A simple and versatile design concept for fluorophore derivatives with intramolecular photostabilization

PubMed Central

van der Velde, Jasper H. M.; Oelerich, Jens; Huang, Jingyi; Smit, Jochem H.; Aminian Jazi, Atieh; Galiani, Silvia; Kolmakov, Kirill; Gouridis, Giorgos; Eggeling, Christian; Herrmann, Andreas; Roelfes, Gerard; Cordes, Thorben

2016-01-01

Intramolecular photostabilization via triple-state quenching was recently revived as a tool to impart synthetic organic fluorophores with ‘self-healing’ properties. To date, utilization of such fluorophore derivatives is rare due to their elaborate multi-step synthesis. Here we present a general strategy to covalently link a synthetic organic fluorophore simultaneously to a photostabilizer and biomolecular target via unnatural amino acids. The modular approach uses commercially available starting materials and simple chemical transformations. The resulting photostabilizer–dye conjugates are based on rhodamines, carbopyronines and cyanines with excellent photophysical properties, that is, high photostability and minimal signal fluctuations. Their versatile use is demonstrated by single-step labelling of DNA, antibodies and proteins, as well as applications in single-molecule and super-resolution fluorescence microscopy. We are convinced that the presented scaffolding strategy and the improved characteristics of the conjugates in applications will trigger the broader use of intramolecular photostabilization and help to emerge this approach as a new gold standard. PMID:26751640

Catalytic intermolecular carbon electrophile induced semipinacol rearrangement.

PubMed

Zhang, Qing-Wei; Zhang, Xiao-Bo; Li, Bao-Sheng; Xiang, Kai; Zhang, Fu-Min; Wang, Shao-Hua; Tu, Yong-Qiang

2013-02-25

A catalytic intermolecular carbon electrophile induced semipinacol rearrangement was realized and the asymmetric version was also preliminarily accomplished with 92% and 82% ee. The complex tricyclic system architecture with four continuous stereogenic centers could be achieved from simple starting materials in a single step under mild conditions.

Rapid one-step purification of single-cells encapsulated in alginate microcapsules from oil to aqueous phase using a hydrophobic filter paper: implications for single-cell experiments.

PubMed

Lee, Do-Hyun; Jang, Miran; Park, Je-Kyun

2014-10-01

By virtue of the biocompatibility and physical properties of hydrogel, picoliter-sized hydrogel microcapsules have been considered to be a biometric signature containing several features similar to that of encapsulated single cells, including phenotype, viability, and intracellular content. To maximize the experimental potential of encapsulating cells in hydrogel microcapsules, a method that enables efficient hydrogel microcapsule purification from oil is necessary. Current methods based on centrifugation for the conventional stepwise rinsing of oil, are slow and laborious and decrease the monodispersity and yield of the recovered hydrogel microcapsules. To remedy these shortcomings we have developed a simple one-step method to purify alginate microcapsules, containing a single live cell, from oil to aqueous phase. This method employs oil impregnation using a commercially available hydrophobic filter paper without multistep centrifugal purification and complicated microchannel networks. The oil-suspended alginate microcapsules encapsulating single cells from mammalian cancer cell lines (MCF-7, HepG2, and U937) and microorganisms (Chlorella vulgaris) were successfully exchanged to cell culture media by quick (~10 min) depletion of the surrounding oil phase without coalescence of neighboring microcapsules. Cell proliferation and high integrity of the microcapsules were also demonstrated by long-term incubation of microcapsules containing a single live cell. We expect that this method for the simple and rapid purification of encapsulated single-cell microcapsules will attain widespread adoption, assisting cell biologists and clinicians in the development of single-cell experiments. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

[Purification of arsenic-binding proteins in hamster plasma after oral administration of arsenite].

PubMed

Wang, Wenwen; Zhang, Min; Li, Chunhui; Qin, Yingjie; Hua, Naranmandura

2013-01-01

To purify the arsenic-binding proteins (As-BP) in hamster plasma after a single oral administration of arsenite (iAs(III)). Arsenite was given to hamsters in a single dose. Three types of HPLC columns, size exclusion, gel filtration and anion exchange columns, combined with an inductively coupled argon plasma mass spectrometer (ICP MS) were used to purify the As-BP in hamster plasma. SDS-PAGE was used to confirm the arsenic-binding proteins at each purification step. The three-step purification process successfully separated As-BP from other proteins (ie, arsenic unbound proteins) in hamster plasma. The molecular mass of purified As-BP in plasma was approximately 40-50 kD on SDS-PAGE. The three-step purification method is a simple and fast approach to purify the As-BP in plasma samples.

Photocleavable DNA Barcoding Antibodies for Multiplexed Protein Analysis in Single Cells.

PubMed

Ullal, Adeeti V; Weissleder, Ralph

2015-01-01

We describe a DNA-barcoded antibody sensing technique for single cell protein analysis in which the barcodes are photocleaved and digitally detected without amplification steps (Ullal et al., Sci Transl Med 6:219, 2014). After photocleaving the unique ~70 mer DNA barcodes we use a fluorescent hybridization technology for detection, similar to what is commonly done for nucleic acid readouts. This protocol offers a simple method for multiplexed protein detection using 100+ antibodies and can be performed on clinical samples as well as single cells.

Multistep hierarchical self-assembly of chiral nanopore arrays

PubMed Central

Kim, Hanim; Lee, Sunhee; Shin, Tae Joo; Korblova, Eva; Walba, David M.; Clark, Noel A.; Lee, Sang Bok; Yoon, Dong Ki

2014-01-01

A series of simple hierarchical self-assembly steps achieve self-organization from the centimeter to the subnanometer-length scales in the form of square-centimeter arrays of linear nanopores, each one having a single chiral helical nanofilament of large internal surface area and interfacial interactions based on chiral crystalline molecular arrangements. PMID:25246585

HPLC of the Polypeptides in a Hydrolyzate of Egg-White Lysozyme. An Experiment for the Undergraduate Biochemistry Laboratory.

ERIC Educational Resources Information Center

Richardson, W. S., III; Burns, L.

1988-01-01

Describes a simple high-performance liquid chromatography experiment for undergraduate biochemistry laboratories. The experiment illustrates the separation of polypeptides by a step gradient elution using a single pump instrument with no gradient attachments. Discusses instrumentation, analysis, a sample preparation, and results. (CW)

A simple, compact, and rigid piezoelectric step motor with large step size.

PubMed

Wang, Qi; Lu, Qingyou

2009-08-01

We present a novel piezoelectric stepper motor featuring high compactness, rigidity, simplicity, and any direction operability. Although tested in room temperature, it is believed to work in low temperatures, owing to its loose operation conditions and large step size. The motor is implemented with a piezoelectric scanner tube that is axially cut into almost two halves and clamp holds a hollow shaft inside at both ends via the spring parts of the shaft. Two driving voltages that singly deform the two halves of the piezotube in one direction and recover simultaneously will move the shaft in the opposite direction, and vice versa.

A simple, compact, and rigid piezoelectric step motor with large step size

NASA Astrophysics Data System (ADS)

Wang, Qi; Lu, Qingyou

2009-08-01

We present a novel piezoelectric stepper motor featuring high compactness, rigidity, simplicity, and any direction operability. Although tested in room temperature, it is believed to work in low temperatures, owing to its loose operation conditions and large step size. The motor is implemented with a piezoelectric scanner tube that is axially cut into almost two halves and clamp holds a hollow shaft inside at both ends via the spring parts of the shaft. Two driving voltages that singly deform the two halves of the piezotube in one direction and recover simultaneously will move the shaft in the opposite direction, and vice versa.

A Demo opto-electronic power source based on single-walled carbon nanotube sheets.

PubMed

Hu, Chunhua; Liu, Changhong; Chen, Luzhuo; Meng, Chuizhou; Fan, Shoushan

2010-08-24

It is known that single-walled carbon nanotubes (SWNTs) strongly absorb light, especially in the near-infrared (NIR) region, and convert it into heat. In fact, SWNTs also have considerable ability to convert heat into electricity. In this work, we show that SWNT sheets made from as-grown SWNT arrays display a large positive thermoelectric coefficient (p-type). We designed a simple SWNT device to convert illuminating NIR light directly into a notable voltage output, which was verified by experimental tests. Furthermore, by a simple functionalization step, the p- to n-type transition was conveniently achieved for the SWNT sheets. By integrating p- and n-type elements in series, we constructed a novel NIR opto-electronic power source, which outputs a large voltage that sums over the output of every single element. Additionally, the output of the demo device has shown a good linear relationship with NIR light power density, favorable for IR sensors.

DNA-Mediated Patterning of Single Quantum Dot Nanoarrays: A Reusable Platform for Single-Molecule Control

NASA Astrophysics Data System (ADS)

Huang, Da; Freeley, Mark; Palma, Matteo

2017-03-01

We present a facile strategy of general applicability for the assembly of individual nanoscale moieties in array configurations with single-molecule control. Combining the programming ability of DNA as a scaffolding material with a one-step lithographic process, we demonstrate the patterning of single quantum dots (QDs) at predefined locations on silicon and transparent glass surfaces: as proof of concept, clusters of either one, two, or three QDs were assembled in highly uniform arrays with a 60 nm interdot spacing within each cluster. Notably, the platform developed is reusable after a simple cleaning process and can be designed to exhibit different geometrical arrangements.

Single molecule targeted sequencing for cancer gene mutation detection.

PubMed

Gao, Yan; Deng, Liwei; Yan, Qin; Gao, Yongqian; Wu, Zengding; Cai, Jinsen; Ji, Daorui; Li, Gailing; Wu, Ping; Jin, Huan; Zhao, Luyang; Liu, Song; Ge, Liangjin; Deem, Michael W; He, Jiankui

2016-05-19

With the rapid decline in cost of sequencing, it is now affordable to examine multiple genes in a single disease-targeted clinical test using next generation sequencing. Current targeted sequencing methods require a separate step of targeted capture enrichment during sample preparation before sequencing. Although there are fast sample preparation methods available in market, the library preparation process is still relatively complicated for physicians to use routinely. Here, we introduced an amplification-free Single Molecule Targeted Sequencing (SMTS) technology, which combined targeted capture and sequencing in one step. We demonstrated that this technology can detect low-frequency mutations using artificially synthesized DNA sample. SMTS has several potential advantages, including simple sample preparation thus no biases and errors are introduced by PCR reaction. SMTS has the potential to be an easy and quick sequencing technology for clinical diagnosis such as cancer gene mutation detection, infectious disease detection, inherited condition screening and noninvasive prenatal diagnosis.

Multi-step rhodopsin inactivation schemes can account for the size variability of single photon responses in Limulus ventral photoreceptors

PubMed Central

1994-01-01

Limulus ventral photoreceptors generate highly variable responses to the absorption of single photons. We have obtained data on the size distribution of these responses, derived the distribution predicted from simple transduction cascade models and compared the theory and data. In the simplest of models, the active state of the visual pigment (defined by its ability to activate G protein) is turned off in a single reaction. The output of such a cascade is predicted to be highly variable, largely because of stochastic variation in the number of G proteins activated. The exact distribution predicted is exponential, but we find that an exponential does not adequately account for the data. The data agree much better with the predictions of a cascade model in which the active state of the visual pigment is turned off by a multi-step process. PMID:8057085

Monolithic carbon structures including suspended single nanowires and nanomeshes as a sensor platform

PubMed Central

2013-01-01

With the development of nanomaterial-based nanodevices, it became inevitable to develop cost-effective and simple nanofabrication technologies enabling the formation of nanomaterial assembly in a controllable manner. Herein, we present suspended monolithic carbon single nanowires and nanomeshes bridging two bulk carbon posts, fabricated in a designed manner using two successive UV exposure steps and a single pyrolysis step. The pyrolysis step is accompanied with a significant volume reduction, resulting in the shrinkage of micro-sized photoresist structures into nanoscale carbon structures. Even with the significant elongation of the suspended carbon nanowire induced by the volume reduction of the bulk carbon posts, the resultant tensional stress along the nanowire is not significant but grows along the wire thickness; this tensional stress gradient and the bent supports of the bridge-like carbon nanowire enhance structural robustness and alleviate the stiction problem that suspended nanostructures frequently experience. The feasibility of the suspended carbon nanostructures as a sensor platform was demonstrated by testing its electrochemical behavior, conductivity-temperature relationship, and hydrogen gas sensing capability. PMID:24256942

DNA combing on low-pressure oxygen plasma modified polysilsesquioxane substrates for single-molecule studies

PubMed Central

Sriram, K. K.; Chang, Chun-Ling; Rajesh Kumar, U.; Chou, Chia-Fu

2014-01-01

Molecular combing and flow-induced stretching are the most commonly used methods to immobilize and stretch DNA molecules. While both approaches require functionalization steps for the substrate surface and the molecules, conventionally the former does not take advantage of, as the latter, the versatility of microfluidics regarding robustness, buffer exchange capability, and molecule manipulation using external forces for single molecule studies. Here, we demonstrate a simple one-step combing process involving only low-pressure oxygen (O2) plasma modified polysilsesquioxane (PSQ) polymer layer to facilitate both room temperature microfluidic device bonding and immobilization of stretched single DNA molecules without molecular functionalization step. Atomic force microscopy and Kelvin probe force microscopy experiments revealed a significant increase in surface roughness and surface potential on low-pressure O2 plasma treated PSQ, in contrast to that with high-pressure O2 plasma treatment, which are proposed to be responsible for enabling effective DNA immobilization. We further demonstrate the use of our platform to observe DNA-RNA polymerase complexes and cancer drug cisplatin induced DNA condensation using wide-field fluorescence imaging. PMID:25332730

High-throughput purification of recombinant proteins using self-cleaving intein tags.

PubMed

Coolbaugh, M J; Shakalli Tang, M J; Wood, D W

2017-01-01

High throughput methods for recombinant protein production using E. coli typically involve the use of affinity tags for simple purification of the protein of interest. One drawback of these techniques is the occasional need for tag removal before study, which can be hard to predict. In this work, we demonstrate two high throughput purification methods for untagged protein targets based on simple and cost-effective self-cleaving intein tags. Two model proteins, E. coli beta-galactosidase (βGal) and superfolder green fluorescent protein (sfGFP), were purified using self-cleaving versions of the conventional chitin-binding domain (CBD) affinity tag and the nonchromatographic elastin-like-polypeptide (ELP) precipitation tag in a 96-well filter plate format. Initial tests with shake flask cultures confirmed that the intein purification scheme could be scaled down, with >90% pure product generated in a single step using both methods. The scheme was then validated in a high throughput expression platform using 24-well plate cultures followed by purification in 96-well plates. For both tags and with both target proteins, the purified product was consistently obtained in a single-step, with low well-to-well and plate-to-plate variability. This simple method thus allows the reproducible production of highly pure untagged recombinant proteins in a convenient microtiter plate format. Copyright © 2016 Elsevier Inc. All rights reserved.

Automated Discovery of Elementary Chemical Reaction Steps Using Freezing String and Berny Optimization Methods.

PubMed

Suleimanov, Yury V; Green, William H

2015-09-08

We present a simple protocol which allows fully automated discovery of elementary chemical reaction steps using in cooperation double- and single-ended transition-state optimization algorithms--the freezing string and Berny optimization methods, respectively. To demonstrate the utility of the proposed approach, the reactivity of several single-molecule systems of combustion and atmospheric chemistry importance is investigated. The proposed algorithm allowed us to detect without any human intervention not only "known" reaction pathways, manually detected in the previous studies, but also new, previously "unknown", reaction pathways which involve significant atom rearrangements. We believe that applying such a systematic approach to elementary reaction path finding will greatly accelerate the discovery of new chemistry and will lead to more accurate computer simulations of various chemical processes.

A facile single-step synthesis of polyvinylpyrrolidone-silver nanocomposites using a conventional spray dryer

NASA Astrophysics Data System (ADS)

Kim, Byung-Ho; Hyuck Kim, Yoon; Lee, Young Jin; Lee, Mi Jai; Kim, Jin-Ho; Hwang, Jonghee; Jeon, Dae-Woo

2018-01-01

We have developed a facile single-step synthesis of silver nanocomposite using a conventional spray dryer. We investigated the synthetic conditions by controlling the concentrations of the chemical reactants. Further, we confirmed the effect of the molecular weight of polyvinylpyrrolidones, and revealed that the molecular weight significantly affected the properties of the resultant silver nanocomposites. The long-term stability of the silver nanocomposites was tested, and little change was observed, even after storage for three months. Most of all, the simple commercial implementation, in combination with large-scale synthesis, possesses a variety of advantages, compared to conventional complicated and costly dry-process synthesis methods. Thus, our method presents opportunities for further investigation, for both lab-scale studies and large-scale industrial applications.

Application of Organophosphonic Acids by One-Step Supercritical CO2 on 1D and 2D Semiconductors: Toward Enhanced Electrical and Sensing Performances.

PubMed

Bhartia, Bhavesh; Bacher, Nadav; Jayaraman, Sundaramurthy; Khatib, Salam; Song, Jing; Guo, Shifeng; Troadec, Cedric; Puniredd, Sreenivasa Reddy; Srinivasan, Madapusi Palavedu; Haick, Hossam

2015-07-15

Formation of dense monolayers with proven atmospheric stability using simple fabrication conditions remains a major challenge for potential applications such as (bio)sensors, solar cells, surfaces for growth of biological cells, and molecular, organic, and plastic electronics. Here, we demonstrate a single-step modification of organophosphonic acids (OPA) on 1D and 2D structures using supercritical carbon dioxide (SCCO2) as a processing medium, with high stability and significantly shorter processing times than those obtained by the conventional physisorption-chemisorption method (2.5 h vs 48-60 h).The advantages of this approach in terms of stability and atmospheric resistivity are demonstrated on various 2D materials, such as indium-tin-oxide (ITO) and 2D Si surfaces. The advantage of the reported approach on electronic and sensing devices is demonstrated by Si nanowire field effect transistors (SiNW FETs), which have shown a few orders of magnitude higher electrical and sensing performances, compared with devices obtained by conventional approaches. The compatibility of the reported approach with various materials and its simple implementation with a single reactor makes it easily scalable for various applications.

A simple and novel method for RNA-seq library preparation of single cell cDNA analysis by hyperactive Tn5 transposase.

PubMed

Brouilette, Scott; Kuersten, Scott; Mein, Charles; Bozek, Monika; Terry, Anna; Dias, Kerith-Rae; Bhaw-Rosun, Leena; Shintani, Yasunori; Coppen, Steven; Ikebe, Chiho; Sawhney, Vinit; Campbell, Niall; Kaneko, Masahiro; Tano, Nobuko; Ishida, Hidekazu; Suzuki, Ken; Yashiro, Kenta

2012-10-01

Deep sequencing of single cell-derived cDNAs offers novel insights into oncogenesis and embryogenesis. However, traditional library preparation for RNA-seq analysis requires multiple steps with consequent sample loss and stochastic variation at each step significantly affecting output. Thus, a simpler and better protocol is desirable. The recently developed hyperactive Tn5-mediated library preparation, which brings high quality libraries, is likely one of the solutions. Here, we tested the applicability of hyperactive Tn5-mediated library preparation to deep sequencing of single cell cDNA, optimized the protocol, and compared it with the conventional method based on sonication. This new technique does not require any expensive or special equipment, which secures wider availability. A library was constructed from only 100 ng of cDNA, which enables the saving of precious specimens. Only a few steps of robust enzymatic reaction resulted in saved time, enabling more specimens to be prepared at once, and with a more reproducible size distribution among the different specimens. The obtained RNA-seq results were comparable to the conventional method. Thus, this Tn5-mediated preparation is applicable for anyone who aims to carry out deep sequencing for single cell cDNAs. Copyright © 2012 Wiley Periodicals, Inc.

Accelerating Drug Development: Antiviral Therapies for Emerging Viruses as a Model.

PubMed

Everts, Maaike; Cihlar, Tomas; Bostwick, J Robert; Whitley, Richard J

2017-01-06

Drug discovery and development is a lengthy and expensive process. Although no one, simple, single solution can significantly accelerate this process, steps can be taken to avoid unnecessary delays. Using the development of antiviral therapies as a model, we describe options for acceleration that cover target selection, assay development and high-throughput screening, hit confirmation, lead identification and development, animal model evaluations, toxicity studies, regulatory issues, and the general drug discovery and development infrastructure. Together, these steps could result in accelerated timelines for bringing antiviral therapies to market so they can treat emerging infections and reduce human suffering.

Efficient production of erythroid, megakaryocytic and myeloid cells, using single cell-derived iPSC colony differentiation.

PubMed

Hansen, Marten; Varga, Eszter; Aarts, Cathelijn; Wust, Tatjana; Kuijpers, Taco; von Lindern, Marieke; van den Akker, Emile

2018-04-28

Hematopoietic differentiation of human induced pluripotent stem cells (iPSCs) provide opportunities not only for fundamental research and disease modelling/drug testing but also for large-scale production of blood effector cells for future clinical application. Although there are multiple ways to differentiate human iPSCs towards hematopoietic lineages, there is a need to develop reproducible and robust protocols. Here we introduce an efficient way to produce three major blood cell types using a standardized differentiation protocol that starts with a single hematopoietic initiation step. This system is feeder-free, avoids EB-formation, starts with a hematopoietic initiation step based on a novel single cell-derived iPSC colony differentiation and produces multi-potential progenitors within 8-10 days. Followed by lineage-specific growth factor supplementation these cells can be matured into well characterized erythroid, megakaryocytic and myeloid cells with high-purity, without transcription factor overexpression or any kind of pre-purification step. This standardized differentiation system provides a simple platform to produce specific blood cells in a reproducible manner for hematopoietic development studies, disease modelling, drug testing and the potential for future therapeutic applications. Copyright © 2018. Published by Elsevier B.V.

One-step fabrication of 3D silver paste electrodes into microfluidic devices for enhanced droplet-based cell sorting

NASA Astrophysics Data System (ADS)

Rao, Lang; Cai, Bo; Yu, Xiao-Lei; Guo, Shi-Shang; Liu, Wei; Zhao, Xing-Zhong

2015-05-01

3D microelectrodes are one-step fabricated into a microfluidic droplet separator by filling conductive silver paste into PDMS microchambers. The advantages of 3D silver paste electrodes in promoting droplet sorting accuracy are systematically demonstrated by theoretical calculation, numerical simulation and experimental validation. The employment of 3D electrodes also helps to decrease the droplet sorting voltage, guaranteeing that cells encapsulated in droplets undergo chip-based sorting processes are at better metabolic status for further potential cellular assays. At last, target droplet containing single cell are selectively sorted out from others by an appropriate electric pulse. This method provides a simple and inexpensive alternative to fabricate 3D electrodes, and it is expected our 3D electrode-integrated microfluidic droplet separator platform can be widely used in single cell operation and analysis.

