosapentaenoic acid methyl ester from microalgae.

  17. Effects of Sheet Resistance on mc-Si Selective Emitter Solar Cells Using Laser Opening and One-Step Diffusion

    Directory of Open Access Journals (Sweden)

    Sheng-Shih Wang

    2015-01-01

    Full Text Available In order to simplify process procedure and improve conversion efficiency (η, we present new steps of laser opening and one-step POCl3 diffusion to fabricate selective emitter (SE solar cells, in which heavily doped regions (HDR and lightly doped regions (LDR were formed simultaneously. For HDR, we divided six cells into two groups for POCl3 diffusion with sheet resistance (RS of 40 Ω/sq (for group A and 50 Ω/sq (for group B. The dry oxidation duration at a temperature of 850°C was 18, 25, and 35 min for the 3 different cells in each group. This created six SE samples with different RS pairings for the HDR and LDR. The optimal cell (sample SE2 with RS values of 40/81 Ω/Sq in HDR/LDR showed the best η of 16.20%, open circuit voltage (VOC of 612.52 mV, and fill factor (FF of 75.83%. The improvement ratios are 1.57% for η and 14.32% for external quantum efficiency (EQE as compared with those of the two-step diffusion process of our previous study. Moreover, the one-step laser opening process and omitting the step of removing the damage caused by laser ablation especially reduce chemistry pollution, thus showing ecofriendly process for use in industrial-scale production.

  18. Effect of moisture and drying time on the bond strength of the one-step self-etching adhesive system

    Directory of Open Access Journals (Sweden)

    Yoon Lee

    2012-08-01

    Full Text Available Objectives To investigate the effect of dentin moisture degree and air-drying time on dentin-bond strength of two different one-step self-etching adhesive systems. Materials and Methods Twenty-four human third molars were used for microtensile bond strength testing of G-Bond and Clearfil S3 Bond. The dentin surface was either blot-dried or air-dried before applying these adhesive agents. After application of the adhesive agent, three different air drying times were evaluated: 1, 5, and 10 sec. Composite resin was build up to 4 mm thickness and light cured for 40 sec with 2 separate layers. Then the tooth was sectioned and trimmed to measure the microtensile bond strength using a universal testing machine. The measured bond strengths were analyzed with three-way ANOVA and regression analysis was done (p = 0.05. Results All three factors, materials, dentin wetness and air drying time, showed significant effect on the microtensile bond strength. Clearfil S3 Bond, dry dentin surface and 10 sec air drying time showed higher bond strength. Conclusions Within the limitation of this experiment, air drying time after the application of the one-step self-etching adhesive agent was the most significant factor affecting the bond strength, followed by the material difference and dentin moisture before applying the adhesive agent.

  19. Properties of a Ni-FUSI gate formed by the EBV method and one-step RTA

    Science.gov (United States)

    Youwei, Zhang; Dawei, Xu; Li, Wan; Zhongjian, Wang; Chao, Xia; Xinhong, Cheng; Yuehui, Yu

    2012-03-01

    Nickel fully silicided (Ni-FUSI) gate material has been fabricated on a HfO2 surface to form a Ni-FUSI gate/HfO2/Si/Al (MIS) structure by using an ultra-high vacuum e-beam evaporation (EBV) method followed by a one step rapid thermal annealing (RTA) treatment. X-ray diffraction (XRD) and Raman spectroscopy were used to reveal the microstructures and electrical properties of the MIS structure. Results show that a one step post RTA treatment is enough to promote the full reaction of nickel silicide, compared with multiple RTA treatments. Furthermore, the HfO2 gate dielectric film is sensitive to heat treatment, and multiple RTA treatments can damage the electrical properties of the HfO2 film rather than improve them. By optimization of the sample fabrication technique, the MIS capacitor produces good high-frequency capacitance-voltage curves with a hysteresis of 30 mV, a work function of about 5.44-5.53 eV and leakage current density of only 1.45 × 10-8 A/cm2 at -1 V gate bias.

  20. Preparation of Fe3O4 magnetic fluid by one-step method with a microemulsion reactor

    Institute of Scientific and Technical Information of China (English)

    Zhang Wenjun; Zhang Huifang; Li Dezhong

    2006-01-01

    A W/O microemulsion was prepared with Span80-PS (petroleum sulfonate) as complex emulsifier,isopropanol as cosurfactant and kerosene as oil phase.The optimal constituents of microemulsion were found from pseudoternary phase diagrams:the mass ratio of Span80 to PS was 4:1 and complex surfactant to cosurfactant was 1:1.The Fe3O4 magnetic fluid was obtained by one-step method with the W/O microemulsion as mieroreactor to synthesize magnetic nanoparticles (reaction temperature was 30℃ and reaction time was 5 h) and kerosene as carder liquid.The magnetic fluid was investigated by TEM,XRD and fluorescence microscope.The magnetism was determined by Gouy magnetic balance,The average particle size of Fe3O4 was 7.4 nm,and magnetic particles were well-dispersed.The stable Fe3O4 magnetic fluid with good magnetism may be produced by one-step method in the W/O microemulsion.Accordingly,the traditional preparation method of magnetic fluid can be simplified greatly.