LiCoPO4 cathode from a CoHPO4·xH2O nanoplate precursor for high voltage Li-ion batteries

DOE Office of Scientific and Technical Information (OSTI.GOV)

Choi, Daiwon; Li, Xiaolin; Henderson, Wesley A.

2016-02-01

Highly crystalline LiCoPO4/C cathode has been synthesized without any impurities via single step solid-state reaction using CoHPO4xH2O nanoplates as a precursor obtained by simple precipitation route. The electrochemical test shows specific capacity as high as 125mAh/g at charge/discharge rate of C/10. Synthesis approach for obtaining CoHPO4xH2O nanoplate precursor and final LiCoPO4/C cathode using single step solid-state reaction have been characterized using X-ray diffraction, thermos gravimetric analyses (TGA) – differential scanning calorimetry (DSC), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). The electrochemical test and cycling stability using different electrolytes, additive and separator have been investigated.

Bacterial Expression of a Single-Chain Variable Fragment (scFv) Antibody against Ganoderic Acid A: A Cost-Effective Approach for Quantitative Analysis Using the scFv-Based Enzyme-Linked Immunosorbent Assay.

PubMed

Yusakul, Gorawit; Nuntawong, Poomraphie; Sakamoto, Seiichi; Ratnatilaka Na Bhuket, Pahweenvaj; Kohno, Toshitaka; Kikkawa, Nao; Rojsitthisak, Pornchai; Shimizu, Kuniyoshi; Tanaka, Hiroyuki; Morimoto, Satoshi

2017-01-01

Due to the highly specific binding between an antibody and its target, superior analytical performances was obtained by immunoassays for phytochemical analysis over conventional chromatographic techniques. Here, we describe a simple method for producing a functional single-chain variable fragment (scFv) antibody against ganoderic acid A (GAA), a pharmacologically active metabolite from Ganoderma lingzhi. The Escherichia coli BL21(DE3) strain produced a large amount of anti-GAA scFv. However, in vitro refolding steps, which partially recovered the reactivity of the scFv, were required. Interestingly, the functional scFv was expressed as a soluble and active form in the cytoplasm of an engineered E. coli SHuffle ® strain. Purified anti-GAA scFv, which yielded 2.56 mg from 1 L of culture medium, was obtained from simple and inexpensive procedures for expression and purification. The anti-GAA scFv-based indirect competitive enzyme-linked immunosorbent assay (icELISA) exhibited high sensitivity (linearity: 0.078-1.25 µg/mL) with precision (CV: ≤6.20%) and reliability (recovery: 100.1-101.8%) for GAA determination. In summary, the approach described here is an inexpensive, simple, and efficient expression system that extends the application of anti-GAA scFv-based immunoassays. In addition, when in vitro refolding steps can be skipped, the cost and complexity of scFv antibody production can be minimized.

In-depth analysis and characterization of a dual damascene process with respect to different CD

NASA Astrophysics Data System (ADS)

Krause, Gerd; Hofmann, Detlef; Habets, Boris; Buhl, Stefan; Gutsch, Manuela; Lopez-Gomez, Alberto; Kim, Wan-Soo; Thrun, Xaver

2018-03-01

In a 200 mm high volume environment, we studied data from a dual damascene process. Dual damascene is a combination of lithography, etch and CMP that is used to create copper lines and contacts in one single step. During these process steps, different metal CD are measured by different measurement methods. In this study, we analyze the key numbers of the different measurements after different process steps and develop simple models to predict the electrical behavior* . In addition, radial profiles have been analyzed of both inline measurement parameters and electrical parameters. A matching method was developed based on inline and electrical data. Finally, correlation analysis for radial signatures is presented that can be used to predict excursions in electrical signatures.

Optimization of single-base-pair mismatch discrimination in oligonucleotide microarrays

NASA Technical Reports Server (NTRS)

Urakawa, Hidetoshi; El Fantroussi, Said; Smidt, Hauke; Smoot, James C.; Tribou, Erik H.; Kelly, John J.; Noble, Peter A.; Stahl, David A.

2003-01-01

The discrimination between perfect-match and single-base-pair-mismatched nucleic acid duplexes was investigated by using oligonucleotide DNA microarrays and nonequilibrium dissociation rates (melting profiles). DNA and RNA versions of two synthetic targets corresponding to the 16S rRNA sequences of Staphylococcus epidermidis (38 nucleotides) and Nitrosomonas eutropha (39 nucleotides) were hybridized to perfect-match probes (18-mer and 19-mer) and to a set of probes having all possible single-base-pair mismatches. The melting profiles of all probe-target duplexes were determined in parallel by using an imposed temperature step gradient. We derived an optimum wash temperature for each probe and target by using a simple formula to calculate a discrimination index for each temperature of the step gradient. This optimum corresponded to the output of an independent analysis using a customized neural network program. These results together provide an experimental and analytical framework for optimizing mismatch discrimination among all probes on a DNA microarray.

A simple and rapid method for high-resolution visualization of single-ion tracks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Omichi, Masaaki; Center for Collaborative Research, Anan National College of Technology, Anan, Tokushima 774-0017; Choi, Wookjin

2014-11-15

Prompt determination of spatial points of single-ion tracks plays a key role in high-energy particle induced-cancer therapy and gene/plant mutations. In this study, a simple method for the high-resolution visualization of single-ion tracks without etching was developed through the use of polyacrylic acid (PAA)-N, N’-methylene bisacrylamide (MBAAm) blend films. One of the steps of the proposed method includes exposure of the irradiated films to water vapor for several minutes. Water vapor was found to promote the cross-linking reaction of PAA and MBAAm to form a bulky cross-linked structure; the ion-track scars were detectable at a nanometer scale by atomic forcemore » microscopy. This study demonstrated that each scar is easily distinguishable, and the amount of generated radicals of the ion tracks can be estimated by measuring the height of the scars, even in highly dense ion tracks. This method is suitable for the visualization of the penumbra region in a single-ion track with a high spatial resolution of 50 nm, which is sufficiently small to confirm that a single ion hits a cell nucleus with a size ranging between 5 and 20 μm.« less

Adaptation in Tunably Rugged Fitness Landscapes: The Rough Mount Fuji Model

PubMed Central

Neidhart, Johannes; Szendro, Ivan G.; Krug, Joachim

2014-01-01

Much of the current theory of adaptation is based on Gillespie’s mutational landscape model (MLM), which assumes that the fitness values of genotypes linked by single mutational steps are independent random variables. On the other hand, a growing body of empirical evidence shows that real fitness landscapes, while possessing a considerable amount of ruggedness, are smoother than predicted by the MLM. In the present article we propose and analyze a simple fitness landscape model with tunable ruggedness based on the rough Mount Fuji (RMF) model originally introduced by Aita et al. in the context of protein evolution. We provide a comprehensive collection of results pertaining to the topographical structure of RMF landscapes, including explicit formulas for the expected number of local fitness maxima, the location of the global peak, and the fitness correlation function. The statistics of single and multiple adaptive steps on the RMF landscape are explored mainly through simulations, and the results are compared to the known behavior in the MLM model. Finally, we show that the RMF model can explain the large number of second-step mutations observed on a highly fit first-step background in a recent evolution experiment with a microvirid bacteriophage. PMID:25123507

Firing patterns in the adaptive exponential integrate-and-fire model.

PubMed

Naud, Richard; Marcille, Nicolas; Clopath, Claudia; Gerstner, Wulfram

2008-11-01

For simulations of large spiking neuron networks, an accurate, simple and versatile single-neuron modeling framework is required. Here we explore the versatility of a simple two-equation model: the adaptive exponential integrate-and-fire neuron. We show that this model generates multiple firing patterns depending on the choice of parameter values, and present a phase diagram describing the transition from one firing type to another. We give an analytical criterion to distinguish between continuous adaption, initial bursting, regular bursting and two types of tonic spiking. Also, we report that the deterministic model is capable of producing irregular spiking when stimulated with constant current, indicating low-dimensional chaos. Lastly, the simple model is fitted to real experiments of cortical neurons under step current stimulation. The results provide support for the suitability of simple models such as the adaptive exponential integrate-and-fire neuron for large network simulations.

A single step reversed-phase high performance liquid chromatography separation of polar and non-polar lipids.

PubMed

Olsson, Petter; Holmbäck, Jan; Herslöf, Bengt

2014-11-21

This paper reports a simple chromatographic system to separate lipids classes as well as their molecular species. By the use of phenyl coated silica as stationary phase in combination with a simple mobile phase consisting of methanol and water, all tested lipid classes elute within 30 min. Furthermore, a method to accurately predict retention times of specific lipid components for this type of chromatography is presented. Common detection systems were used, namely evaporative light scattering detection (ELSD), charged aerosol detection (CAD), electrospray mass spectrometry (ESI-MS), and UV detection. Copyright © 2014 Elsevier B.V. All rights reserved.

Synthesis of partially and fully fused polyaromatics by annulative chlorophenylene dimerization.

PubMed

Koga, Yoshito; Kaneda, Takeshi; Saito, Yutaro; Murakami, Kei; Itami, Kenichiro

2018-01-26

Since the discovery by Ullmann and Bielecki in 1901, reductive dimerization (or homocoupling) of aryl halides has been extensively exploited for the generation of a range of biaryl-based functional molecules. In contrast to the single-point connection in these products, edge-sharing fused aromatic systems have not generally been accessible from simple aryl halides via annulation cascades. Here we report a single-step synthesis of fused aromatics with a triphenylene core by the palladium-catalyzed annulative dimerization of structurally and functionally diverse chlorophenylenes through double carbon-hydrogen bond activation. The partially fused polyaromatics can be transformed into fully fused, small graphene nanoribbons, which are otherwise difficult to synthesize. This simple, yet powerful, method allows access to functional π-systems of interest in optoelectronics research. Copyright © 2018, The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

Three-dimensional photonic crystals created by single-step multi-directional plasma etching.

PubMed

Suzuki, Katsuyoshi; Kitano, Keisuke; Ishizaki, Kenji; Noda, Susumu

2014-07-14

We fabricate 3D photonic nanostructures by simultaneous multi-directional plasma etching. This simple and flexible method is enabled by controlling the ion-sheath in reactive-ion-etching equipment. We realize 3D photonic crystals on single-crystalline silicon wafers and show high reflectance (>95%) and low transmittance (<-15dB) at optical communication wavelengths, suggesting the formation of a complete photonic bandgap. Moreover, our method simply demonstrates Si-based 3D photonic crystals that show the photonic bandgap effect in a shorter wavelength range around 0.6 μm, where further fine structures are required.

High quality factor whispering gallery modes from self-assembled hexagonal GaN rods grown by metal-organic vapor phase epitaxy.

PubMed

Tessarek, C; Sarau, G; Kiometzis, M; Christiansen, S

2013-02-11

Self-assembled GaN rods were grown on sapphire by metal-organic vapor phase epitaxy using a simple two-step method that relies first on a nitridation step followed by GaN epitaxy. The mask-free rods formed without any additional catalyst. Most of the vertically aligned rods exhibit a regular hexagonal shape with sharp edges and smooth sidewall facets. Cathodo- and microphotoluminescence investigations were carried out on single GaN rods. Whispering gallery modes with quality factors greater than 4000 were measured demonstrating the high morphological and optical quality of the self-assembled GaN rods.

A Simple Method to Simultaneously Detect and Identify Spikes from Raw Extracellular Recordings.

PubMed

Petrantonakis, Panagiotis C; Poirazi, Panayiota

2015-01-01

The ability to track when and which neurons fire in the vicinity of an electrode, in an efficient and reliable manner can revolutionize the neuroscience field. The current bottleneck lies in spike sorting algorithms; existing methods for detecting and discriminating the activity of multiple neurons rely on inefficient, multi-step processing of extracellular recordings. In this work, we show that a single-step processing of raw (unfiltered) extracellular signals is sufficient for both the detection and identification of active neurons, thus greatly simplifying and optimizing the spike sorting approach. The efficiency and reliability of our method is demonstrated in both real and simulated data.

Computationally Aided Absolute Stereochemical Determination of Enantioenriched Amines.

PubMed

Zhang, Jun; Gholami, Hadi; Ding, Xinliang; Chun, Minji; Vasileiou, Chrysoula; Nehira, Tatsuo; Borhan, Babak

2017-03-17

A simple and efficient protocol for sensing the absolute stereochemistry and enantiomeric excess of chiral monoamines is reported. Preparation of the sample requires a single-step reaction of the 1,1'-(bromomethylene)dinaphthalene (BDN) with the chiral amine. Analysis of the exciton coupled circular dichroism generated from the BDN-derivatized chiral amine sample, along with comparison to conformational analysis performed computationally, yields the absolute stereochemistry of the parent chiral monoamine.

Plasmonic SERS nanochips and nanoprobes for medical diagnostics and bio-energy applications

NASA Astrophysics Data System (ADS)

Ngo, Hoan T.; Wang, Hsin-Neng; Crawford, Bridget M.; Fales, Andrew M.; Vo-Dinh, Tuan

2017-02-01

The development of rapid, easy-to-use, cost-effective, high accuracy, and high sensitive DNA detection methods for molecular diagnostics has been receiving increasing interest. Over the last five years, our laboratory has developed several chip-based DNA detection techniques including the molecular sentinel-on-chip (MSC), the multiplex MSC, and the inverse molecular sentinel-on-chip (iMS-on-Chip). In these techniques, plasmonic surface-enhanced Raman scattering (SERS) Nanowave chips were functionalized with DNA probes for single-step DNA detection. Sensing mechanisms were based on hybridization of target sequences and DNA probes, resulting in a distance change between SERS reporters and the Nanowave chip's gold surface. This distance change resulted in change in SERS intensity, thus indicating the presence and capture of the target sequences. Our techniques were single-step DNA detection techniques. Target sequences were detected by simple delivery of sample solutions onto DNA probe-functionalized Nanowave chips and SERS signals were measured after 1h - 2h incubation. Target sequence labeling or washing to remove unreacted components was not required, making the techniques simple, easy-to-use, and cost effective. The usefulness of the techniques for medical diagnostics was illustrated by the detection of genetic biomarkers for respiratory viral infection and of dengue virus 4 DNA.

Variable tunneling barriers in FEBID based PtC metal-matrix nanocomposites as a transducing element for humidity sensing.

PubMed

Kolb, Florian; Schmoltner, Kerstin; Huth, Michael; Hohenau, Andreas; Krenn, Joachim; Klug, Andreas; List, Emil J W; Plank, Harald

2013-08-02

The development of simple gas sensing concepts is still of great interest for science and technology. The demands on an ideal device would be a single-step fabrication method providing a device which is sensitive, analyte-selective, quantitative, and reversible without special operating/reformation conditions such as high temperatures or special environments. In this study we demonstrate a new gas sensing concept based on a nanosized PtC metal-matrix system fabricated in a single step via focused electron beam induced deposition (FEBID). The sensors react selectively on polar H2O molecules quantitatively and reversibly without any special reformation conditions after detection events, whereas non-polar species (O2, CO2, N2) produce no response. The key elements are isolated Pt nanograins (2-3 nm) which are embedded in a dielectric carbon matrix. The electrical transport in such materials is based on tunneling effects in the correlated variable range hopping regime, where the dielectric carbon matrix screens the electric field between the particles, which governs the final conductivity. The specific change of these dielectric properties by the physisorption of polar gas molecules (H2O) can change the tunneling probability and thus the overall conductivity, allowing their application as a simple and straightforward sensing concept.

Interfacial complexation in microfluidic droplets for single-step fabrication of microcapsule

NASA Astrophysics Data System (ADS)

Kaufman, Gilad; Nejati, Siamak; Sarfati, Raphael; Boltyanskiy, Rostislav; Williams, Danielle; Liu, Wei; Schloss, Ashley; Regan, Lynn; Yan, Elsa; Dufrense, Eric; Loewenberg, Michael; Osuji, Chinedum

We present microfluidic interfacial complexation in emulsion droplets as a simple single-step approach for fabricating a large variety of stable monodisperse microcapsules with tailored mechanical properties, protein binding and controlled release behavior. We rely on electrostatic interactions and hydrogen bonding to direct the assembly of complementary species at oil-water droplet interfaces to form microcapsules with polyelectrolyte shells, composite polyelectrolyte-nanoparticle shells, and copolymer-nanofiber shells. Additionally, we demonstrate the formation of microcapsules by adsorption of an amphiphilic bacterial hydrophobin, BslA, at oil-in-water and water-in-oil droplets, and protein capture on these capsules using engineered variants of the hydrophobin. We discuss the composition dependence of mechanical properties, shell thickness and release behavior, and regimes of stability for microcapsule fabrication. Nanoparticle based microcapsules display an intriguing plastic deformation response which enables the formation of large aspect ratio asperities by pipette aspiration of the shell.

High-Resolution “Fleezers”: Dual-Trap Optical Tweezers Combined with Single-Molecule Fluorescence Detection

PubMed Central

Whitley, Kevin D.; Comstock, Matthew J.; Chemla, Yann R.

2017-01-01

Recent advances in optical tweezers have greatly expanded their measurement capabilities. A new generation of hybrid instrument that combines nanomechanical manipulation with fluorescence detection—fluorescence optical tweezers, or “fleezers”—is providing a powerful approach to study complex macromolecular dynamics. Here, we describe a combined high-resolution optical trap/confocal fluorescence microscope that can simultaneously detect sub-nanometer displacements, sub-piconewton forces, and single-molecule fluorescence signals. The primary technical challenge to these hybrid instruments is how to combine both measurement modalities without sacrificing the sensitivity of either one. We present general design principles to overcome this challenge and provide detailed, step-by-step instructions to implement them in the construction and alignment of the instrument. Lastly, we present a set of protocols to perform a simple, proof-of-principle experiment that highlights the instrument capabilities. PMID:27844430

Using multifield measurements to eliminate alignment degeneracies in the JWST testbed telescope

NASA Astrophysics Data System (ADS)

Sabatke, Erin; Acton, Scott; Schwenker, John; Towell, Tim; Carey, Larkin; Shields, Duncan; Contos, Adam; Leviton, Doug

2007-09-01

The primary mirror of the James Webb Space Telescope (JWST) consists of 18 segments and is 6.6 meters in diameter. A sequence of commissioning steps is carried out at a single field point to align the segments. At that single field point, though, the segmented primary mirror can compensate for aberrations caused by misalignments of the remaining mirrors. The misalignments can be detected in the wavefronts of off-axis field points. The Multifield (MF) step in the commissioning process surveys five field points and uses a simple matrix multiplication to calculate corrected positions for the secondary and primary mirrors. A demonstration of the Multifield process was carried out on the JWST Testbed Telescope (TBT). The results show that the Multifield algorithm is capable of reducing the field dependency of the TBT to about 20 nm RMS, relative to the TBT design nominal field dependency.

Improving Signal-to-Noise Ratio in Scanning Transmission Electron Microscopy Energy-Dispersive X-Ray (STEM-EDX) Spectrum Images Using Single-Atomic-Column Cross-Correlation Averaging.

PubMed

Jeong, Jong Seok; Mkhoyan, K Andre

2016-06-01

Acquiring an atomic-resolution compositional map of crystalline specimens has become routine practice, thus opening possibilities for extracting subatomic information from such maps. A key challenge for achieving subatomic precision is the improvement of signal-to-noise ratio (SNR) of compositional maps. Here, we report a simple and reliable solution for achieving high-SNR energy-dispersive X-ray (EDX) spectroscopy spectrum images for individual atomic columns. The method is based on standard cross-correlation aided by averaging of single-column EDX maps with modifications in the reference image. It produces EDX maps with minimal specimen drift, beam drift, and scan distortions. Step-by-step procedures to determine a self-consistent reference map with a discussion on the reliability, stability, and limitations of the method are presented here.

Integrin Beta 1 Suppresses Multilayering of a Simple Epithelium

PubMed Central

Chen, Jichao; Krasnow, Mark A.

2012-01-01

Epithelia are classified as either simple, a single cell layer thick, or stratified (multilayered). Stratified epithelia arise from simple epithelia during development, and transcription factor p63 functions as a key positive regulator of epidermal stratification. Here we show that deletion of integrin beta 1 (Itgb1) in the developing mouse airway epithelium abrogates airway branching and converts this monolayer epithelium into a multilayer epithelium with more than 10 extra layers. Mutant lung epithelial cells change mitotic spindle orientation to seed outer layers, and cells in different layers become molecularly and functionally distinct, hallmarks of normal stratification. However, mutant lung epithelial cells do not activate p63 and do not switch to the stratified keratin profile of epidermal cells. These data, together with previous data implicating Itgb1 in regulation of epidermal stratification, suggest that the simple-versus-stratified developmental decision may involve not only stratification inducers like p63 but suppressors like Itgb1 that prevent simple epithelia from inappropriately activating key steps in the stratification program. PMID:23285215

Numerical calculation on a two-step subdiffusion behavior of lateral protein movement in plasma membranes

NASA Astrophysics Data System (ADS)

Sumi, Tomonari; Okumoto, Atsushi; Goto, Hitoshi; Sekino, Hideo

2017-10-01

A two-step subdiffusion behavior of lateral movement of transmembrane proteins in plasma membranes has been observed by using single-molecule experiments. A nested double-compartment model where large compartments are divided into several smaller ones has been proposed in order to explain this observation. These compartments are considered to be delimited by membrane-skeleton "fences" and membrane-protein "pickets" bound to the fences. We perform numerical simulations of a master equation using a simple two-dimensional lattice model to investigate the heterogeneous diffusion dynamics behavior of transmembrane proteins within plasma membranes. We show that the experimentally observed two-step subdiffusion process can be described using fence and picket models combined with decreased local diffusivity of transmembrane proteins in the vicinity of the pickets. This allows us to explain the two-step subdiffusion behavior without explicitly introducing nested double compartments.

Hierarchical tone mapping for high dynamic range image visualization

NASA Astrophysics Data System (ADS)

Qiu, Guoping; Duan, Jiang

2005-07-01

In this paper, we present a computationally efficient, practically easy to use tone mapping techniques for the visualization of high dynamic range (HDR) images in low dynamic range (LDR) reproduction devices. The new method, termed hierarchical nonlinear linear (HNL) tone-mapping operator maps the pixels in two hierarchical steps. The first step allocates appropriate numbers of LDR display levels to different HDR intensity intervals according to the pixel densities of the intervals. The second step linearly maps the HDR intensity intervals to theirs allocated LDR display levels. In the developed HNL scheme, the assignment of LDR display levels to HDR intensity intervals is controlled by a very simple and flexible formula with a single adjustable parameter. We also show that our new operators can be used for the effective enhancement of ordinary images.

Gold-coated polydimethylsiloxane microwells for high-throughput electrochemiluminescence analysis of intracellular glucose at single cells.

PubMed

Xia, Juan; Zhou, Junyu; Zhang, Ronggui; Jiang, Dechen; Jiang, Depeng

2018-06-04

In this communication, a gold-coated polydimethylsiloxane (PDMS) chip with cell-sized microwells was prepared through a stamping and spraying process that was applied directly for high-throughput electrochemiluminescence (ECL) analysis of intracellular glucose at single cells. As compared with the previous multiple-step fabrication of photoresist-based microwells on the electrode, the preparation process is simple and offers fresh electrode surface for higher luminescence intensity. More luminescence intensity was recorded from cell-retained microwells than that at the planar region among the microwells that was correlated with the content of intracellular glucose. The successful monitoring of intracellular glucose at single cells using this PDMS chip will provide an alternative strategy for high-throughput single-cell analysis. Graphical abstract ᅟ.

Combating omission errors through task analysis and good reminders.

PubMed

Reason, J

2002-03-01

Leaving out necessary task steps is the single most common human error type. Certain task steps possess characteristics that are more likely to provoke omissions than others, and can be identified in advance. The paper reports two studies. The first, involving a simple photocopier, established that failing to remove the last page of the original is the commonest omission. This step possesses four distinct error-provoking features that combine their effects in an additive fashion. The second study examined the degree to which everyday memory aids satisfy five features of a good reminder: conspicuity, contiguity, content, context, and countability. A close correspondence was found between the percentage use of strategies and the degree to which they satisfied these five criteria. A three stage omission management programme was outlined: task analysis (identifying discrete task steps) of some safety critical activity; assessing the omission likelihood of each step; and the choice and application of a suitable reminder. Such a programme is applicable to a variety of healthcare procedures.

Comparison of two PCR-based methods and automated DNA sequencing for prop-1 genotyping in Ames dwarf mice.

PubMed

Gerstner, Arpad; DeFord, James H; Papaconstantinou, John

2003-07-25

Ames dwarfism is caused by a homozygous single nucleotide mutation in the pituitary specific prop-1 gene, resulting in combined pituitary hormone deficiency, reduced growth and extended lifespan. Thus, these mice serve as an important model system for endocrinological, aging and longevity studies. Because the phenotype of wild type and heterozygous mice is undistinguishable, it is imperative for successful breeding to accurately genotype these animals. Here we report a novel, yet simple, approach for prop-1 genotyping using PCR-based allele-specific amplification (PCR-ASA). We also compare this method to other potential genotyping techniques, i.e. PCR-based restriction fragment length polymorphism analysis (PCR-RFLP) and fluorescence automated DNA sequencing. We demonstrate that the single-step PCR-ASA has several advantages over the classical PCR-RFLP because the procedure is simple, less expensive and rapid. To further increase the specificity and sensitivity of the PCR-ASA, we introduced a single-base mismatch at the 3' penultimate position of the mutant primer. Our results also reveal that the fluorescence automated DNA sequencing has limitations for detecting a single nucleotide polymorphism in the prop-1 gene, particularly in heterozygotes.

Multi-Model Combination techniques for Hydrological Forecasting: Application to Distributed Model Intercomparison Project Results

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ajami, N K; Duan, Q; Gao, X

2005-04-11

This paper examines several multi-model combination techniques: the Simple Multi-model Average (SMA), the Multi-Model Super Ensemble (MMSE), Modified Multi-Model Super Ensemble (M3SE) and the Weighted Average Method (WAM). These model combination techniques were evaluated using the results from the Distributed Model Intercomparison Project (DMIP), an international project sponsored by the National Weather Service (NWS) Office of Hydrologic Development (OHD). All of the multi-model combination results were obtained using uncalibrated DMIP model outputs and were compared against the best uncalibrated as well as the best calibrated individual model results. The purpose of this study is to understand how different combination techniquesmore » affect the skill levels of the multi-model predictions. This study revealed that the multi-model predictions obtained from uncalibrated single model predictions are generally better than any single member model predictions, even the best calibrated single model predictions. Furthermore, more sophisticated multi-model combination techniques that incorporated bias correction steps work better than simple multi-model average predictions or multi-model predictions without bias correction.« less

A simple test of choice stepping reaction time for assessing fall risk in people with multiple sclerosis.

PubMed

Tijsma, Mylou; Vister, Eva; Hoang, Phu; Lord, Stephen R

2017-03-01

Purpose To determine (a) the discriminant validity for established fall risk factors and (b) the predictive validity for falls of a simple test of choice stepping reaction time (CSRT) in people with multiple sclerosis (MS). Method People with MS (n = 210, 21-74y) performed the CSRT, sensorimotor, balance and neuropsychological tests in a single session. They were then followed up for falls using monthly fall diaries for 6 months. Results The CSRT test had excellent discriminant validity with respect to established fall risk factors. Frequent fallers (≥3 falls) performed significantly worse in the CSRT test than non-frequent fallers (0-2 falls). With the odds of suffering frequent falls increasing 69% with each SD increase in CSRT (OR = 1.69, 95% CI: 1.27-2.26, p = <0.001). In regression analysis, CSRT was best explained by sway, time to complete the 9-Hole Peg test, knee extension strength of the weaker leg, proprioception and the time to complete the Trails B test (multiple R 2   =   0.449, p < 0.001). Conclusions A simple low tech CSRT test has excellent discriminative and predictive validity in relation to falls in people with MS. This test may prove useful in documenting longitudinal changes in fall risk in relation to MS disease progression and effects of interventions. Implications for rehabilitation Good choice stepping reaction time (CSRT) is required for maintaining balance. A simple low-tech CSRT test has excellent discriminative and predictive validity in relation to falls in people with MS. This test may prove useful documenting longitudinal changes in fall risk in relation to MS disease progression and effects of interventions.

Asynchronous collision integrators: Explicit treatment of unilateral contact with friction and nodal restraints

PubMed Central

Wolff, Sebastian; Bucher, Christian

2013-01-01

This article presents asynchronous collision integrators and a simple asynchronous method treating nodal restraints. Asynchronous discretizations allow individual time step sizes for each spatial region, improving the efficiency of explicit time stepping for finite element meshes with heterogeneous element sizes. The article first introduces asynchronous variational integration being expressed by drift and kick operators. Linear nodal restraint conditions are solved by a simple projection of the forces that is shown to be equivalent to RATTLE. Unilateral contact is solved by an asynchronous variant of decomposition contact response. Therein, velocities are modified avoiding penetrations. Although decomposition contact response is solving a large system of linear equations (being critical for the numerical efficiency of explicit time stepping schemes) and is needing special treatment regarding overconstraint and linear dependency of the contact constraints (for example from double-sided node-to-surface contact or self-contact), the asynchronous strategy handles these situations efficiently and robust. Only a single constraint involving a very small number of degrees of freedom is considered at once leading to a very efficient solution. The treatment of friction is exemplified for the Coulomb model. Special care needs the contact of nodes that are subject to restraints. Together with the aforementioned projection for restraints, a novel efficient solution scheme can be presented. The collision integrator does not influence the critical time step. Hence, the time step can be chosen independently from the underlying time-stepping scheme. The time step may be fixed or time-adaptive. New demands on global collision detection are discussed exemplified by position codes and node-to-segment integration. Numerical examples illustrate convergence and efficiency of the new contact algorithm. Copyright © 2013 The Authors. International Journal for Numerical Methods in Engineering published by John Wiley & Sons, Ltd. PMID:23970806

Single-shot digital holography by use of the fractional Talbot effect.

PubMed

Martínez-León, Lluís; Araiza-E, María; Javidi, Bahram; Andrés, Pedro; Climent, Vicent; Lancis, Jesús; Tajahuerce, Enrique

2009-07-20

We present a method for recording in-line single-shot digital holograms based on the fractional Talbot effect. In our system, an image sensor records the interference between the light field scattered by the object and a properly codified parallel reference beam. A simple binary two-dimensional periodic grating is used to codify the reference beam generating a periodic three-step phase distribution over the sensor plane by fractional Talbot effect. This provides a method to perform single-shot phase-shifting interferometry at frame rates only limited by the sensor capabilities. Our technique is well adapted for dynamic wavefront sensing applications. Images of the object are digitally reconstructed from the digital hologram. Both computer simulations and experimental results are presented.

Purification of Proteins Fused to Maltose-Binding Protein.

PubMed

Lebendiker, Mario; Danieli, Tsafi

2017-01-01

Maltose-Binding Protein (MBP) is one of the most popular fusion partners being used for producing recombinant proteins in bacterial cells. MBP allows the use of a simple capture affinity step on Amylose-Agarose or Dextrin-Sepharose columns, resulting in a protein that is often 70-90 % pure in a single step. In addition to protein isolation applications, MBP provides a high degree of translation, and facilitates the proper folding and solubility of the target protein. This paper describes efficient procedures for isolating highly purified MBP target proteins. Special attention is given to considerations for downstream applications such as structural determination studies, protein activity assays, and assessing the chemical characteristics of the target protein.

Migraines: Simple Steps to Head Off the Pain

MedlinePlus

Migraines: Simple steps to head off the pain Migraines cause pain as real as the pain of ... Healthy habits and simple nonmedical remedies sometimes stop migraines before they start. By Mayo Clinic Staff Medication ...

Compatibility of pedigree-based and marker-based relationship matrices for single-step genetic evaluation.

PubMed

Christensen, Ole F

2012-12-03

Single-step methods provide a coherent and conceptually simple approach to incorporate genomic information into genetic evaluations. An issue with single-step methods is compatibility between the marker-based relationship matrix for genotyped animals and the pedigree-based relationship matrix. Therefore, it is necessary to adjust the marker-based relationship matrix to the pedigree-based relationship matrix. Moreover, with data from routine evaluations, this adjustment should in principle be based on both observed marker genotypes and observed phenotypes, but until now this has been overlooked. In this paper, I propose a new method to address this issue by 1) adjusting the pedigree-based relationship matrix to be compatible with the marker-based relationship matrix instead of the reverse and 2) extending the single-step genetic evaluation using a joint likelihood of observed phenotypes and observed marker genotypes. The performance of this method is then evaluated using two simulated datasets. The method derived here is a single-step method in which the marker-based relationship matrix is constructed assuming all allele frequencies equal to 0.5 and the pedigree-based relationship matrix is constructed using the unusual assumption that animals in the base population are related and inbred with a relationship coefficient γ and an inbreeding coefficient γ / 2. Taken together, this γ parameter and a parameter that scales the marker-based relationship matrix can handle the issue of compatibility between marker-based and pedigree-based relationship matrices. The full log-likelihood function used for parameter inference contains two terms. The first term is the REML-log-likelihood for the phenotypes conditional on the observed marker genotypes, whereas the second term is the log-likelihood for the observed marker genotypes. Analyses of the two simulated datasets with this new method showed that 1) the parameters involved in adjusting marker-based and pedigree-based relationship matrices can depend on both observed phenotypes and observed marker genotypes and 2) a strong association between these two parameters exists. Finally, this method performed at least as well as a method based on adjusting the marker-based relationship matrix. Using the full log-likelihood and adjusting the pedigree-based relationship matrix to be compatible with the marker-based relationship matrix provides a new and interesting approach to handle the issue of compatibility between the two matrices in single-step genetic evaluation.

Molgenis-impute: imputation pipeline in a box.

PubMed

Kanterakis, Alexandros; Deelen, Patrick; van Dijk, Freerk; Byelas, Heorhiy; Dijkstra, Martijn; Swertz, Morris A

2015-08-19

Genotype imputation is an important procedure in current genomic analysis such as genome-wide association studies, meta-analyses and fine mapping. Although high quality tools are available that perform the steps of this process, considerable effort and expertise is required to set up and run a best practice imputation pipeline, particularly for larger genotype datasets, where imputation has to scale out in parallel on computer clusters. Here we present MOLGENIS-impute, an 'imputation in a box' solution that seamlessly and transparently automates the set up and running of all the steps of the imputation process. These steps include genome build liftover (liftovering), genotype phasing with SHAPEIT2, quality control, sample and chromosomal chunking/merging, and imputation with IMPUTE2. MOLGENIS-impute builds on MOLGENIS-compute, a simple pipeline management platform for submission and monitoring of bioinformatics tasks in High Performance Computing (HPC) environments like local/cloud servers, clusters and grids. All the required tools, data and scripts are downloaded and installed in a single step. Researchers with diverse backgrounds and expertise have tested MOLGENIS-impute on different locations and imputed over 30,000 samples so far using the 1,000 Genomes Project and new Genome of the Netherlands data as the imputation reference. The tests have been performed on PBS/SGE clusters, cloud VMs and in a grid HPC environment. MOLGENIS-impute gives priority to the ease of setting up, configuring and running an imputation. It has minimal dependencies and wraps the pipeline in a simple command line interface, without sacrificing flexibility to adapt or limiting the options of underlying imputation tools. It does not require knowledge of a workflow system or programming, and is targeted at researchers who just want to apply best practices in imputation via simple commands. It is built on the MOLGENIS compute workflow framework to enable customization with additional computational steps or it can be included in other bioinformatics pipelines. It is available as open source from: https://github.com/molgenis/molgenis-imputation.

Efficient assembly of polysubstituted pyrroles via a (3 + 2) cycloaddition/skeletal rearrangement/redox isomerization cascade reaction.

PubMed

Yu, Yuanyuan; Wang, Chunyu; He, Xinze; Yao, Xiaotong; Zu, Liansuo

2014-07-03

An unprecedented cascade strategy, used in conjunction with a redox isomerization, for the synthesis of 3-allyl pyrroles is reported. In a single step, readily accessible simple starting materials are transformed into highly substituted pyrroles with high efficiency. The products obtained contain allyl substituents, which can be readily elaborated to other useful functional groups. The reaction proceeds through an unusual (3 + 2) cycloaddition/skeletal rearrangement/redox isomerization pathway.

Wire-packed heat exchangers for dilution refrigerators.

PubMed

Polturak, E; Rappaport, M; Rosenbaum, R

1978-03-01

Very simple wire-packed step heat exchangers for dilution refrigerators are described. No sintering is used in fabrication. Flow impedances and thermal resistance between the liquid and the copper wires are low. A refrigerator with five wire-packed heat exchangers in addition to a countercurrent heat exchanger attains a temperature of 11.4 mK with a single mixing chamber and 6.1 mK with two mixing chambers. High cooling power is achieved at modest (3)He circulation rates.

A simple and effective process for noise reduction of multichannel cortical field potential recordings in freely moving rats.

PubMed

Shaw, Fu-Zen; Yen, Chen Tung; Chen, Ruei Feng

2003-04-15

Simple and useful steps, i.e. placing a grounded plate under the recording chamber as well as using multiple reference electrodes, are introduced here for obtaining reliable low-noise recordings of brain activity in freely moving rats. A general circuit model was built to analyze the electrical interference of both single-grounded and two-reference ground-free recording configurations. In both simulated and realistic conditions under two recording states, 60-Hz magnitude was in the microvolt range. Moreover, the noise was significantly reduced by shortening the distance between the subject and the grounded plate under the recording chamber. Furthermore, in chronically implanted rats, average 60-Hz interference of multichannel electroencephalograms of two-reference ground-free recordings (3.74 +/- 0.18 microV) was significantly smaller than that of the single-grounded condition (9.03 +/- 1.98 microV). Thus, we demonstrated that a lower-noise recording can be achieved by a two-reference configuration and a closely-placed metal grounded plate in an open-field circumstance. As compared to the use of a Faraday cage, this simple procedure is of benefit for long-term behavioral tracking with a video camera and for pharmacological experiments.

Application of the θ-method to a telegraphic model of fluid flow in a dual-porosity medium

NASA Astrophysics Data System (ADS)

González-Calderón, Alfredo; Vivas-Cruz, Luis X.; Herrera-Hernández, Erik César

2018-01-01

This work focuses mainly on the study of numerical solutions, which are obtained using the θ-method, of a generalized Warren and Root model that includes a second-order wave-like equation in its formulation. The solutions approximately describe the single-phase hydraulic head in fractures by considering the finite velocity of propagation by means of a Cattaneo-like equation. The corresponding discretized model is obtained by utilizing a non-uniform grid and a non-uniform time step. A simple relationship is proposed to give the time-step distribution. Convergence is analyzed by comparing results from explicit, fully implicit, and Crank-Nicolson schemes with exact solutions: a telegraphic model of fluid flow in a single-porosity reservoir with relaxation dynamics, the Warren and Root model, and our studied model, which is solved with the inverse Laplace transform. We find that the flux and the hydraulic head have spurious oscillations that most often appear in small-time solutions but are attenuated as the solution time progresses. Furthermore, we show that the finite difference method is unable to reproduce the exact flux at time zero. Obtaining results for oilfield production times, which are in the order of months in real units, is only feasible using parallel implicit schemes. In addition, we propose simple parallel algorithms for the memory flux and for the explicit scheme.

Temperature-programmed natural convection for micromixing and biochemical reaction in a single microfluidic chamber.

PubMed

Kim, Sung-Jin; Wang, Fang; Burns, Mark A; Kurabayashi, Katsuo

2009-06-01

Micromixing is a crucial step for biochemical reactions in microfluidic networks. A critical challenge is that the system containing micromixers needs numerous pumps, chambers, and channels not only for the micromixing but also for the biochemical reactions and detections. Thus, a simple and compatible design of the micromixer element for the system is essential. Here, we propose a simple, yet effective, scheme that enables micromixing and a biochemical reaction in a single microfluidic chamber without using any pumps. We accomplish this process by using natural convection in conjunction with alternating heating of two heaters for efficient micromixing, and by regulating capillarity for sample transport. As a model application, we demonstrate micromixing and subsequent polymerase chain reaction (PCR) for an influenza viral DNA fragment. This process is achieved in a platform of a microfluidic cartridge and a microfabricated heating-instrument with a fast thermal response. Our results will significantly simplify micromixing and a subsequent biochemical reaction that involves reagent heating in microfluidic networks.

Silicon nanowires: where mechanics and optics meet at the nanoscale.

PubMed

Ramos, Daniel; Gil-Santos, Eduardo; Malvar, Oscar; Llorens, Jose M; Pini, Valerio; San Paulo, Alvaro; Calleja, Montserrat; Tamayo, Javier

2013-12-06

Mechanical transducers based on nanowires promise revolutionary advances in biological sensing and force microscopy/spectroscopy. A crucial step is the development of simple and non-invasive techniques able to detect displacements with subpicometer sensitivity per unit bandwidth. Here, we design suspended tapered silicon nanowires supporting a range of optical resonances that confine and efficiently scatter light in the visible range. Then, we develop an optical method for efficiently coupling the evanescent field to the regular interference pattern generated by an incoming laser beam and the reflected beam from the substrate underneath the nanowire. This optomechanical coupling is here applied to measure the displacement of 50 nm wide nanowires with sensitivity on the verge of 1 fm/Hz(1/2) at room temperature with a simple laser interferometry set-up. This method opens the door to the measurement of the Brownian motion of ultrashort nanowires for the detection of single biomolecular recognition events in liquids, and single molecule spectroscopy in vacuum.

A colorimetric probe based on desensitized ionene-stabilized gold nanoparticles for single-step test for sulfate ions.

PubMed

Arkhipova, Viktoriya V; Apyari, Vladimir V; Dmitrienko, Stanislava G

2015-03-15

Desensitized ionene-stabilized gold nanoparticles have been prepared and applied as a colorimetric probe for the single-step test for sulfate ions at the relatively high concentration level. The approach is based on aggregation of the nanoparticles leading to the change in absorption spectra and color of the solution. These nanoparticles are characterized by the decreased sensitivity due to both electrostatic and steric stabilization, which allows for simple, and rapid direct single-step determination of sulfate at the relatively high concentration level in real water samples without sample pretreatment or dilution. Influence of different factors (the time of interaction, pH, the concentrations of sulfate ions and the nanoparticles) on the aggregation and analytical performance of the procedure was investigated. The method allows for the determination of sulfate ions in the mass range of 0.2-0.4 mg with RSD of 5% from the sample volume of less than 2 mL. It has a sharp dependence of the colorimetric response on the concentration of sulfate, which makes it prospective for indicating deviations of the sulfate concentration regarding some declared value chosen within the above range. The time of the analysis is 2 min. The method was applied to the analysis of mineral water samples. Copyright © 2014 Elsevier B.V. All rights reserved.

Three-dimensional Aerographite-GaN hybrid networks: Single step fabrication of porous and mechanically flexible materials for multifunctional applications

PubMed Central

Schuchardt, Arnim; Braniste, Tudor; Mishra, Yogendra K.; Deng, Mao; Mecklenburg, Matthias; Stevens-Kalceff, Marion A.; Raevschi, Simion; Schulte, Karl; Kienle, Lorenz; Adelung, Rainer; Tiginyanu, Ion

2015-01-01

Three dimensional (3D) elastic hybrid networks built from interconnected nano- and microstructure building units, in the form of semiconducting-carbonaceous materials, are potential candidates for advanced technological applications. However, fabrication of these 3D hybrid networks by simple and versatile methods is a challenging task due to the involvement of complex and multiple synthesis processes. In this paper, we demonstrate the growth of Aerographite-GaN 3D hybrid networks using ultralight and extremely porous carbon based Aerographite material as templates by a single step hydride vapor phase epitaxy process. The GaN nano- and microstructures grow on the surface of Aerographite tubes and follow the network architecture of the Aerographite template without agglomeration. The synthesized 3D networks are integrated with the properties from both, i.e., nanoscale GaN structures and Aerographite in the form of flexible and semiconducting composites which could be exploited as next generation materials for electronic, photonic, and sensors applications. PMID:25744694

Origins of multicellular evolvability in snowflake yeast

PubMed Central

Ratcliff, William C.; Fankhauser, Johnathon D.; Rogers, David W.; Greig, Duncan; Travisano, Michael

2015-01-01

Complex life has arisen through a series of ‘major transitions’ in which collectives of formerly autonomous individuals evolve into a single, integrated organism. A key step in this process is the origin of higher-level evolvability, but little is known about how higher-level entities originate and gain the capacity to evolve as an individual. Here we report a single mutation that not only creates a new level of biological organization, but also potentiates higher-level evolvability. Disrupting the transcription factor ACE2 in Saccharomyces cerevisiae prevents mother–daughter cell separation, generating multicellular ‘snowflake’ yeast. Snowflake yeast develop through deterministic rules that produce geometrically defined clusters that preclude genetic conflict and display a high broad-sense heritability for multicellular traits; as a result they are preadapted to multicellular adaptation. This work demonstrates that simple microevolutionary changes can have profound macroevolutionary consequences, and suggests that the formation of clonally developing clusters may often be the first step to multicellularity. PMID:25600558

Advanced metal lift-off process using electron-beam flood exposure of single-layer photoresist

NASA Astrophysics Data System (ADS)

Minter, Jason P.; Ross, Matthew F.; Livesay, William R.; Wong, Selmer S.; Narcy, Mark E.; Marlowe, Trey

1999-06-01

In the manufacture of many types of integrated circuit and thin film devices, it is desirable to use a lift-of process for the metallization step to avoid manufacturing problems encountered when creating metal interconnect structures using plasma etch. These problems include both metal adhesion and plasma etch difficulties. Key to the success of the lift-off process is the creation of a retrograde or undercut profile in the photoresists before the metal deposition step. Until now, lift-off processing has relied on costly multi-layer photoresists schemes, image reversal, and non-repeatable photoresist processes to obtain the desired lift-off profiles in patterned photoresist. This paper present a simple, repeatable process for creating robust, user-defined lift-off profiles in single layer photoresist using a non-thermal electron beam flood exposure. For this investigation, lift-off profiles created using electron beam flood exposure of many popular photoresists were evaluated. Results of lift-off profiles created in positive tone AZ7209 and ip3250 are presented here.

Multidomain proteins under force

NASA Astrophysics Data System (ADS)

Valle-Orero, Jessica; Andrés Rivas-Pardo, Jaime; Popa, Ionel

2017-04-01

Advancements in single-molecule force spectroscopy techniques such as atomic force microscopy and magnetic tweezers allow investigation of how domain folding under force can play a physiological role. Combining these techniques with protein engineering and HaloTag covalent attachment, we investigate similarities and differences between four model proteins: I10 and I91—two immunoglobulin-like domains from the muscle protein titin, and two α + β fold proteins—ubiquitin and protein L. These proteins show a different mechanical response and have unique extensions under force. Remarkably, when normalized to their contour length, the size of the unfolding and refolding steps as a function of force reduces to a single master curve. This curve can be described using standard models of polymer elasticity, explaining the entropic nature of the measured steps. We further validate our measurements with a simple energy landscape model, which combines protein folding with polymer physics and accounts for the complex nature of tandem domains under force. This model can become a useful tool to help in deciphering the complexity of multidomain proteins operating under force.

A Minimal Optical Trapping and Imaging Microscopy System

PubMed Central

Hernández Candia, Carmen Noemí; Tafoya Martínez, Sara; Gutiérrez-Medina, Braulio

2013-01-01

We report the construction and testing of a simple and versatile optical trapping apparatus, suitable for visualizing individual microtubules (∼25 nm in diameter) and performing single-molecule studies, using a minimal set of components. This design is based on a conventional, inverted microscope, operating under plain bright field illumination. A single laser beam enables standard optical trapping and the measurement of molecular displacements and forces, whereas digital image processing affords real-time sample visualization with reduced noise and enhanced contrast. We have tested our trapping and imaging instrument by measuring the persistence length of individual double-stranded DNA molecules, and by following the stepping of single kinesin motor proteins along clearly imaged microtubules. The approach presented here provides a straightforward alternative for studies of biomaterials and individual biomolecules. PMID:23451216

Self-digitization microfluidic chip for absolute quantification of mRNA in single cells.

PubMed

Thompson, Alison M; Gansen, Alexander; Paguirigan, Amy L; Kreutz, Jason E; Radich, Jerald P; Chiu, Daniel T

2014-12-16

Quantification of mRNA in single cells provides direct insight into how intercellular heterogeneity plays a role in disease progression and outcomes. Quantitative polymerase chain reaction (qPCR), the current gold standard for evaluating gene expression, is insufficient for providing absolute measurement of single-cell mRNA transcript abundance. Challenges include difficulties in handling small sample volumes and the high variability in measurements. Microfluidic digital PCR provides far better sensitivity for minute quantities of genetic material, but the typical format of this assay does not allow for counting of the absolute number of mRNA transcripts samples taken from single cells. Furthermore, a large fraction of the sample is often lost during sample handling in microfluidic digital PCR. Here, we report the absolute quantification of single-cell mRNA transcripts by digital, one-step reverse transcription PCR in a simple microfluidic array device called the self-digitization (SD) chip. By performing the reverse transcription step in digitized volumes, we find that the assay exhibits a linear signal across a wide range of total RNA concentrations and agrees well with standard curve qPCR. The SD chip is found to digitize a high percentage (86.7%) of the sample for single-cell experiments. Moreover, quantification of transferrin receptor mRNA in single cells agrees well with single-molecule fluorescence in situ hybridization experiments. The SD platform for absolute quantification of single-cell mRNA can be optimized for other genes and may be useful as an independent control method for the validation of mRNA quantification techniques.

Development of a Launch Vehicle Manufacturing Process. Chapter 4

NASA Technical Reports Server (NTRS)

Vickers, John; Munafo, Paul M. (Technical Monitor)

2002-01-01

One of the goals of this chapter is to provide sufficient information so that you can develop a manufacturing process for a potential launch vehicle. With the variety of manufacturing options available, you might ask how this can possibly be done in the span of a single chapter. Actually, it will be quite simple because a basic manufacturing process is nothing more than a set of logical steps that are iterated until they produce a desired product. Although these statements seem simple and logical, don't let this simplicity fool you. Manufacturing problems with launch vehicles and their subassemblies have been the primary cause of project failures because the vehicle concept delivered to the manufacturing floor could not be built as designed.

Biomechanical influences on balance recovery by stepping.

PubMed

Hsiao, E T; Robinovitch, S N

1999-10-01

Stepping represents a common means for balance recovery after a perturbation to upright posture. Yet little is known regarding the biomechanical factors which determine whether a step succeeds in preventing a fall. In the present study, we developed a simple pendulum-spring model of balance recovery by stepping, and used this to assess how step length and step contact time influence the effort (leg contact force) and feasibility of balance recovery by stepping. We then compared model predictions of step characteristics which minimize leg contact force to experimentally observed values over a range of perturbation strengths. At all perturbation levels, experimentally observed step execution times were higher than optimal, and step lengths were smaller than optimal. However, the predicted increase in leg contact force associated with these deviations was substantial only for large perturbations. Furthermore, increases in the strength of the perturbation caused subjects to take larger, quicker steps, which reduced their predicted leg contact force. We interpret these data to reflect young subjects' desire to minimize recovery effort, subject to neuromuscular constraints on step execution time and step length. Finally, our model predicts that successful balance recovery by stepping is governed by a coupling between step length, step execution time, and leg strength, so that the feasibility of balance recovery decreases unless declines in one capacity are offset by enhancements in the others. This suggests that one's risk for falls may be affected more by small but diffuse neuromuscular impairments than by larger impairment in a single motor capacity.

Single mode low-NA step index Yb-doped fiber design for output powers beyond 4kW (Conference Presentation)

NASA Astrophysics Data System (ADS)

Beier, Franz; Proske, Fritz; Hupel, Christian; Kuhn, Stefan; Hein, Sigrun; Sattler, Bettina; Nold, Johannes; Haarlammert, Nicoletta; Schreiber, Thomas; Eberhardt, Ramona; Tünnermann, Andreas

2017-03-01

Fiber amplifiers are representing one of the most promising solid state laser concepts, due to the compact setup size, a simple thermal management and furthermore excellent beam quality. In this contribution, we report on the latest results from a low-NA, large mode area single mode fiber with a single mode output power beyond 4 kW without any indication of mode instabilities or nonlinear effects and high slope efficiency. Furthermore, we quantify the influence of the bending diameter of our manufactured low NA fiber on the average core loss by an OFDR measurement and determine the optimal bending diameter in comparison to a second fiber with a slightly changed NA. The fibers used in the experiments were fabricated by MCVD technology combined with the solution doping technique. The investigation indicates the limitation of the step index fiber design and its influence on the use in high power fiber amplifiers. We demonstrate, that even a slightly change in the core NA crucially influences the minimum bending diameter of the fiber and has to be taken into account in applications. The measured output power represents to the best of our knowledge the highest single mode output power of an amplifier fiber ever reported on.

On-Chip Magnetic Platform for Single-Particle Manipulation with Integrated Electrical Feedback.

PubMed

Monticelli, Marco; Torti, Andrea; Cantoni, Matteo; Petti, Daniela; Albisetti, Edoardo; Manzin, Alessandra; Guerriero, Erica; Sordan, Roman; Gervasoni, Giacomo; Carminati, Marco; Ferrari, Giorgio; Sampietro, Marco; Bertacco, Riccardo

2016-02-17

Methods for the manipulation of single magnetic particles have become very interesting, in particular for in vitro biological studies. Most of these studies require an external microscope to provide the operator with feedback for controlling the particle motion, thus preventing the use of magnetic particles in high-throughput experiments. In this paper, a simple and compact system with integrated electrical feedback is presented, implementing in the very same device both the manipulation and detection of the transit of single particles. The proposed platform is based on zig-zag shaped magnetic nanostructures, where transverse magnetic domain walls are pinned at the corners and attract magnetic particles in suspension. By applying suitable external magnetic fields, the domain walls move to the nearest corner, thus causing the step by step displacement of the particles along the nanostructure. The very same structure is also employed for detecting the bead transit. Indeed, the presence of the magnetic particle in suspension over the domain wall affects the depinning field required for its displacement. This characteristic field can be monitored through anisotropic magnetoresistance measurements, thus implementing an integrated electrical feedback of the bead transit. In particular, the individual manipulation and detection of single 1-μm sized beads is demonstrated. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Engine-Level Simulation of Liquid Rocket Combustion Instabilities: Transcritical Combustion Simulations in Single Injector Configurations

DTIC Science & Technology

2012-03-01

simple 1-step mechanism taking into account 4 species: CH4, O2, CO2 and H2O. Figure 2. Multiblock grid for the CVRC experiment. Left: Overall view, Right... Supercritical (and subcritical) fluid behavior and modeling: drops, streams, shear and mixing layers, jets and sprays. Progress in Energy and...hydrogen shear-coaxial jet flames at supercritical pressure. Com- bustion science and technology, 178(1-3):229–252, 2006. 12 B. E. Poling, J. M. Prausnitz

Copper-catalyzed domino cycloaddition/C-N coupling/cyclization/(C-H arylation): an efficient three-component synthesis of nitrogen polyheterocycles.

PubMed

Qian, Wenyuan; Wang, Hao; Allen, Jennifer

2013-10-11

A cat of all trades: A single copper catalyst promoted up to three reaction steps with separate catalytic cycles in a domino sequence (azide-alkyne cycloaddition/Goldberg amidation/Camps cyclization/(CH arylation)) for the rapid construction of complex heterocycles from three simple components under mild conditions. Facile cleavage of the triazole ring enables further elaboration of the condensation products. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Single-crystalline nanogap electrodes: enhancing the nanowire-breakdown process with a gaseous environment.

PubMed

Suga, Hiroshi; Sumiya, Touru; Furuta, Shigeo; Ueki, Ryuichi; Miyazawa, Yosuke; Nishijima, Takuya; Fujita, Jun-ichi; Tsukagoshi, Kazuhito; Shimizu, Tetsuo; Naitoh, Yasuhisa

2012-10-24

A method for fabricating single-crystalline nanogaps on Si substrates was developed. Polycrystalline Pt nanowires on Si substrates were broken down by current flow under various gaseous environments. The crystal structure of the nanogap electrode was evaluated using scanning electron microscopy and transmission electron microscopy. Nanogap electrodes sandwiched between Pt-large-crystal-grains were obtained by the breakdown of the wire in an O(2) or H(2) atmosphere. These nanogap electrodes show intense spots in the electron diffraction pattern. The diffraction pattern corresponds to Pt (111), indicating that single-crystal grains are grown by the electrical wire breakdown process in an O(2) or H(2) atmosphere. The Pt wires that have (111)-texture and coherent boundaries can be considered ideal as interconnectors for single molecular electronics. The simple method for fabrication of a single-crystalline nanogap is one of the first steps toward standard nanogap electrodes for single molecular instruments and opens the door to future research on physical phenomena in nanospaces.

A pelvic motion driven electrical stimulator for drop-foot treatment.

PubMed

Chen, Shih-Wei; Chen, Shih-Ching; Chen, Chiun-Fan; Lai, Jin-Shin; Kuo, Te-Son

2009-01-01

Foot switches operating with force sensitive resistors placed in the shoe sole were considered as an effective way for driving FES assisted walking systems in gait restoration. However, the reliability and durability of the foot switches run down after a certain number of steps. As an alternative for foot switches, a simple, portable, and easy to handle motion driven electrical stimulator (ES) is provided for drop foot treatment. The device is equipped with a single tri-axis accelerometer worn on the pelvis, a commercial dual channel electrical stimulator, and a controller unit. By monitoring the pelvic rotation and acceleration during a walking cycle, the events including heel strike and toe off of each step is thereby predicted by a post-processing neural network model.

Vectorial strain gauge method using single flexible orthogonal polydimethylsiloxane gratings

NASA Astrophysics Data System (ADS)

Guo, Hao; Tang, Jun; Qian, Kun; Tsoukalas, Dimitris; Zhao, Miaomiao; Yang, Jiangtao; Zhang, Binzhen; Chou, Xiujian; Liu, Jun; Xue, Chenyang; Zhang, Wendong

2016-03-01

A vectorial strain gauge method using a single sensing element is reported based on the double-sided polydimethylsiloxane (PDMS) Fraunhofer diffraction gratings structures. Using O2 plasma treatment steps, orthogonal wrinkled gratings were fabricated on both sides of a pre-strained PDMS film. Diffracted laser spots from this structure have been used to experimentally demonstrate, that any applied strain can be quantitatively characterized in both the x and y directions with an error of less than 0.6% and with a gauge factor of approximately 10. This simple and low cost technology which is completely different from the traditional vectorial strain gauge method, can be applied to surface vectorial strain measurement and multi-axis integrated mechanical sensors.

Software manual for operating particle displacement tracking data acquisition and reduction system

NASA Technical Reports Server (NTRS)

Wernet, Mark P.

1991-01-01

The software manual is presented. The necessary steps required to record, analyze, and reduce Particle Image Velocimetry (PIV) data using the Particle Displacement Tracking (PDT) technique are described. The new PDT system is an all electronic technique employing a CCD video camera and a large memory buffer frame-grabber board to record low velocity (less than or equal to 20 cm/s) flows. Using a simple encoding scheme, a time sequence of single exposure images are time coded into a single image and then processed to track particle displacements and determine 2-D velocity vectors. All the PDT data acquisition, analysis, and data reduction software is written to run on an 80386 PC.

Experimental quantum information processing with the Talbot effect

NASA Astrophysics Data System (ADS)

Sawada, K.; Walborn, S. P.

2018-07-01

We report a proof of concept experiment illustrating the implementation of several simple quantum logic gates on D-level quantum systems (quDits) using the Talbot effect. A number of QuDit states are encoded into the transverse profile of a paraxial laser beam using a spatial light modulator. These states are transformed through a diagonal phase element and then free-propagation via the fractional Talbot effect, demonstrating the realization of some well-known single quDit gates in quantum computation. Our classical optics experiment allows us to identify several important technical details, and serves as a first experimental step in performing D-dimensional quantum operations with single photons or other quantum systems using this scheme.

Bayesian Analysis of Evolutionary Divergence with Genomic Data under Diverse Demographic Models.

PubMed

Chung, Yujin; Hey, Jody

2017-06-01

We present a new Bayesian method for estimating demographic and phylogenetic history using population genomic data. Several key innovations are introduced that allow the study of diverse models within an Isolation-with-Migration framework. The new method implements a 2-step analysis, with an initial Markov chain Monte Carlo (MCMC) phase that samples simple coalescent trees, followed by the calculation of the joint posterior density for the parameters of a demographic model. In step 1, the MCMC sampling phase, the method uses a reduced state space, consisting of coalescent trees without migration paths, and a simple importance sampling distribution without the demography of interest. Once obtained, a single sample of trees can be used in step 2 to calculate the joint posterior density for model parameters under multiple diverse demographic models, without having to repeat MCMC runs. Because migration paths are not included in the state space of the MCMC phase, but rather are handled by analytic integration in step 2 of the analysis, the method is scalable to a large number of loci with excellent MCMC mixing properties. With an implementation of the new method in the computer program MIST, we demonstrate the method's accuracy, scalability, and other advantages using simulated data and DNA sequences of two common chimpanzee subspecies: Pan troglodytes (P. t.) troglodytes and P. t. verus. © The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Hollow Microtube Resonators via Silicon Self-Assembly toward Subattogram Mass Sensing Applications.

PubMed

Kim, Joohyun; Song, Jungki; Kim, Kwangseok; Kim, Seokbeom; Song, Jihwan; Kim, Namsu; Khan, M Faheem; Zhang, Linan; Sader, John E; Park, Keunhan; Kim, Dongchoul; Thundat, Thomas; Lee, Jungchul

2016-03-09

Fluidic resonators with integrated microchannels (hollow resonators) are attractive for mass, density, and volume measurements of single micro/nanoparticles and cells, yet their widespread use is limited by the complexity of their fabrication. Here we report a simple and cost-effective approach for fabricating hollow microtube resonators. A prestructured silicon wafer is annealed at high temperature under a controlled atmosphere to form self-assembled buried cavities. The interiors of these cavities are oxidized to produce thin oxide tubes, following which the surrounding silicon material is selectively etched away to suspend the oxide tubes. This simple three-step process easily produces hollow microtube resonators. We report another innovation in the capping glass wafer where we integrate fluidic access channels and getter materials along with residual gas suction channels. Combined together, only five photolithographic steps and one bonding step are required to fabricate vacuum-packaged hollow microtube resonators that exhibit quality factors as high as ∼ 13,000. We take one step further to explore additionally attractive features including the ability to tune the device responsivity, changing the resonator material, and scaling down the resonator size. The resonator wall thickness of ∼ 120 nm and the channel hydraulic diameter of ∼ 60 nm are demonstrated solely by conventional microfabrication approaches. The unique characteristics of this new fabrication process facilitate the widespread use of hollow microtube resonators, their translation between diverse research fields, and the production of commercially viable devices.

Enzymatic production of single-molecule FISH and RNA capture probes.

PubMed

Gaspar, Imre; Wippich, Frank; Ephrussi, Anne

2017-10-01

Arrays of singly labeled short oligonucleotides that hybridize to a specific target revolutionized RNA biology, enabling quantitative, single-molecule microscopy analysis and high-efficiency RNA/RNP capture. Here, we describe a simple and efficient method that allows flexible functionalization of inexpensive DNA oligonucleotides by different fluorescent dyes or biotin using terminal deoxynucleotidyl transferase and custom-made functional group conjugated dideoxy-UTP. We show that (i) all steps of the oligonucleotide labeling-including conjugation, enzymatic synthesis, and product purification-can be performed in a standard biology laboratory, (ii) the process yields >90%, often >95% labeled product with minimal carryover of impurities, and (iii) the oligonucleotides can be labeled with different dyes or biotin, allowing single-molecule FISH, RNA affinity purification, and Northern blot analysis to be performed. © 2017 Gaspar et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

Validation of a single-stage fixed-rate step test for the prediction of maximal oxygen uptake in healthy adults.

PubMed

Hansen, Dominique; Jacobs, Nele; Thijs, Herbert; Dendale, Paul; Claes, Neree

2016-09-01

Healthcare professionals with limited access to ergospirometry remain in need of valid and simple submaximal exercise tests to predict maximal oxygen uptake (VO2max ). Despite previous validation studies concerning fixed-rate step tests, accurate equations for the estimation of VO2max remain to be formulated from a large sample of healthy adults between age 18-75 years (n > 100). The aim of this study was to develop a valid equation to estimate VO2max from a fixed-rate step test in a larger sample of healthy adults. A maximal ergospirometry test, with assessment of cardiopulmonary parameters and VO2max , and a 5-min fixed-rate single-stage step test were executed in 112 healthy adults (age 18-75 years). During the step test and subsequent recovery, heart rate was monitored continuously. By linear regression analysis, an equation to predict VO2max from the step test was formulated. This equation was assessed for level of agreement by displaying Bland-Altman plots and calculation of intraclass correlations with measured VO2max . Validity further was assessed by employing a Jackknife procedure. The linear regression analysis generated the following equation to predict VO2max (l min(-1) ) from the step test: 0·054(BMI)+0·612(gender)+3·359(body height in m)+0·019(fitness index)-0·012(HRmax)-0·011(age)-3·475. This equation explained 78% of the variance in measured VO2max (F = 66·15, P<0·001). The level of agreement and intraclass correlation was high (ICC = 0·94, P<0·001) between measured and predicted VO2max . From this study, a valid fixed-rate single-stage step test equation has been developed to estimate VO2max in healthy adults. This tool could be employed by healthcare professionals with limited access to ergospirometry. © 2015 Scandinavian Society of Clinical Physiology and Nuclear Medicine. Published by John Wiley & Sons Ltd.

An Improved Single-Step Cloning Strategy Simplifies the Agrobacterium tumefaciens-Mediated Transformation (ATMT)-Based Gene-Disruption Method for Verticillium dahliae.

PubMed

Wang, Sheng; Xing, Haiying; Hua, Chenlei; Guo, Hui-Shan; Zhang, Jie

2016-06-01

The soilborne fungal pathogen Verticillium dahliae infects a broad range of plant species to cause severe diseases. The availability of Verticillium genome sequences has provided opportunities for large-scale investigations of individual gene function in Verticillium strains using Agrobacterium tumefaciens-mediated transformation (ATMT)-based gene-disruption strategies. Traditional ATMT vectors require multiple cloning steps and elaborate characterization procedures to achieve successful gene replacement; thus, these vectors are not suitable for high-throughput ATMT-based gene deletion. Several advancements have been made that either involve simplification of the steps required for gene-deletion vector construction or increase the efficiency of the technique for rapid recombinant characterization. However, an ATMT binary vector that is both simple and efficient is still lacking. Here, we generated a USER-ATMT dual-selection (DS) binary vector, which combines both the advantages of the USER single-step cloning technique and the efficiency of the herpes simplex virus thymidine kinase negative-selection marker. Highly efficient deletion of three different genes in V. dahliae using the USER-ATMT-DS vector enabled verification that this newly-generated vector not only facilitates the cloning process but also simplifies the subsequent identification of fungal homologous recombinants. The results suggest that the USER-ATMT-DS vector is applicable for efficient gene deletion and suitable for large-scale gene deletion in V. dahliae.

Several steps/day indicators predict changes in anthropometric outcomes: HUB city steps

USDA-ARS?s Scientific Manuscript database

Walking for exercise remains the most frequently reported leisure-time activity, likely because it is simple, inexpensive, and easily incorporated into most people’s lifestyle. Pedometers are simple, convenient, and economical tools that can be used to quantify step-determined physical activity. F...

Measurement of intrahepatic pressure during radiofrequency ablation in porcine liver.

PubMed

Kawamoto, Chiaki; Yamauchi, Atsushi; Baba, Yoko; Kaneko, Keiko; Yakabi, Koji

2010-04-01

To identify the most effective procedures to avoid increased intrahepatic pressure during radiofrequency ablation, we evaluated different ablation methods. Laparotomy was performed in 19 pigs. Intrahepatic pressure was monitored using an invasive blood pressure monitor. Radiofrequency ablation was performed as follows: single-step standard ablation; single-step at 30 W; single-step at 70 W; 4-step at 30 W; 8-step at 30 W; 8-step at 70 W; and cooled-tip. The array was fully deployed in single-step methods. In the multi-step methods, the array was gradually deployed in four or eight steps. With the cooled-tip, ablation was performed by increasing output by 10 W/min, starting at 40 W. Intrahepatic pressure was as follows: single-step standard ablation, 154.5 +/- 30.9 mmHg; single-step at 30 W, 34.2 +/- 20.0 mmHg; single-step at 70 W, 46.7 +/- 24.3 mmHg; 4-step at 30 W, 42.3 +/- 17.9 mmHg; 8-step at 30 W, 24.1 +/- 18.2 mmHg; 8-step at 70 W, 47.5 +/- 31.5 mmHg; and cooled-tip, 114.5 +/- 16.6 mmHg. The radiofrequency ablation-induced area was spherical with single-step standard ablation, 4-step at 30 W, and 8-step at 30 W. Conversely, the ablated area was irregular with single-step at 30 W, single-step at 70 W, and 8-step at 70 W. The ablation time was significantly shorter for the multi-step method than for the single-step method. Increased intrahepatic pressure could be controlled using multi-step methods. From the shapes of the ablation area, 30-W 8-step expansions appear to be most suitable for radiofrequency ablation.

Modeling-independent elucidation of inactivation pathways in recombinant and native A-type Kv channels.

PubMed

Fineberg, Jeffrey D; Ritter, David M; Covarrubias, Manuel

2012-11-01

A-type voltage-gated K(+) (Kv) channels self-regulate their activity by inactivating directly from the open state (open-state inactivation [OSI]) or by inactivating before they open (closed-state inactivation [CSI]). To determine the inactivation pathways, it is often necessary to apply several pulse protocols, pore blockers, single-channel recording, and kinetic modeling. However, intrinsic hurdles may preclude the standardized application of these methods. Here, we implemented a simple method inspired by earlier studies of Na(+) channels to analyze macroscopic inactivation and conclusively deduce the pathways of inactivation of recombinant and native A-type Kv channels. We investigated two distinct A-type Kv channels expressed heterologously (Kv3.4 and Kv4.2 with accessory subunits) and their native counterparts in dorsal root ganglion and cerebellar granule neurons. This approach applies two conventional pulse protocols to examine inactivation induced by (a) a simple step (single-pulse inactivation) and (b) a conditioning step (double-pulse inactivation). Consistent with OSI, the rate of Kv3.4 inactivation (i.e., the negative first derivative of double-pulse inactivation) precisely superimposes on the profile of the Kv3.4 current evoked by a single pulse because the channels must open to inactivate. In contrast, the rate of Kv4.2 inactivation is asynchronous, already changing at earlier times relative to the profile of the Kv4.2 current evoked by a single pulse. Thus, Kv4.2 inactivation occurs uncoupled from channel opening, indicating CSI. Furthermore, the inactivation time constant versus voltage relation of Kv3.4 decreases monotonically with depolarization and levels off, whereas that of Kv4.2 exhibits a J-shape profile. We also manipulated the inactivation phenotype by changing the subunit composition and show how CSI and CSI combined with OSI might affect spiking properties in a full computational model of the hippocampal CA1 neuron. This work unambiguously elucidates contrasting inactivation pathways in neuronal A-type Kv channels and demonstrates how distinct pathways might impact neurophysiological activity.

Single molecule photobleaching (SMPB) technology for counting of RNA, DNA, protein and other molecules in nanoparticles and biological complexes by TIRF instrumentation.

PubMed

Zhang, Hui; Guo, Peixuan

2014-05-15

Direct counting of biomolecules within biological complexes or nanomachines is demanding. Single molecule counting using optical microscopy is challenging due to the diffraction limit. The single molecule photobleaching (SMPB) technology for direct counting developed by our team (Shu et al., 2007 [18]; Zhang et al., 2007 [19]) offers a simple and straightforward method to determine the stoichiometry of molecules or subunits within biocomplexes or nanomachines at nanometer scales. Stoichiometry is determined by real-time observation of the number of descending steps resulted from the photobleaching of individual fluorophore. This technology has now been used extensively for single molecule counting of protein, RNA, and other macromolecules in a variety of complexes or nanostructures. Here, we elucidate the SMPB technology, using the counting of RNA molecules within a bacteriophage phi29 DNA-packaging biomotor as an example. The method described here can be applied to the single molecule counting of other molecules in other systems. The construction of a concise, simple and economical single molecule total internal reflection fluorescence (TIRF) microscope combining prism-type and objective-type TIRF is described. The imaging system contains a deep-cooled sensitive EMCCD camera with single fluorophore detection sensitivity, a laser combiner for simultaneous dual-color excitation, and a Dual-View™ imager to split the multiple outcome signals to different detector channels based on their wavelengths. Methodology of the single molecule photobleaching assay used to elucidate the stoichiometry of RNA on phi29 DNA packaging motor and the mechanism of protein/RNA interaction are described. Different methods for single fluorophore labeling of RNA molecules are reviewed. The process of statistical modeling to reveal the true copy number of the biomolecules based on binomial distribution is also described. Copyright © 2014 Elsevier Inc. All rights reserved.

Quantum robots and environments

DOE Office of Scientific and Technical Information (OSTI.GOV)

Benioff, P.

1998-08-01

Quantum robots and their interactions with environments of quantum systems are described, and their study justified. A quantum robot is a mobile quantum system that includes an on-board quantum computer and needed ancillary systems. Quantum robots carry out tasks whose goals include specified changes in the state of the environment, or carrying out measurements on the environment. Each task is a sequence of alternating computation and action phases. Computation phase activites include determination of the action to be carried out in the next phase, and recording of information on neighborhood environmental system states. Action phase activities include motion of themore » quantum robot and changes in the neighborhood environment system states. Models of quantum robots and their interactions with environments are described using discrete space and time. A unitary step operator T that gives the single time step dynamics is associated with each task. T=T{sub a}+T{sub c} is a sum of action phase and computation phase step operators. Conditions that T{sub a} and T{sub c} should satisfy are given along with a description of the evolution as a sum over paths of completed phase input and output states. A simple example of a task{emdash}carrying out a measurement on a very simple environment{emdash}is analyzed in detail. A decision tree for the task is presented and discussed in terms of the sums over phase paths. It is seen that no definite times or durations are associated with the phase steps in the tree, and that the tree describes the successive phase steps in each path in the sum over phase paths. {copyright} {ital 1998} {ital The American Physical Society}« less

A simple approach to optimal control of invasive species.

PubMed

Hastings, Alan; Hall, Richard J; Taylor, Caz M

2006-12-01

The problem of invasive species and their control is one of the most pressing applied issues in ecology today. We developed simple approaches based on linear programming for determining the optimal removal strategies of different stage or age classes for control of invasive species that are still in a density-independent phase of growth. We illustrate the application of this method to the specific example of invasive Spartina alterniflora in Willapa Bay, WA. For all such systems, linear programming shows in general that the optimal strategy in any time step is to prioritize removal of a single age or stage class. The optimal strategy adjusts which class is the focus of control through time and can be much more cost effective than prioritizing removal of the same stage class each year.

Mirror gait retraining for the treatment of patellofemoral pain in female runners

PubMed Central

Willy, Richard W.; Scholz, John P.; Davis, Irene S.

2012-01-01

Background Abnormal hip mechanics are often implicated in female runners with patellofemoral pain. We sought to evaluate a simple gait retraining technique, using a full-length mirror, in female runners with patellofemoral pain and abnormal hip mechanics. Transfer of the new motor skill to the untrained tasks of single leg squat and step descent was also evaluated. Methods Ten female runners with patellofemoral pain completed 8 sessions of mirror and verbal feedback on their lower extremity alignment during treadmill running. During the last 4 sessions, mirror and verbal feedback were progressively removed. Hip mechanics were assessed during running gait, a single leg squat and a step descent, both pre- and post-retraining. Subjects returned to their normal running routines and analyses were repeated at 1-month and 3-month post-retraining. Data were analyzed via repeated measures analysis of variance. Findings Subjects reduced peaks of hip adduction, contralateral pelvic drop, and hip abduction moment during running (P<0.05, effect size=0.69–2.91). Skill transfer to single leg squatting and step descent was noted (P<0.05, effect size=0.91–1.35). At 1 and 3 months post retraining, most mechanics were maintained in the absence of continued feedback. Subjects reported improvements in pain and function (P<0.05, effect size=3.81–7.61) and maintained through 3 months post retraining. Interpretation Mirror gait retraining was effective in improving mechanics and measures of pain and function. Skill transfer to the untrained tasks of squatting and step descent indicated that a higher level of motor learning had occurred. Extended follow-up is needed to determine the long term efficacy of this treatment. PMID:22917625

Human Mitochondrial RNA Polymerase: Evaluation of the Single-Nucleotide-Addition Cycle on Synthetic RNA/DNA Scaffolds

PubMed Central

Smidansky, Eric D.; Arnold, Jamie J.; Reynolds, Shelley L.; Cameron, Craig E.

2013-01-01

The human mitochondrial RNA polymerase (h-mtRNAP) serves as both the transcriptase for expression and the primase for replication of mitochondrial DNA. As such, the enzyme is of fundamental importance to cellular energy metabolism, and defects in its function may be related to human disease states. Here we describe in vitro analysis of the h-mtRNAP kinetic mechanism for single, correct nucleotide incorporation. This was made possible by the development of efficient methods for expression and purification of h-mtRNAP using a bacterial system and by utilization of assays that rely on simple, synthetic RNA/DNA scaffolds without the need for mitochondrial transcription accessory proteins. We find that h-mtRNAP accomplishes single-nucleotide incorporation by using the same core steps, including conformational change steps before and after chemistry, that are prototypical for most types of nucleic acid polymerases. The polymerase binds to scaffolds via a two-step mechanism consisting of a fast initial-encounter step followed by a much slower isomerization that leads to catalytic competence. A substantial solvent deuterium kinetic isotope effect was observed for the forward reaction, but none was detectable for the reverse reaction, suggesting that chemistry is at least partially rate-limiting in the forward direction but not in the reverse. h-mtRNAP appears to exercise much more stringent surveillance over base than over sugar in determining the correctness of a nucleotide. The utility of developing the robust in vitro assays described here and of establishing a baseline of kinetic performance for the wild-type enzyme is that biological questions concerning h-mtRNAP may now begin to be addressed. PMID:21548588

A comparison of simple global kinetic models for coal devolatilization with the CPD model

DOE PAGES

Richards, Andrew P.; Fletcher, Thomas H.

2016-08-01

Simulations of coal combustors and gasifiers generally cannot incorporate the complexities of advanced pyrolysis models, and hence there is interest in evaluating simpler models over ranges of temperature and heating rate that are applicable to the furnace of interest. In this paper, six different simple model forms are compared to predictions made by the Chemical Percolation Devolatilization (CPD) model. The model forms included three modified one-step models, a simple two-step model, and two new modified two-step models. These simple model forms were compared over a wide range of heating rates (5 × 10 3 to 10 6 K/s) at finalmore » temperatures up to 1600 K. Comparisons were made of total volatiles yield as a function of temperature, as well as the ultimate volatiles yield. Advantages and disadvantages for each simple model form are discussed. In conclusion, a modified two-step model with distributed activation energies seems to give the best agreement with CPD model predictions (with the fewest tunable parameters).« less

Regularized lattice Boltzmann model for immiscible two-phase flows with power-law rheology

NASA Astrophysics Data System (ADS)

Ba, Yan; Wang, Ningning; Liu, Haihu; Li, Qiang; He, Guoqiang

2018-03-01

In this work, a regularized lattice Boltzmann color-gradient model is developed for the simulation of immiscible two-phase flows with power-law rheology. This model is as simple as the Bhatnagar-Gross-Krook (BGK) color-gradient model except that an additional regularization step is introduced prior to the collision step. In the regularization step, the pseudo-inverse method is adopted as an alternative solution for the nonequilibrium part of the total distribution function, and it can be easily extended to other discrete velocity models no matter whether a forcing term is considered or not. The obtained expressions for the nonequilibrium part are merely related to macroscopic variables and velocity gradients that can be evaluated locally. Several numerical examples, including the single-phase and two-phase layered power-law fluid flows between two parallel plates, and the droplet deformation and breakup in a simple shear flow, are conducted to test the capability and accuracy of the proposed color-gradient model. Results show that the present model is more stable and accurate than the BGK color-gradient model for power-law fluids with a wide range of power-law indices. Compared to its multiple-relaxation-time counterpart, the present model can increase the computing efficiency by around 15%, while keeping the same accuracy and stability. Also, the present model is found to be capable of reasonably predicting the critical capillary number of droplet breakup.

Compact mass spectrometer for plasma discharge ion analysis

DOEpatents

Tuszewski, M.G.

1997-07-22

A mass spectrometer and methods are disclosed for mass spectrometry which are useful in characterizing a plasma. This mass spectrometer for determining type and quantity of ions present in a plasma is simple, compact, and inexpensive. It accomplishes mass analysis in a single step, rather than the usual two-step process comprised of ion extraction followed by mass filtering. Ions are captured by a measuring element placed in a plasma and accelerated by a known applied voltage. Captured ions are bent into near-circular orbits by a magnetic field such that they strike a collector, producing an electric current. Ion orbits vary with applied voltage and proton mass ratio of the ions, so that ion species may be identified. Current flow provides an indication of quantity of ions striking the collector. 7 figs.

Compact mass spectrometer for plasma discharge ion analysis

DOEpatents

Tuszewski, Michel G.

1997-01-01

A mass spectrometer and methods for mass spectrometry which are useful in characterizing a plasma. This mass spectrometer for determining type and quantity of ions present in a plasma is simple, compact, and inexpensive. It accomplishes mass analysis in a single step, rather than the usual two-step process comprised of ion extraction followed by mass filtering. Ions are captured by a measuring element placed in a plasma and accelerated by a known applied voltage. Captured ions are bent into near-circular orbits by a magnetic field such that they strike a collector, producing an electric current. Ion orbits vary with applied voltage and proton mass ratio of the ions, so that ion species may be identified. Current flow provides an indication of quantity of ions striking the collector.

Simplified Large-Scale Refolding, Purification, and Characterization of Recombinant Human Granulocyte-Colony Stimulating Factor in Escherichia coli

PubMed Central

Kim, Chang Kyu; Lee, Chi Ho; Lee, Seung-Bae; Oh, Jae-Wook

2013-01-01

Granulocyte-colony stimulating factor (G-CSF) is a pleiotropic cytokine that stimulates the development of committed hematopoietic progenitor cells and enhances the functional activity of mature cells. Here, we report a simplified method for fed-batch culture as well as the purification of recombinant human (rh) G-CSF. The new system for rhG-CSF purification was performed using not only temperature shift strategy without isopropyl-l-thio-β-d-galactoside (IPTG) induction but also the purification method by a single step of prep-HPLC after the pH precipitation of the refolded samples. Through these processes, the final cell density and overall yield of homogenous rhG-CSF were obtained 42.8 g as dry cell weights, 1.75 g as purified active proteins, from 1 L culture broth, respectively. The purity of rhG-CSF was finally 99% since the isoforms of rhG-CSF could be separated through the prep-HPLC step. The result of biological activity indicated that purified rhG-CSF has a similar profile to the World Health Organization (WHO) 2nd International Standard for G-CSF. Taken together, our results demonstrate that the simple purification through a single step of prep-HPLC may be valuable for the industrial-scale production of biologically active proteins. PMID:24224041

Understanding the allocation of attention when faced with varying perceptual load in partial report: a computational approach.

PubMed

Kyllingsbæk, Søren; Sy, Jocelyn L; Giesbrecht, Barry

2011-05-01

The allocation of visual processing capacity is a key topic in studies and theories of visual attention. The load theory of Lavie (1995) proposes that allocation happens in two steps where processing resources are first allocated to task-relevant stimuli and secondly remaining capacity 'spills over' to task-irrelevant distractors. In contrast, the Theory of Visual Attention (TVA) proposed by Bundesen (1990) assumes that allocation happens in a single step where processing capacity is allocated to all stimuli, both task-relevant and task-irrelevant, in proportion to their relative attentional weight. Here we present data from two partial report experiments where we varied the number and discriminability of the task-irrelevant stimuli (Experiment 1) and perceptual load (Experiment 2). The TVA fitted the data of the two experiments well thus favoring the simple explanation with a single step of capacity allocation. We also show that the effects of varying perceptual load can only be explained by a combined effect of allocation of processing capacity as well as limits in visual working memory. Finally, we link the results to processing capacity understood at the neural level based on the neural theory of visual attention by Bundesen et al. (2005). Copyright © 2010 Elsevier Ltd. All rights reserved.

Computer-Aided Design of RNA Origami Structures.

PubMed

Sparvath, Steffen L; Geary, Cody W; Andersen, Ebbe S

2017-01-01

RNA nanostructures can be used as scaffolds to organize, combine, and control molecular functionalities, with great potential for applications in nanomedicine and synthetic biology. The single-stranded RNA origami method allows RNA nanostructures to be folded as they are transcribed by the RNA polymerase. RNA origami structures provide a stable framework that can be decorated with functional RNA elements such as riboswitches, ribozymes, interaction sites, and aptamers for binding small molecules or protein targets. The rich library of RNA structural and functional elements combined with the possibility to attach proteins through aptamer-based binding creates virtually limitless possibilities for constructing advanced RNA-based nanodevices.In this chapter we provide a detailed protocol for the single-stranded RNA origami design method using a simple 2-helix tall structure as an example. The first step involves 3D modeling of a double-crossover between two RNA double helices, followed by decoration with tertiary motifs. The second step deals with the construction of a 2D blueprint describing the secondary structure and sequence constraints that serves as the input for computer programs. In the third step, computer programs are used to design RNA sequences that are compatible with the structure, and the resulting outputs are evaluated and converted into DNA sequences to order.

Elastic-net regularization approaches for genome-wide association studies of rheumatoid arthritis.

PubMed

Cho, Seoae; Kim, Haseong; Oh, Sohee; Kim, Kyunga; Park, Taesung

2009-12-15

The current trend in genome-wide association studies is to identify regions where the true disease-causing genes may lie by evaluating thousands of single-nucleotide polymorphisms (SNPs) across the whole genome. However, many challenges exist in detecting disease-causing genes among the thousands of SNPs. Examples include multicollinearity and multiple testing issues, especially when a large number of correlated SNPs are simultaneously tested. Multicollinearity can often occur when predictor variables in a multiple regression model are highly correlated, and can cause imprecise estimation of association. In this study, we propose a simple stepwise procedure that identifies disease-causing SNPs simultaneously by employing elastic-net regularization, a variable selection method that allows one to address multicollinearity. At Step 1, the single-marker association analysis was conducted to screen SNPs. At Step 2, the multiple-marker association was scanned based on the elastic-net regularization. The proposed approach was applied to the rheumatoid arthritis (RA) case-control data set of Genetic Analysis Workshop 16. While the selected SNPs at the screening step are located mostly on chromosome 6, the elastic-net approach identified putative RA-related SNPs on other chromosomes in an increased proportion. For some of those putative RA-related SNPs, we identified the interactions with sex, a well known factor affecting RA susceptibility.

Submicron diameter single crystal sapphire optical fiber

DOE PAGES

Hill, Cary; Homa, Daniel; Liu, Bo; ...

2014-10-02

In this work, a submicron-diameter single crystal sapphire optical fiber was demonstrated via wet acid etching at elevated temperatures. Etch rates on the order 2.3 µm/hr were achievable with a 3:1 molar ratio sulfuric-phosphoric acid solution maintained at a temperature of 343°C. A sapphire fiber with an approximate diameter of 800 nm was successfully fabricated from a commercially available fiber with an original diameter of 50 µm. The simple and controllable etching technique provides a feasible approach to the fabrication of unique waveguide structures via traditional silica masking techniques. The ability to tailor the geometry of sapphire optical fibers ismore » the first step in achieving optical and sensing performance on par with its fused silica counterpart.« less

Discrimination of Geographical Origin of Asian Garlic Using Isotopic and Chemical Datasets under Stepwise Principal Component Analysis.

PubMed

Liu, Tsang-Sen; Lin, Jhen-Nan; Peng, Tsung-Ren

2018-01-16

Isotopic compositions of δ 2 H, δ 18 O, δ 13 C, and δ 15 N and concentrations of 22 trace elements from garlic samples were analyzed and processed with stepwise principal component analysis (PCA) to discriminate garlic's country of origin among Asian regions including South Korea, Vietnam, Taiwan, and China. Results indicate that there is no single trace-element concentration or isotopic composition that can accomplish the study's purpose and the stepwise PCA approach proposed does allow for discrimination between countries on a regional basis. Sequentially, Step-1 PCA distinguishes garlic's country of origin among Taiwanese, South Korean, and Vietnamese samples; Step-2 PCA discriminates Chinese garlic from South Korean garlic; and Step-3 and Step-4 PCA, Chinese garlic from Vietnamese garlic. In model tests, countries of origin of all audit samples were correctly discriminated by stepwise PCA. Consequently, this study demonstrates that stepwise PCA as applied is a simple and effective approach to discriminating country of origin among Asian garlics. © 2018 American Academy of Forensic Sciences.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bowman, Adam J.; Scherrer, Joseph R.; Reiserer, Ronald S., E-mail: ron.reiserer@vanderbilt.edu

We present a simple apparatus for improved surface modification of polydimethylsiloxane (PDMS) microfluidic devices. A single treatment chamber for plasma activation and chemical/physical vapor deposition steps minimizes the time-dependent degradation of surface activation that is inherent in multi-chamber techniques. Contamination and deposition irregularities are also minimized by conducting plasma activation and treatment phases in the same vacuum environment. An inductively coupled plasma driver allows for interchangeable treatment chambers. Atomic force microscopy confirms that silane deposition on PDMS gives much better surface quality than standard deposition methods, which yield a higher local roughness and pronounced irregularities in the surface.

A step-by-step solution for embedding user-controlled cines into educational Web pages.

PubMed

Cornfeld, Daniel

2008-03-01

The objective of this article is to introduce a simple method for embedding user-controlled cines into a Web page using a simple JavaScript. Step-by-step instructions are included and the source code is made available. This technique allows the creation of portable Web pages that allow the user to scroll through cases as if seated at a PACS workstation. A simple JavaScript allows scrollable image stacks to be included on Web pages. With this technique, you can quickly and easily incorporate entire stacks of CT or MR images into online teaching files. This technique has the potential for use in case presentations, online didactics, teaching archives, and resident testing.

Preparative isolation, fast centrifugal partition chromatography purification and biological activity of cajaflavanone from Derris ferruginea stems.

PubMed

Morel, Sylvie; Landreau, Anne; Nguyen, Van Hung; Derbré, Séverine; Grellier, Philippe; Pape, Patrice Le; Pagniez, Fabrice; Litaudon, Marc; Richomme, Pascal

2012-01-01

The Derris genus is known to contain flavonoid derivatives, including prenylated flavanones and isoflavonoids such as rotenoids, which are generally associated with significant biological activity. To develop an efficient preparative isolation procedure for bioactive cajaflavanone. Fast centrifugal partition chromatography (FCPC) was optimised to purify cajaflavanone from Derris ferruginea stems in a single step as compared to fractionation from the cyclohexane extract by successive conventional solid-liquid chromatography procedures. The purification yield, purity, time and solvent consumption per procedure are described. The anti-fungal, anti-bacterial, anti-leishmanial, anti-plasmodial, anti-oxidant activities and the inhibition of advanced glycation end-products (AGEs) by cajaflavanone accumulation are described. FCPC enabled cajaflavanone purification in a single separation step, yielding sufficient quantities to perform in vitro biological screening. Interestingly, cajaflavanone had an inhibitory effect on the formation of AGEs, without displaying any in vitro anti-oxidant activity. A simple and efficient procedure, in comparison with other preparative methods, for bioactive cajaflavone purification has been developed using FCPC. Copyright © 2011 John Wiley & Sons, Ltd.

Processive movement of single kinesins on crowded microtubules visualized using quantum dots

PubMed Central

Seitz, Arne; Surrey, Thomas

2006-01-01

Kinesin-1 is a processive molecular motor transporting cargo along microtubules. Inside cells, several motors and microtubule-associated proteins compete for binding to microtubules. Therefore, the question arises how processive movement of kinesin-1 is affected by crowding on the microtubule. Here we use total internal reflection fluorescence microscopy to image in vitro the runs of single quantum dot-labelled kinesins on crowded microtubules under steady-state conditions and to measure the degree of crowding on a microtubule at steady-state. We find that the runs of kinesins are little affected by high kinesin densities on a microtubule. However, the presence of high densities of a mutant kinesin that is not able to step efficiently reduces the average speed of wild-type kinesin, while hardly changing its processivity. This indicates that kinesin waits in a strongly bound state on the microtubule when encountering an obstacle until the obstacle unbinds and frees the binding site for kinesin's next step. A simple kinetic model can explain quantitatively the behaviour of kinesin under both crowding conditions. PMID:16407972

Laser-induced forward transfer for flip-chip packaging of single dies.

PubMed

Kaur, Kamal S; Van Steenberge, Geert

2015-03-20

Flip-chip (FC) packaging is a key technology for realizing high performance, ultra-miniaturized and high-density circuits in the micro-electronics industry. In this technique the chip and/or the substrate is bumped and the two are bonded via these conductive bumps. Many bumping techniques have been developed and intensively investigated since the introduction of the FC technology in 1960(1) such as stencil printing, stud bumping, evaporation and electroless/electroplating2. Despite the progress that these methods have made they all suffer from one or more than one drawbacks that need to be addressed such as cost, complex processing steps, high processing temperatures, manufacturing time and most importantly the lack of flexibility. In this paper, we demonstrate a simple and cost-effective laser-based bump forming technique known as Laser-induced Forward Transfer (LIFT)3. Using the LIFT technique a wide range of bump materials can be printed in a single-step with great flexibility, high speed and accuracy at RT. In addition, LIFT enables the bumping and bonding down to chip-scale, which is critical for fabricating ultra-miniature circuitry.

Determination of pyrophosphate and sulfate using polyhexamethylene guanidine hydrochloride-stabilized silver nanoparticles.

PubMed

Terenteva, E A; Apyari, V V; Dmitrienko, S G; Garshev, A V; Volkov, P A; Zolotov, Yu A

2018-04-01

Positively charged polyhexamethylene guanidine hydrochloride-stabilized silver nanoparticles (PHMG-AgNPs) were prepared and applied as a colorimetric probe for single-step determination of pyrophosphate and sulfate. The approach is based on the nanoparticles aggregation leading to change in their absorption spectra and color of the solution. Due to both electrostatic and steric stabilization these nanoparticles show decreased sensitivity relatively to many common anions, which allows for simple and rapid direct single-step determination of pyrophosphate and sulfate. Effects of different factors (time of interaction, pH, concentrations of anions and the nanoparticles) on aggregation of PHMG-AgNPs and analytical performance of the procedure were investigated. The method allows for the determination of pyrophosphate and sulfate in the range of 0.16-2μgmL -1 and 20-80μgmL -1 with RSD of 2-5%, respectively. The analysis can be performed using either spectrophotometry or naked-eye detection. Practical application of the method was shown by the example of pyrophosphate determination in baking powder sample. Copyright © 2017 Elsevier B.V. All rights reserved.

Single-step collision-free trajectory planning of biped climbing robots in spatial trusses.

PubMed

Zhu, Haifei; Guan, Yisheng; Chen, Shengjun; Su, Manjia; Zhang, Hong

For a biped climbing robot with dual grippers to climb poles, trusses or trees, feasible collision-free climbing motion is inevitable and essential. In this paper, we utilize the sampling-based algorithm, Bi-RRT, to plan single-step collision-free motion for biped climbing robots in spatial trusses. To deal with the orientation limit of a 5-DoF biped climbing robot, a new state representation along with corresponding operations including sampling, metric calculation and interpolation is presented. A simple but effective model of a biped climbing robot in trusses is proposed, through which the motion planning of one climbing cycle is transformed to that of a manipulator. In addition, the pre- and post-processes are introduced to expedite the convergence of the Bi-RRT algorithm and to ensure the safe motion of the climbing robot near poles as well. The piecewise linear paths are smoothed by utilizing cubic B-spline curve fitting. The effectiveness and efficiency of the presented Bi-RRT algorithm for climbing motion planning are verified by simulations.

Dye sensitized solar cells using catalytically active CuO-ZnO nanocomposite synthesized by single step method

NASA Astrophysics Data System (ADS)

Abraham, Nelsa; Rufus, Alex; Unni, C.; Philip, Daizy

2018-07-01

In the present paper we report a low cost, single step preparation method for the synthesis of CuO-ZnO nanocomposite through simple co-precipitation technique using oxalic acid. To have a better idea about the deviations brought about by the inclusion of CuO in ZnO lattice, pure ZnO nanoparticles synthesized from 0.1 M solutions were also investigated. X-ray diffraction studies showed that the composite contains only hexagonal wurtzite ZnO and monoclinic CuO structures. The magnetic studies of CuO-ZnO heterostructures were also conducted in order to elucidate the source of the ferromagnetism observed at room temperature. The catalytic efficiency of the as prepared nanocomposite was estimated by the degradation of methylene blue and eosin yellowish which also shows its suitability as a promising candidate in waste water treatment. The effect of chenodeoxycholic acid as a co-adsorbent in the performance of dye sensitized solar cells fabricated using the synthesized ZnO and the nanocomposite was also studied and significant improvement in photovoltaic performance has been obtained for nanocomposite based solar cell.

Very large scale monoclonal antibody purification: the case for conventional unit operations.

PubMed

Kelley, Brian

2007-01-01

Technology development initiatives targeted for monoclonal antibody purification may be motivated by manufacturing limitations and are often aimed at solving current and future process bottlenecks. A subject under debate in many biotechnology companies is whether conventional unit operations such as chromatography will eventually become limiting for the production of recombinant protein therapeutics. An evaluation of the potential limitations of process chromatography and filtration using today's commercially available resins and membranes was conducted for a conceptual process scaled to produce 10 tons of monoclonal antibody per year from a single manufacturing plant, a scale representing one of the world's largest single-plant capacities for cGMP protein production. The process employs a simple, efficient purification train using only two chromatographic and two ultrafiltration steps, modeled after a platform antibody purification train that has generated 10 kg batches in clinical production. Based on analyses of cost of goods and the production capacity of this very large scale purification process, it is unlikely that non-conventional downstream unit operations would be needed to replace conventional chromatographic and filtration separation steps, at least for recombinant antibodies.

On the Possibility of Ill-Conditioned Covariance Matrices in the First-Order Two-Step Estimator

NASA Technical Reports Server (NTRS)

Garrison, James L.; Axelrod, Penina; Kasdin, N. Jeremy

1997-01-01

The first-order two-step nonlinear estimator, when applied to a problem of orbital navigation, is found to occasionally produce first step covariance matrices with very low eigenvalues at certain trajectory points. This anomaly is the result of the linear approximation to the first step covariance propagation. The study of this anomaly begins with expressing the propagation of the first and second step covariance matrices in terms of a single matrix. This matrix is shown to have a rank equal to the difference between the number of first step states and the number of second step states. Furthermore, under some simplifying assumptions, it is found that the basis of the column space of this matrix remains fixed once the filter has removed the large initial state error. A test matrix containing the basis of this column space and the partial derivative matrix relating first and second step states is derived. This square test matrix, which has dimensions equal to the number of first step states, numerically drops rank at the same locations that the first step covariance does. It is formulated in terms of a set of constant vectors (the basis) and a matrix which can be computed from a reference trajectory (the partial derivative matrix). A simple example problem involving dynamics which are described by two states and a range measurement illustrate the cause of this anomaly and the application of the aforementioned numerical test in more detail.

Effect of the Four-Step Learning Cycle Model on Students' Understanding of Concepts Related to Simple Harmonic Motion

ERIC Educational Resources Information Center

Madu, B. C.

2012-01-01

The study explored the efficacy of four-step (4-E) learning cycle approach on students understanding of concepts related to Simple Harmonic Motion (SHM). 124 students (63 for experimental group and 61 for control group) participated in the study. The students' views and ideas in simple Harmonic Achievement test were analyzed qualitatively. The…

Reliable and cost effective design of intermetallic Ni2Si nanowires and direct characterization of its mechanical properties

NASA Astrophysics Data System (ADS)

Han, Seung Zeon; Kang, Joonhee; Kim, Sung-Dae; Choi, Si-Young; Kim, Hyung Giun; Lee, Jehyun; Kim, Kwangho; Lim, Sung Hwan; Han, Byungchan

2015-10-01

We report that a single crystal Ni2Si nanowire (NW) of intermetallic compound can be reliably designed using simple three-step processes: casting a ternary Cu-Ni-Si alloy, nucleate and growth of Ni2Si NWs as embedded in the alloy matrix via designing discontinuous precipitation (DP) of Ni2Si nanoparticles and thermal aging, and finally chemical etching to decouple the Ni2Si NWs from the alloy matrix. By direct application of uniaxial tensile tests to the Ni2Si NW we characterize its mechanical properties, which were rarely reported in previous literatures. Using integrated studies of first principles density functional theory (DFT) calculations, high-resolution transmission electron microscopy (HRTEM), and energy-dispersive X-ray spectroscopy (EDX) we accurately validate the experimental measurements. Our results indicate that our simple three-step method enables to design brittle Ni2Si NW with high tensile strength of 3.0 GPa and elastic modulus of 60.6 GPa. We propose that the systematic methodology pursued in this paper significantly contributes to opening innovative processes to design various kinds of low dimensional nanomaterials leading to advancement of frontiers in nanotechnology and related industry sectors.

Reliable and cost effective design of intermetallic Ni2Si nanowires and direct characterization of its mechanical properties.

PubMed

Han, Seung Zeon; Kang, Joonhee; Kim, Sung-Dae; Choi, Si-Young; Kim, Hyung Giun; Lee, Jehyun; Kim, Kwangho; Lim, Sung Hwan; Han, Byungchan

2015-10-12

We report that a single crystal Ni2Si nanowire (NW) of intermetallic compound can be reliably designed using simple three-step processes: casting a ternary Cu-Ni-Si alloy, nucleate and growth of Ni2Si NWs as embedded in the alloy matrix via designing discontinuous precipitation (DP) of Ni2Si nanoparticles and thermal aging, and finally chemical etching to decouple the Ni2Si NWs from the alloy matrix. By direct application of uniaxial tensile tests to the Ni2Si NW we characterize its mechanical properties, which were rarely reported in previous literatures. Using integrated studies of first principles density functional theory (DFT) calculations, high-resolution transmission electron microscopy (HRTEM), and energy-dispersive X-ray spectroscopy (EDX) we accurately validate the experimental measurements. Our results indicate that our simple three-step method enables to design brittle Ni2Si NW with high tensile strength of 3.0 GPa and elastic modulus of 60.6 GPa. We propose that the systematic methodology pursued in this paper significantly contributes to opening innovative processes to design various kinds of low dimensional nanomaterials leading to advancement of frontiers in nanotechnology and related industry sectors.

A numerical method for computing unsteady 2-D boundary layer flows

NASA Technical Reports Server (NTRS)

Krainer, Andreas

1988-01-01

A numerical method for computing unsteady two-dimensional boundary layers in incompressible laminar and turbulent flows is described and applied to a single airfoil changing its incidence angle in time. The solution procedure adopts a first order panel method with a simple wake model to solve for the inviscid part of the flow, and an implicit finite difference method for the viscous part of the flow. Both procedures integrate in time in a step-by-step fashion, in the course of which each step involves the solution of the elliptic Laplace equation and the solution of the parabolic boundary layer equations. The Reynolds shear stress term of the boundary layer equations is modeled by an algebraic eddy viscosity closure. The location of transition is predicted by an empirical data correlation originating from Michel. Since transition and turbulence modeling are key factors in the prediction of viscous flows, their accuracy will be of dominant influence to the overall results.

Motofit - integrating neutron reflectometry acquisition, reduction and analysis into one, easy to use, package

NASA Astrophysics Data System (ADS)

Nelson, Andrew

2010-11-01

The efficient use of complex neutron scattering instruments is often hindered by the complex nature of their operating software. This complexity exists at each experimental step: data acquisition, reduction and analysis, with each step being as important as the previous. For example, whilst command line interfaces are powerful at automated acquisition they often reduce accessibility by novice users and sometimes reduce the efficiency for advanced users. One solution to this is the development of a graphical user interface which allows the user to operate the instrument by a simple and intuitive "push button" approach. This approach was taken by the Motofit software package for analysis of multiple contrast reflectometry data. Here we describe the extension of this package to cover the data acquisition and reduction steps for the Platypus time-of-flight neutron reflectometer. Consequently, the complete operation of an instrument is integrated into a single, easy to use, program, leading to efficient instrument usage.

Direct current injection and thermocapillary flow for purification of aligned arrays of single-walled carbon nanotubes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Xie, Xu; Islam, Ahmad E.; Seabron, Eric

2015-04-07

Aligned arrays of semiconducting single-walled carbon nanotubes (s-SWNTs) represent ideal configurations for use of this class of material in high performance electronics. Development of means for removing the metallic SWNTs (m-SWNTs) in as-grown arrays represents an essential challenge. Here, we introduce a simple scheme that achieves this type of purification using direct, selective current injection through interdigitated electrodes into the m-SWNTs, to allow their complete removal using processes of thermocapillarity and dry etching. Experiments and numerical simulations establish the fundamental aspects that lead to selectivity in this process, thereby setting design rules for optimization. Single-step purification of arrays that includemore » thousands of SWNTs demonstrates the effectiveness and simplicity of the procedures. The result is a practical route to large-area aligned arrays of purely s-SWNTs with low-cost experimental setups.« less

Emergence of an optimal search strategy from a simple random walk

PubMed Central

Sakiyama, Tomoko; Gunji, Yukio-Pegio

2013-01-01

In reports addressing animal foraging strategies, it has been stated that Lévy-like algorithms represent an optimal search strategy in an unknown environment, because of their super-diffusion properties and power-law-distributed step lengths. Here, starting with a simple random walk algorithm, which offers the agent a randomly determined direction at each time step with a fixed move length, we investigated how flexible exploration is achieved if an agent alters its randomly determined next step forward and the rule that controls its random movement based on its own directional moving experiences. We showed that our algorithm led to an effective food-searching performance compared with a simple random walk algorithm and exhibited super-diffusion properties, despite the uniform step lengths. Moreover, our algorithm exhibited a power-law distribution independent of uniform step lengths. PMID:23804445

Emergence of an optimal search strategy from a simple random walk.

PubMed

Sakiyama, Tomoko; Gunji, Yukio-Pegio

2013-09-06

In reports addressing animal foraging strategies, it has been stated that Lévy-like algorithms represent an optimal search strategy in an unknown environment, because of their super-diffusion properties and power-law-distributed step lengths. Here, starting with a simple random walk algorithm, which offers the agent a randomly determined direction at each time step with a fixed move length, we investigated how flexible exploration is achieved if an agent alters its randomly determined next step forward and the rule that controls its random movement based on its own directional moving experiences. We showed that our algorithm led to an effective food-searching performance compared with a simple random walk algorithm and exhibited super-diffusion properties, despite the uniform step lengths. Moreover, our algorithm exhibited a power-law distribution independent of uniform step lengths.

Considering dominance in reduced single-step genomic evaluations.

PubMed

Ertl, J; Edel, C; Pimentel, E C G; Emmerling, R; Götz, K-U

2018-06-01

Single-step models including dominance can be an enormous computational task and can even be prohibitive for practical application. In this study, we try to answer the question whether a reduced single-step model is able to estimate breeding values of bulls and breeding values, dominance deviations and total genetic values of cows with acceptable quality. Genetic values and phenotypes were simulated (500 repetitions) for a small Fleckvieh pedigree consisting of 371 bulls (180 thereof genotyped) and 553 cows (40 thereof genotyped). This pedigree was virtually extended for 2,407 non-genotyped daughters. Genetic values were estimated with the single-step model and with different reduced single-step models. Including more relatives of genotyped cows in the reduced single-step model resulted in a better agreement of results with the single-step model. Accuracies of genetic values were largest with single-step and smallest with reduced single-step when only the cows genotyped were modelled. The results indicate that a reduced single-step model is suitable to estimate breeding values of bulls and breeding values, dominance deviations and total genetic values of cows with acceptable quality. © 2018 Blackwell Verlag GmbH.

Zero-bias microwave detectors based on array of nanorectifiers coupled with a dipole antenna

NASA Astrophysics Data System (ADS)

Kasjoo, Shahrir R.; Singh, Arun K.; Mat Isa, Siti S.; Ramli, Muhammad M.; Mohamad Isa, Muammar; Ahmad, Norhawati; Mohd Nor, Nurul I.; Khalid, Nazuhusna; Song, Ai Min

2016-04-01

We report on zero-bias microwave detection using a large array of unipolar nanodevices, known as the self-switching diodes (SSDs). The large array was realized in a single lithography step without the need of interconnection layers, hence allowing for a simple and low-cost fabrication process. The SSD array was coupled with a narrowband dipole antenna with a resonant frequency of 890 MHz, to form a simple rectenna (rectifying antenna). The extrinsic voltage responsivity and noise-equivalent-power (NEP) of the rectenna were ∼70 V/W and ∼0.18 nW/Hz1/2, respectively, measured in the far-field region at unbiased condition. Nevertheless, the estimated intrinsic voltage responsivity can achieve up to ∼5 kV/W with NEP of ∼2.6 pW/Hz1/2.

A single-step simultaneous protein staining procedure for polyacrylamide gels and nitrocellulose membranes by Alta during western blot analysis.

PubMed

Pal, Jayanta K; Berwal, Sunil K; Soni, Rupali N

2012-01-01

A simple method for staining of proteins simultaneously on sodium dodecyl sulfate (SDS) polyacrylamide gels and nitrocellulose membranes by Alta during western blot analysis is described. A 5% solution of Alta, a commercially available cosmetic preparation, is added in the upper tank buffer during electrophoresis. On completion of electrophoresis, the gel is washed in distilled water and viewed on a white light plate and a transilluminator to photograph the protein profiles. The gel is processed for western blot transfer of proteins onto a nitrocellulose membrane, and upon completion, the protein profiles on the membrane are viewed and photographed as stated above. The membrane can then be processed for immunostaining as per the standard procedure. Thus, the staining procedure using Alta is simple, rapid (without any need of destaining), and cost-effective.

Analysis and design of a standardized control module for switching regulators

NASA Astrophysics Data System (ADS)

Lee, F. C.; Mahmoud, M. F.; Yu, Y.; Kolecki, J. C.

1982-07-01

Three basic switching regulators: buck, boost, and buck/boost, employing a multiloop standardized control module (SCM) were characterized by a common small signal block diagram. Employing the unified model, regulator performances such as stability, audiosusceptibility, output impedance, and step load transient are analyzed and key performance indexes are expressed in simple analytical forms. More importantly, the performance characteristics of all three regulators are shown to enjoy common properties due to the unique SCM control scheme which nullifies the positive zero and provides adaptive compensation to the moving poles of the boost and buck/boost converters. This allows a simple unified design procedure to be devised for selecting the key SCM control parameters for an arbitrarily given power stage configuration and parameter values, such that all regulator performance specifications can be met and optimized concurrently in a single design attempt.

Digital isothermal quantification of nucleic acids via simultaneous chemical initiation of recombinase polymerase amplification reactions on SlipChip.

PubMed

Shen, Feng; Davydova, Elena K; Du, Wenbin; Kreutz, Jason E; Piepenburg, Olaf; Ismagilov, Rustem F

2011-05-01

In this paper, digital quantitative detection of nucleic acids was achieved at the single-molecule level by chemical initiation of over one thousand sequence-specific, nanoliter isothermal amplification reactions in parallel. Digital polymerase chain reaction (digital PCR), a method used for quantification of nucleic acids, counts the presence or absence of amplification of individual molecules. However, it still requires temperature cycling, which is undesirable under resource-limited conditions. This makes isothermal methods for nucleic acid amplification, such as recombinase polymerase amplification (RPA), more attractive. A microfluidic digital RPA SlipChip is described here for simultaneous initiation of over one thousand nL-scale RPA reactions by adding a chemical initiator to each reaction compartment with a simple slipping step after instrument-free pipet loading. Two designs of the SlipChip, two-step slipping and one-step slipping, were validated using digital RPA. By using the digital RPA SlipChip, false-positive results from preinitiation of the RPA amplification reaction before incubation were eliminated. End point fluorescence readout was used for "yes or no" digital quantification. The performance of digital RPA in a SlipChip was validated by amplifying and counting single molecules of the target nucleic acid, methicillin-resistant Staphylococcus aureus (MRSA) genomic DNA. The digital RPA on SlipChip was also tolerant to fluctuations of the incubation temperature (37-42 °C), and its performance was comparable to digital PCR on the same SlipChip design. The digital RPA SlipChip provides a simple method to quantify nucleic acids without requiring thermal cycling or kinetic measurements, with potential applications in diagnostics and environmental monitoring under resource-limited settings. The ability to initiate thousands of chemical reactions in parallel on the nanoliter scale using solvent-resistant glass devices is likely to be useful for a broader range of applications.

Digital Isothermal Quantification of Nucleic Acids via Simultaneous Chemical Initiation of Recombinase Polymerase Amplification Reactions on SlipChip

PubMed Central

Shen, Feng; Davydova, Elena K.; Du, Wenbin; Kreutz, Jason E.; Piepenburg, Olaf; Ismagilov, Rustem F.

2011-01-01

In this paper, digital quantitative detection of nucleic acids was achieved at the single-molecule level by chemical initiation of over one thousand sequence-specific, nanoliter, isothermal amplification reactions in parallel. Digital polymerase chain reaction (digital PCR), a method used for quantification of nucleic acids, counts the presence or absence of amplification of individual molecules. However it still requires temperature cycling, which is undesirable under resource-limited conditions. This makes isothermal methods for nucleic acid amplification, such as recombinase polymerase amplification (RPA), more attractive. A microfluidic digital RPA SlipChip is described here for simultaneous initiation of over one thousand nL-scale RPA reactions by adding a chemical initiator to each reaction compartment with a simple slipping step after instrument-free pipette loading. Two designs of the SlipChip, two-step slipping and one-step slipping, were validated using digital RPA. By using the digital RPA SlipChip, false positive results from pre-initiation of the RPA amplification reaction before incubation were eliminated. End-point fluorescence readout was used for “yes or no” digital quantification. The performance of digital RPA in a SlipChip was validated by amplifying and counting single molecules of the target nucleic acid, Methicillin-resistant Staphylococcus aureus (MRSA) genomic DNA. The digital RPA on SlipChip was also tolerant to fluctuations of the incubation temperature (37–42 °C), and its performance was comparable to digital PCR on the same SlipChip design. The digital RPA SlipChip provides a simple method to quantify nucleic acids without requiring thermal cycling or kinetic measurements, with potential applications in diagnostics and environmental monitoring under resource-limited settings. The ability to initiate thousands of chemical reactions in parallel on the nanoliter scale using solvent-resistant glass devices is likely to be useful for a broader range of applications. PMID:21476587

How to Radicalize Graduate Students: Take These Simple Steps to Intimidate, Alienate, and Agitate Your Own Flock of Graduate Students

ERIC Educational Resources Information Center

Steffen, Heather

2011-01-01

As the recession, budget cuts, endowment losses, and Republican governors gut university funding, campuses across the country have become host to occupations, union actions, and demonstrations. One can take steps to intimidate, alienate, and agitate a flock of graduate students. The author discusses steps that offer simple ways to rile up and…

Development and evaluation of an in-house single step loop-mediated isothermal amplification (SS-LAMP) assay for the detection of Mycobacterium tuberculosis complex in sputum samples from Moroccan patients.

PubMed

Bentaleb, El Mehdi; Abid, Mohammed; El Messaoudi, My Driss; Lakssir, Brahim; Ressami, El Mostafa; Amzazi, Saaïd; Sefrioui, Hassan; Ait Benhassou, Hassan

2016-09-27

Tuberculosis (TB) is a major global health problem and remains the leading cause of morbidity and mortality in developing countries. Routinely used TB diagnostic methods, in most endemic areas, are time-consuming, often less-sensitive, expensive and inaccessible to most patients. Therefore, there is an urgent need for the development of early, easy to use and effective diagnosis tools of TB, which can be effectively integrated into resource limited settings, to anticipate the early treatment and limit further spread of the disease. Over the last decade, Loop-mediated isothermal amplification (LAMP) assays have become a powerful tool for rapid diagnosis of infectious diseases because of the simplicity of device requirements. Indeed, LAMP is a simple, quick and cost effective Isothermal Nucleic Acid Amplification diagnostic test (INAAT) that has the potential to be used in TB endemic settings of resource-poor countries. In the present study, we have developed a simple and rapid TB molecular diagnostic test using a Single-Step Loop-mediated isothermal DNA amplification (SS-LAMP) method for the detection of Mycobacterium tuberculosis complex (MTBC) strains, with a simplified sample preparation procedure, eliminating DNA extraction prior to LAMP amplification, DNA initial denaturation and enzymatic inactivation steps during the amplification process. To perform our in-house SS-LAMP assay, a set of six specific primers was specifically designed to recognize eight distinct regions on the MTBC species-specific repetitive insertion sequence 6110 (IS6110). The amplification of the targeted DNA was carried out under isothermal conditions at 65 °C within 1 h. Our protocol was firstly optimized using 60 of confirmed MTBC isolates and a recombinant pGEMeasy-IS6110 vector for sensitivity testing. Thereafter, the assay was evaluated on liquefied sputum specimens collected from 157 Moroccan patients suspected of having TB. Our SS-LAMP developed assay was able to detect MTBC DNA directly from liquefied sputum samples without any prior DNA extraction, denaturation nor the final enzymatic inactivation step. When compared to routinely used Löwenstein Jensen (LJ) Culture method, our SS-LAMP assay is rapid and showed specificity and sensitivity of 99.14 % and 82.93 % respectively which are within the international standards. In addition, the limit of detection of our assay was found to be as little as 10 copies of bacterial DNA. To our knowledge, this is the first study using a single step LAMP (SS-LAMP) procedure as a rapid, easy to perform and cost effective testing for TB early detection. This innovative assay could be suitable for low-income countries with restricted health equipment facilities.

Fiber-integrated refractive index sensor based on a diced Fabry-Perot micro-resonator.

PubMed

Suntsov, Sergiy; Rüter, Christian E; Schipkowski, Tom; Kip, Detlef

2017-11-20

We report on a fiber-integrated refractive index sensor based on a Fabry-Perot micro-resonator fabricated using simple diamond blade dicing of a single-mode step-index fiber. The performance of the device has been tested for the refractive index measurements of sucrose solutions as well as in air. The device shows a sensitivity of 1160 nm/RIU (refractive index unit) at a wavelength of 1.55 μm and a temperature cross-sensitivity of less than 10 -7   RIU/°C. Based on evaluation of the broadband reflection spectra, refractive index steps of 10 -5 of the solutions were accurately measured. The conducted coating of the resonator sidewalls with layers of a high-index material with real-time reflection spectrum monitoring could help to significantly improve the sensor performance.

Environmentally-friendly lithium recycling from a spent organic li-ion battery.

PubMed

Renault, Stéven; Brandell, Daniel; Edström, Kristina

2014-10-01

A simple and straightforward method using non-polluting solvents and a single thermal treatment step at moderate temperature was investigated as an environmentally-friendly process to recycle lithium from organic electrode materials for secondary lithium batteries. This method, highly dependent on the choice of electrolyte, gives up to 99% of sustained capacity for the recycled materials used in a second life-cycle battery when compared with the original. The best results were obtained using a dimethyl carbonate/lithium bis(trifluoromethane sulfonyl) imide electrolyte that does not decompose in presence of water. The process implies a thermal decomposition step at a moderate temperature of the extracted organic material into lithium carbonate, which is then used as a lithiation agent for the preparation of fresh electrode material without loss of lithium. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A simple, enaminone-based approach to some bicyclic pyridazinium tetrafluoroborates

PubMed Central

Josefík, František; Svobodová, Markéta; Bertolasi, Valerio

2013-01-01

Summary Easily obtainable cyclic enaminones (piperidin-2-ylidenealkanones) can be transformed into substituted bicyclic pyridazinium tetrafluoroborates upon treatment with corresponding diazonium salts. The transformation can be performed either in a one-pot way or in a two-step process with the isolation of single azo-coupled enaminone as the intermediate. The former method is superior. Under the optimized conditions, a number of pyridazinium salts substituted with both electron-donating and electron-withdrawing substituents was easily synthesized. A mechanism of the formation of the pyridazinium salts is suggested. A partial drawback is the possibility of the formation of a mixture of products when using a different diazonium salt in each step due to a reversibility of the azo coupling. This can be suppressed by using a more reactive diazonium salt before a less reactive one. PMID:23946844

Texture-based segmentation and analysis of emphysema depicted on CT images

NASA Astrophysics Data System (ADS)

Tan, Jun; Zheng, Bin; Wang, Xingwei; Lederman, Dror; Pu, Jiantao; Sciurba, Frank C.; Gur, David; Leader, J. Ken

2011-03-01

In this study we present a texture-based method of emphysema segmentation depicted on CT examination consisting of two steps. Step 1, a fractal dimension based texture feature extraction is used to initially detect base regions of emphysema. A threshold is applied to the texture result image to obtain initial base regions. Step 2, the base regions are evaluated pixel-by-pixel using a method that considers the variance change incurred by adding a pixel to the base in an effort to refine the boundary of the base regions. Visual inspection revealed a reasonable segmentation of the emphysema regions. There was a strong correlation between lung function (FEV1%, FEV1/FVC, and DLCO%) and fraction of emphysema computed using the texture based method, which were -0.433, -.629, and -0.527, respectively. The texture-based method produced more homogeneous emphysematous regions compared to simple thresholding, especially for large bulla, which can appear as speckled regions in the threshold approach. In the texture-based method, single isolated pixels may be considered as emphysema only if neighboring pixels meet certain criteria, which support the idea that single isolated pixels may not be sufficient evidence that emphysema is present. One of the strength of our complex texture-based approach to emphysema segmentation is that it goes beyond existing approaches that typically extract a single or groups texture features and individually analyze the features. We focus on first identifying potential regions of emphysema and then refining the boundary of the detected regions based on texture patterns.

Compact MEMS external cavity tunable laser with ultra-narrow linewidth for coherent detection.

PubMed

Zhang, Di; Zhao, Jianyi; Yang, Qi; Liu, Wen; Fu, Yanfeng; Li, Chao; Luo, Ming; Hu, Shenglei; Hu, Qianggao; Wang, Lei

2012-08-27

A compact and ultra-narrow linewidth tunable laser with an external cavity based on a simple single-axis-MEMS mirror is presented in this paper. We discuss the simulation of this tunable laser using a two-step hybrid analysis method to obtain an optimal design of the device. A wide wavelength tuning range about 40 nm in C-band with a narrow linewidth of less than 50 kHz and wavelength accuracy of ± 1 GHz over the entire tuning range can be achieved experimentally. We also conduct several experiments under different conditions to test the tunable laser. This device shows an excellent performance in both single-carrier polarization-multiplexed quadrature phase-shift keying (PM-QPSK) and multi-carrier orthogonal frequency division multiplexing (OFDM) coherent systems.

A simple, fast, and inexpensive CTAB-PVP-silica based method for genomic DNA isolation from single, small insect larvae and pupae.

PubMed

Huanca-Mamani, W; Rivera-Cabello, D; Maita-Maita, J

2015-07-17

In this study, we report a modified CTAB-PVP method combined with silicon dioxide (silica) treatment for the extraction of high quality genomic DNA from a single larva or pupa. This method efficiently obtains DNA from small specimens, which is difficult and challenging because of the small amount of starting tissue. Maceration with liquid nitrogen, phenol treatment, and the ethanol precipitation step are eliminated using this methodology. The A260/A280 absorbance ratios of the isolated DNA were approximately 1.8, suggesting that the DNA is pure and can be used for further molecular analysis. The quality of the isolated DNA permits molecular applications and represents a fast, cheap, and effective alternative method for laboratories with low budgets.

Printing 2-dimentional droplet array for single-cell reverse transcription quantitative PCR assay with a microfluidic robot.

PubMed

Zhu, Ying; Zhang, Yun-Xia; Liu, Wen-Wen; Ma, Yan; Fang, Qun; Yao, Bo

2015-04-01

This paper describes a nanoliter droplet array-based single-cell reverse transcription quantitative PCR (RT-qPCR) assay method for quantifying gene expression in individual cells. By sequentially printing nanoliter-scale droplets on microchip using a microfluidic robot, all liquid-handling operations including cell encapsulation, lysis, reverse transcription, and quantitative PCR with real-time fluorescence detection, can be automatically achieved. The inhibition effect of cell suspension buffer on RT-PCR assay was comprehensively studied to achieve high-sensitivity gene quantification. The present system was applied in the quantitative measurement of expression level of mir-122 in single Huh-7 cells. A wide distribution of mir-122 expression in single cells from 3061 copies/cell to 79998 copies/cell was observed, showing a high level of cell heterogeneity. With the advantages of full-automation in liquid-handling, simple system structure, and flexibility in achieving multi-step operations, the present method provides a novel liquid-handling mode for single cell gene expression analysis, and has significant potentials in transcriptional identification and rare cell analysis.

Printing 2-Dimentional Droplet Array for Single-Cell Reverse Transcription Quantitative PCR Assay with a Microfluidic Robot

PubMed Central

Zhu, Ying; Zhang, Yun-Xia; Liu, Wen-Wen; Ma, Yan; Fang, Qun; Yao, Bo

2015-01-01

This paper describes a nanoliter droplet array-based single-cell reverse transcription quantitative PCR (RT-qPCR) assay method for quantifying gene expression in individual cells. By sequentially printing nanoliter-scale droplets on microchip using a microfluidic robot, all liquid-handling operations including cell encapsulation, lysis, reverse transcription, and quantitative PCR with real-time fluorescence detection, can be automatically achieved. The inhibition effect of cell suspension buffer on RT-PCR assay was comprehensively studied to achieve high-sensitivity gene quantification. The present system was applied in the quantitative measurement of expression level of mir-122 in single Huh-7 cells. A wide distribution of mir-122 expression in single cells from 3061 copies/cell to 79998 copies/cell was observed, showing a high level of cell heterogeneity. With the advantages of full-automation in liquid-handling, simple system structure, and flexibility in achieving multi-step operations, the present method provides a novel liquid-handling mode for single cell gene expression analysis, and has significant potentials in transcriptional identification and rare cell analysis. PMID:25828383

Single-step direct fabrication of pillar-on-pore hybrid nanostructures in anodizing aluminum for superior superhydrophobic efficiency.

PubMed

Jeong, Chanyoung; Choi, Chang-Hwan

2012-02-01

Conventional electrochemical anodizing processes of metals such as aluminum typically produce planar and homogeneous nanopore structures. If hydrophobically treated, such 2D planar and interconnected pore structures typically result in lower contact angle and larger contact angle hysteresis than 3D disconnected pillar structures and, hence, exhibit inferior superhydrophobic efficiency. In this study, we demonstrate for the first time that the anodizing parameters can be engineered to design novel pillar-on-pore (POP) hybrid nanostructures directly in a simple one-step fabrication process so that superior surface superhydrophobicity can also be realized effectively from the electrochemical anodization process. On the basis of the characteristic of forming a self-ordered porous morphology in a hexagonal array, the modulation of anodizing voltage and duration enabled the formulation of the hybrid-type nanostructures having controlled pillar morphology on top of a porous layer in both mild and hard anodization modes. The hybrid nanostructures of the anodized metal oxide layer initially enhanced the surface hydrophilicity significantly (i.e., superhydrophilic). However, after a hydrophobic monolayer coating, such hybrid nanostructures then showed superior superhydrophobic nonwetting properties not attainable by the plain nanoporous surfaces produced by conventional anodization conditions. The well-regulated anodization process suggests that electrochemical anodizing can expand its usefulness and efficacy to render various metallic substrates with great superhydrophilicity or -hydrophobicity by directly realizing pillar-like structures on top of a self-ordered nanoporous array through a simple one-step fabrication procedure.

Weakly-coupled 4-mode step-index FMF and demonstration of IM/DD MDM transmission.

PubMed

Hu, Tao; Li, Juhao; Ge, Dawei; Wu, Zhongying; Tian, Yu; Shen, Lei; Liu, Yaping; Chen, Su; Li, Zhengbin; He, Yongqi; Chen, Zhangyuan

2018-04-02

Weakly coupled-mode division multiplexing (MDM) over few-mode fibers (FMF) for short-reach transmission has attracted great interest, which can avoid multiple-input-multiple-output digital signal processing (MIMO-DSP) by greatly suppressing modal crosstalk. In this paper, step-index FMF supporting 4 linearity polarization (LP) modes for MIMO-free transmission is designed and fabricated for the first time, to our knowledge. Modal crosstalk of the fiber is suppressed by increasing the mode effective refractive index differences. The same fabrication method as standard single-mode fiber is adopted so that it is practical and cost-effective. The mode multiplexer/demultiplexer (MUX/DEMUX) consists of cascaded mode-selective couplers (MSCs), which are designed and fabricated by tapering the proposed FMF with single-mode fiber (SMF). The mode MUX and DEMUX achieve very low modal crosstalk not only for the multiplexing/demultiplexing but also for the coupling to/from the FMF. Based on the fabricated FMF and mode MUX/DEMUX, we successfully demonstrate the first simultaneous 4-modes (LP 01 , LP 11 , LP 21 & LP 31 ) 10-km FMF transmission with 10-Gb/s intensity modulation and MIMO-free direct detection (IM/DD). The modal crosstalk of the whole transmission link is successfully suppressed to less than -16.5 dB. The experimental results indicate that FMF with simple step-index structure supporting 4 weakly-coupled modes is feasible.

Simplified Two-Time Step Method for Calculating Combustion Rates and Nitrogen Oxide Emissions for Hydrogen/Air and Hydorgen/Oxygen

NASA Technical Reports Server (NTRS)

Molnar, Melissa; Marek, C. John

2005-01-01

A simplified single rate expression for hydrogen combustion and nitrogen oxide production was developed. Detailed kinetics are predicted for the chemical kinetic times using the complete chemical mechanism over the entire operating space. These times are then correlated to the reactor conditions using an exponential fit. Simple first order reaction expressions are then used to find the conversion in the reactor. The method uses a two-time step kinetic scheme. The first time averaged step is used at the initial times with smaller water concentrations. This gives the average chemical kinetic time as a function of initial overall fuel air ratio, temperature, and pressure. The second instantaneous step is used at higher water concentrations (> 1 x 10(exp -20) moles/cc) in the mixture which gives the chemical kinetic time as a function of the instantaneous fuel and water mole concentrations, pressure and temperature (T4). The simple correlations are then compared to the turbulent mixing times to determine the limiting properties of the reaction. The NASA Glenn GLSENS kinetics code calculates the reaction rates and rate constants for each species in a kinetic scheme for finite kinetic rates. These reaction rates are used to calculate the necessary chemical kinetic times. This time is regressed over the complete initial conditions using the Excel regression routine. Chemical kinetic time equations for H2 and NOx are obtained for H2/air fuel and for the H2/O2. A similar correlation is also developed using data from NASA s Chemical Equilibrium Applications (CEA) code to determine the equilibrium temperature (T4) as a function of overall fuel/air ratio, pressure and initial temperature (T3). High values of the regression coefficient R2 are obtained.

Summary of Simplified Two Time Step Method for Calculating Combustion Rates and Nitrogen Oxide Emissions for Hydrogen/Air and Hydrogen/Oxygen

NASA Technical Reports Server (NTRS)

Marek, C. John; Molnar, Melissa

2005-01-01

A simplified single rate expression for hydrogen combustion and nitrogen oxide production was developed. Detailed kinetics are predicted for the chemical kinetic times using the complete chemical mechanism over the entire operating space. These times are then correlated to the reactor conditions using an exponential fit. Simple first order reaction expressions are then used to find the conversion in the reactor. The method uses a two time step kinetic scheme. The first time averaged step is used at the initial times with smaller water concentrations. This gives the average chemical kinetic time as a function of initial overall fuel air ratio, temperature, and pressure. The second instantaneous step is used at higher water concentrations (greater than l x 10(exp -20)) moles per cc) in the mixture which gives the chemical kinetic time as a function of the instantaneous fuel and water mole concentrations, pressure and temperature (T(sub 4)). The simple correlations are then compared to the turbulent mixing times to determine the limiting properties of the reaction. The NASA Glenn GLSENS kinetics code calculates the reaction rates and rate constants for each species in a kinetic scheme for finite kinetic rates. These reaction rates are used to calculate the necessary chemical kinetic times. This time is regressed over the complete initial conditions using the Excel regression routine. Chemical kinetic time equations for H2 and NOx are obtained for H2/Air fuel and for H2/O2. A similar correlation is also developed using data from NASA's Chemical Equilibrium Applications (CEA) code to determine the equilibrium temperature (T(sub 4)) as a function of overall fuel/air ratio, pressure and initial temperature (T(sub 3)). High values of the regression coefficient R squared are obtained.

Approaches to drug therapy for COPD in Russia: a proposed therapeutic algorithm.

PubMed

Zykov, Kirill A; Ovcharenko, Svetlana I

2017-01-01

Until recently, there have been few clinical algorithms for the management of patients with COPD. Current evidence-based clinical management guidelines can appear to be complex, and they lack clear step-by-step instructions. For these reasons, we chose to create a simple and practical clinical algorithm for the management of patients with COPD, which would be applicable to real-world clinical practice, and which was based on clinical symptoms and spirometric parameters that would take into account the pathophysiological heterogeneity of COPD. This optimized algorithm has two main fields, one for nonspecialist treatment by primary care and general physicians and the other for treatment by specialized pulmonologists. Patients with COPD are treated with long-acting bronchodilators and short-acting drugs on a demand basis. If the forced expiratory volume in one second (FEV 1 ) is ≥50% of predicted and symptoms are mild, treatment with a single long-acting muscarinic antagonist or long-acting beta-agonist is proposed. When FEV 1 is <50% of predicted and/or the COPD assessment test score is ≥10, the use of combined bronchodilators is advised. If there is no response to treatment after three months, referral to a pulmonary specialist is recommended for pathophysiological endotyping: 1) eosinophilic endotype with peripheral blood or sputum eosinophilia >3%; 2) neutrophilic endotype with peripheral blood neutrophilia >60% or green sputum; or 3) pauci-granulocytic endotype. It is hoped that this simple, optimized, step-by-step algorithm will help to individualize the treatment of COPD in real-world clinical practice. This algorithm has yet to be evaluated prospectively or by comparison with other COPD management algorithms, including its effects on patient treatment outcomes. However, it is hoped that this algorithm may be useful in daily clinical practice for physicians treating patients with COPD in Russia.

Audiovisual Fundamentals; Basic Equipment Operation and Simple Materials Production.

ERIC Educational Resources Information Center

Bullard, John R.; Mether, Calvin E.

A guide illustrated with simple sketches explains the functions and step-by-step uses of audiovisual (AV) equipment. Principles of projection, audio, AV equipment, lettering, limited-quantity and quantity duplication, and materials preservation are outlined. Apparatus discussed include overhead, opaque, slide-filmstrip, and multiple-loading slide…

A Practical Model for Forecasting New Freshman Enrollment during the Application Period.

ERIC Educational Resources Information Center

Paulsen, Michael B.

1989-01-01

A simple and effective model for forecasting freshman enrollment during the application period is presented step by step. The model requires minimal and readily available information, uses a simple linear regression analysis on a personal computer, and provides updated monthly forecasts. (MSE)

Electroanalytical applications of screen-printable surfactant-induced sol-gel graphite composites

DOEpatents

Guadalupe, Ana R.; Guo, Yizhu

2001-05-15

A process for preparing sol-gel graphite composite electrodes is presented. This process preferably uses the surfactant bis(2-ethylhexyl) sulfosuccinate (AOT) and eliminates the need for a cosolvent, an acidic catalyst, a cellulose binder and a thermal curing step from prior art processes. Fabrication of screen-printed electrodes by this process provides a simple approach for electroanalytical applications in aqueous and nonaqueous solvents. Examples of applications for such composite electrodes produced from this process include biochemical sensors such as disposable, single-use glucose sensors and ligand modified composite sensors for metal ion sensitive sensors.

2D spatially controlled polymer micro patterning for cellular behavior studies

NASA Astrophysics Data System (ADS)

Dinca, V.; Palla-Papavlu, A.; Paraico, I.; Lippert, T.; Wokaun, A.; Dinescu, M.

2011-04-01

A simple and effective method to functionalize glass surfaces that enable polymer micropatterning and subsequent spatially controlled adhesion of cells is reported in this paper. The method involves the application of laser induced forward transfer (LIFT) to achieve polymer patterning in a single step onto cell repellent substrates (i.e. polyethyleneglycol (PEG)). This approach was used to produce micron-size polyethyleneimine (PEI)-patterns alternating with cell-repellent areas. The focus of this work is the ability of SH-SY5Y human neuroblastoma cells to orient, migrate, and produce organized cellular arrangements on laser generated PEI patterns.

Charge-transfer excited states: Seeking a balanced and efficient wave function ansatz in variational Monte Carlo

DOE PAGES

Blunt, Nick S.; Neuscamman, Eric

2017-11-16

We present a simple and efficient wave function ansatz for the treatment of excited charge-transfer states in real-space quantum Monte Carlo methods. Using the recently-introduced variation-after-response method, this ansatz allows a crucial orbital optimization step to be performed beyond a configuration interaction singles expansion, while only requiring calculation of two Slater determinant objects. As a result, we demonstrate this ansatz for the illustrative example of the stretched LiF molecule, for a range of excited states of formaldehyde, and finally for the more challenging ethylene-tetrafluoroethylene molecule.

Super-resolution imaging applied to moving object tracking

NASA Astrophysics Data System (ADS)

Swalaganata, Galandaru; Ratna Sulistyaningrum, Dwi; Setiyono, Budi

2017-10-01

Moving object tracking in a video is a method used to detect and analyze changes that occur in an object that being observed. Visual quality and the precision of the tracked target are highly wished in modern tracking system. The fact that the tracked object does not always seem clear causes the tracking result less precise. The reasons are low quality video, system noise, small object, and other factors. In order to improve the precision of the tracked object especially for small object, we propose a two step solution that integrates a super-resolution technique into tracking approach. First step is super-resolution imaging applied into frame sequences. This step was done by cropping the frame in several frame or all of frame. Second step is tracking the result of super-resolution images. Super-resolution image is a technique to obtain high-resolution images from low-resolution images. In this research single frame super-resolution technique is proposed for tracking approach. Single frame super-resolution was a kind of super-resolution that it has the advantage of fast computation time. The method used for tracking is Camshift. The advantages of Camshift was simple calculation based on HSV color that use its histogram for some condition and color of the object varies. The computational complexity and large memory requirements required for the implementation of super-resolution and tracking were reduced and the precision of the tracked target was good. Experiment showed that integrate a super-resolution imaging into tracking technique can track the object precisely with various background, shape changes of the object, and in a good light conditions.

Modeling-independent elucidation of inactivation pathways in recombinant and native A-type Kv channels

PubMed Central

Fineberg, Jeffrey D.; Ritter, David M.

2012-01-01

A-type voltage-gated K+ (Kv) channels self-regulate their activity by inactivating directly from the open state (open-state inactivation [OSI]) or by inactivating before they open (closed-state inactivation [CSI]). To determine the inactivation pathways, it is often necessary to apply several pulse protocols, pore blockers, single-channel recording, and kinetic modeling. However, intrinsic hurdles may preclude the standardized application of these methods. Here, we implemented a simple method inspired by earlier studies of Na+ channels to analyze macroscopic inactivation and conclusively deduce the pathways of inactivation of recombinant and native A-type Kv channels. We investigated two distinct A-type Kv channels expressed heterologously (Kv3.4 and Kv4.2 with accessory subunits) and their native counterparts in dorsal root ganglion and cerebellar granule neurons. This approach applies two conventional pulse protocols to examine inactivation induced by (a) a simple step (single-pulse inactivation) and (b) a conditioning step (double-pulse inactivation). Consistent with OSI, the rate of Kv3.4 inactivation (i.e., the negative first derivative of double-pulse inactivation) precisely superimposes on the profile of the Kv3.4 current evoked by a single pulse because the channels must open to inactivate. In contrast, the rate of Kv4.2 inactivation is asynchronous, already changing at earlier times relative to the profile of the Kv4.2 current evoked by a single pulse. Thus, Kv4.2 inactivation occurs uncoupled from channel opening, indicating CSI. Furthermore, the inactivation time constant versus voltage relation of Kv3.4 decreases monotonically with depolarization and levels off, whereas that of Kv4.2 exhibits a J-shape profile. We also manipulated the inactivation phenotype by changing the subunit composition and show how CSI and CSI combined with OSI might affect spiking properties in a full computational model of the hippocampal CA1 neuron. This work unambiguously elucidates contrasting inactivation pathways in neuronal A-type Kv channels and demonstrates how distinct pathways might impact neurophysiological activity. PMID:23109714

Method and apparatus for single-stepping coherence events in a multiprocessor system under software control

DOEpatents

Blumrich, Matthias A.; Salapura, Valentina

2010-11-02

An apparatus and method are disclosed for single-stepping coherence events in a multiprocessor system under software control in order to monitor the behavior of a memory coherence mechanism. Single-stepping coherence events in a multiprocessor system is made possible by adding one or more step registers. By accessing these step registers, one or more coherence requests are processed by the multiprocessor system. The step registers determine if the snoop unit will operate by proceeding in a normal execution mode, or operate in a single-step mode.

A simple protocol for Matrix Assisted Laser Desorption Ionization- time of flight-mass spectrometry (MALDI-TOF-MS) analysis of lipids and proteins in single microsamples of paintings.

PubMed

van der Werf, Inez D; Calvano, Cosima D; Palmisano, Francesco; Sabbatini, Luigia

2012-03-09

A simple protocol, based on Bligh-Dyer (BD) extraction followed by MALDI-TOF-MS analysis, for fast identification of paint binders in single microsamples is proposed. For the first time it is demonstrated that the BD method is effective for the simultaneous extraction of lipids and proteins from complex, and atypical matrices, such as pigmented paint layers. The protocol makes use of an alternative denaturing anionic detergent (RapiGest™) in order to improve efficiency of protein digestion and purification step. Detection of various lipid classes, such as triacylglycerols (TAGs) and phospholipids (PLs), and their oxidation by-products was accomplished, whereas proteins could be identified by peptide mass fingerprinting. The effect of pigments on ageing of lipids and proteins was also investigated. Finally, the proposed protocol was successfully applied to the study of a late-15th century Italian panel painting allowing the identification of various proteinaceous and lipid sections in organic binders, such as egg yolk, egg white, animal glue, casein, and drying oil. Copyright © 2011 Elsevier B.V. All rights reserved.

Control of DNA strand displacement kinetics using toehold exchange.

PubMed

Zhang, David Yu; Winfree, Erik

2009-12-02

DNA is increasingly being used as the engineering material of choice for the construction of nanoscale circuits, structures, and motors. Many of these enzyme-free constructions function by DNA strand displacement reactions. The kinetics of strand displacement can be modulated by toeholds, short single-stranded segments of DNA that colocalize reactant DNA molecules. Recently, the toehold exchange process was introduced as a method for designing fast and reversible strand displacement reactions. Here, we characterize the kinetics of DNA toehold exchange and model it as a three-step process. This model is simple and quantitatively predicts the kinetics of 85 different strand displacement reactions from the DNA sequences. Furthermore, we use toehold exchange to construct a simple catalytic reaction. This work improves the understanding of the kinetics of nucleic acid reactions and will be useful in the rational design of dynamic DNA and RNA circuits and nanodevices.

Comparison study on mechanical properties single step and three step artificial aging on duralium

NASA Astrophysics Data System (ADS)

Tsamroh, Dewi Izzatus; Puspitasari, Poppy; Andoko, Sasongko, M. Ilman N.; Yazirin, Cepi

2017-09-01

Duralium is kind of non-ferro alloy that used widely in industrial. That caused its properties such as mild, high ductility, and resistance from corrosion. This study aimed to know mechanical properties of duralium on single step and three step articial aging process. Mechanical properties that discussed in this study focused on toughness value, tensile strength, and microstructure of duralium. Toughness value of single step artificial aging was 0.082 joule/mm2, and toughness value of three step artificial aging was 0,0721 joule/mm2. Duralium tensile strength of single step artificial aging was 32.36 kgf/mm^2, and duralium tensile strength of three step artificial aging was 32,70 kgf/mm^2. Based on microstructure photo of duralium of single step artificial aging showed that precipitate (θ) was not spreading evenly indicated by black spot which increasing the toughness of material. While microstructure photo of duralium that treated by three step artificial aging showed that it had more precipitate (θ) spread evenly compared with duralium that treated by single step artificial aging.

Simple and rapid quantification of brominated vegetable oil in commercial soft drinks by LC–MS

PubMed Central

Chitranshi, Priyanka; da Costa, Gonçalo Gamboa

2016-01-01

We report here a simple and rapid method for the quantification of brominated vegetable oil (BVO) in soft drinks based upon liquid chromatography–electrospray ionization mass spectrometry. Unlike previously reported methods, this novel method does not require hydrolysis, extraction or derivatization steps, but rather a simple “dilute and shoot” sample preparation. The quantification is conducted by mass spectrometry in selected ion recording mode and a single point standard addition procedure. The method was validated in the range of 5–25 μg/mL BVO, encompassing the legal limit of 15 μg/mL established by the US FDA for fruit-flavored beverages in the US market. The method was characterized by excellent intra- and inter-assay accuracy (97.3–103.4%) and very low imprecision [0.5–3.6% (RSD)]. The direct nature of the quantification, simplicity, and excellent statistical performance of this methodology constitute clear advantages in relation to previously published methods for the analysis of BVO in soft drinks. PMID:27451219

Production of interferon-alpha in high cell density cultures of recombinant Escherichia coli and its single step purification from refolded inclusion body proteins.

PubMed

Babu, K R; Swaminathan, S; Marten, S; Khanna, N; Rinas, U

2000-06-01

Escherichia coli TG1 transformed with a temperature-regulated interferon-alpha expression vector was grown to high cell density in defined medium containing glucose as the sole carbon and energy source, utilizing a simple fed-batch process. Feeding was carried out to achieve an exponential increase in biomass at growth rates which minimized acetate production. Thermal induction of such high cell density cultures resulted in the production of approximately 4 g interferon-alpha/l culture broth. Interferon-alpha was produced exclusively in the form of insoluble inclusion bodies and was solubilized under denaturing conditions, refolded in the presence of arginine and purified to near homogeneity, utilizing single-step ion-exchange chromatography on Q-Sepharose. The yield of purified interferon-alpha was approximately 300 mg/l with respect to the original high cell density culture broth (overall yield of approximately 7.5% active interferon-alpha). The purified recombinant interferon-alpha was found by different criteria to be predominantly monomeric and possessed a specific bioactivity of approximately 2.5 x 10(8) IU/mg based on viral cytopathic assay.

Single-step production of the simvastatin precursor monacolin J by engineering of an industrial strain of Aspergillus terreus.

PubMed

Huang, Xuenian; Liang, Yajing; Yang, Yong; Lu, Xuefeng

2017-07-01

Monacolin J is a key precursor for the synthesis of simvastatin (Zocor), an important drug for treating hypercholesterolemia. Industrially, monacolin J is manufactured through alkaline hydrolysis of lovastatin, a fungal polyketide produced by Aspergillus terreus. Multistep chemical processes for the conversion of lovastatin to simvastatin are laborious, cost expensive and environmentally unfriendly. A biocatalysis process for monacolin J conversion to simvastatin has been developed. However, direct bioproduction of monacolin J has not yet been achieved. Here, we identified a lovastatin hydrolase from Penicillium chrysogenum, which displays a 232-fold higher catalytic efficiency for the in vitro hydrolysis of lovastatin compared to a previously patented hydrolase, but no activity for simvastatin. Furthermore, we showed that an industrial A. terreus strain heterologously expressing this lovastatin hydrolase can produce monacolin J through single-step fermentation with high efficiency, approximately 95% of the biosynthesized lovastatin was hydrolyzed to monacolin J. Our results demonstrate a simple and green technical route for the production of monacolin J, which makes complete bioproduction of the cholesterol-lowering drug simvastatin feasible and promising. Copyright © 2017 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

A highly relevant and efficient single step method for simultaneous depletion and isolation of human regulatory T cells in a clinical setting.

PubMed

Brezar, Vedran; Ruffin, Nicolas; Lévy, Yves; Seddiki, Nabila

2014-09-01

Regulatory T cells (Tregs) are pivotal in preventing autoimmunity. They play a major but still ambiguous role in cancer and viral infections. Functional studies of human Tregs are often hampered by numerous technical difficulties arising from imperfections in isolating and depleting protocols, together with the usual low cell number available from clinical samples. We standardized a simple procedure (Single Step Method, SSM), based on magnetic beads technology, in which both depletion and isolation of human Tregs with high purities are simultaneously achieved. SSM is suitable when using low cell numbers either fresh or frozen from both patients and healthy individuals. It allows simultaneous Tregs isolation and depletion that can be used for further functional work to monitor suppressive function of isolated Tregs (in vitro suppression assay) and also effector IFN-γ responses of Tregs-depleted cell fraction (OX40 assay). To our knowledge, there is no accurate standardized method for Tregs isolation and depletion in a clinical context. SSM could thus be used and easily standardized across different laboratories. Copyright © 2014 Elsevier B.V. All rights reserved.

General solutions for the oxidation kinetics of polymers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gillen, K.T.; Clough, R.L.; Wise, J.

1996-08-01

The simplest general kinetic schemes applicable to the oxidation of polymers are presented, discussed and analyzed in terms of the underlying kinetic assumptions. For the classic basic autoxidation scheme (BAS), which involves three bimolecular termination steps and is applicable mainly to unstabilized polymers, typical assumptions used singly or in groups include (1) long kinetic chain length, (2) a specific ratio of the termination rate constants and (3) insensitivity to the oxygen concentration (e.g., domination by a single termination step). Steady-state solutions for the rate of oxidation are given in terms of one, two, three, or four parameters, corresponding respectively tomore » three, two, one, or zero kinetic assumptions. The recently derived four-parameter solution predicts conditions yielding unusual dependencies of the oxidation rate on oxygen concentration and on initiation rate, as well as conditions leading to some unusual diffusion-limited oxidation profile shapes. For stabilized polymers, unimolecular termination schemes are typically more appropriate than bimolecular. Kinetics incorporating unimolecular termination reactions are shown to result in very simple oxidation expressions which have been experimentally verified for both radiation-initiated oxidation of an EPDM and thermoxidative degradation of nitrile and chloroprene elastomers.« less

DNA assembler, an in vivo genetic method for rapid construction of biochemical pathways

PubMed Central

Shao, Zengyi; Zhao, Hua; Zhao, Huimin

2009-01-01

The assembly of large recombinant DNA encoding a whole biochemical pathway or genome represents a significant challenge. Here, we report a new method, DNA assembler, which allows the assembly of an entire biochemical pathway in a single step via in vivo homologous recombination in Saccharomyces cerevisiae. We show that DNA assembler can rapidly assemble a functional d-xylose utilization pathway (∼9 kb DNA consisting of three genes), a functional zeaxanthin biosynthesis pathway (∼11 kb DNA consisting of five genes) and a functional combined d-xylose utilization and zeaxanthin biosynthesis pathway (∼19 kb consisting of eight genes) with high efficiencies (70–100%) either on a plasmid or on a yeast chromosome. As this new method only requires simple DNA preparation and one-step yeast transformation, it represents a powerful tool in the construction of biochemical pathways for synthetic biology, metabolic engineering and functional genomics studies. PMID:19074487

Learning invariance from natural images inspired by observations in the primary visual cortex.

PubMed

Teichmann, Michael; Wiltschut, Jan; Hamker, Fred

2012-05-01

The human visual system has the remarkable ability to largely recognize objects invariant of their position, rotation, and scale. A good interpretation of neurobiological findings involves a computational model that simulates signal processing of the visual cortex. In part, this is likely achieved step by step from early to late areas of visual perception. While several algorithms have been proposed for learning feature detectors, only few studies at hand cover the issue of biologically plausible learning of such invariance. In this study, a set of Hebbian learning rules based on calcium dynamics and homeostatic regulations of single neurons is proposed. Their performance is verified within a simple model of the primary visual cortex to learn so-called complex cells, based on a sequence of static images. As a result, the learned complex-cell responses are largely invariant to phase and position.

Magnetic Memory from Site Isolated Dy(III) on Silica Materials

PubMed Central

2017-01-01

Achieving magnetic remanence at single isolated metal sites dispersed at the surface of a solid matrix has been envisioned as a key step toward information storage and processing in the smallest unit of matter. Here, we show that isolated Dy(III) sites distributed at the surface of silica nanoparticles, prepared with a simple and scalable two-step process, show magnetic remanence and display a hysteresis loop open at liquid 4He temperature, in contrast to the molecular precursor which does not display any magnetic memory. This singular behavior is achieved through the controlled grafting of a tailored Dy(III) siloxide complex on partially dehydroxylated silica nanoparticles followed by thermal annealing. This approach allows control of the density and the structure of isolated, “bare” Dy(III) sites bound to the silica surface. During the process, all organic fragments are removed, leaving the surface as the sole ligand, promoting magnetic remanence. PMID:28386602

Magnetic memory from site isolated Dy(III) on silica materials

DOE Office of Scientific and Technical Information (OSTI.GOV)

Allouche, Florian; Lapadula, Giuseppe; Siddiqi, Georges

Achieving magnetic remanence at single isolated metal sites dispersed at the surface of a solid matrix has been envisioned as a key step toward information storage and processing in the smallest unit of matter. Here, we show that isolated Dy(III) sites distributed at the surface of silica nanoparticles, prepared with a simple and scalable two-step process, show magnetic remanence and display a hysteresis loop open at liquid 4He temperature, in contrast to the molecular precursor which does not display any magnetic memory. This singular behavior is achieved through the controlled grafting of a tailored Dy(III) siloxide complex on partially dehydroxylatedmore » silica nanoparticles followed by thermal annealing. This approach allows control of the density and the structure of isolated, “bare” Dy(III) sites bound to the silica surface. Throughout the process, all organic fragments are removed, leaving the surface as the sole ligand, promoting magnetic remanence.« less

Magnetic memory from site isolated Dy(III) on silica materials

DOE PAGES

Allouche, Florian; Lapadula, Giuseppe; Siddiqi, Georges; ...

2017-02-22

Achieving magnetic remanence at single isolated metal sites dispersed at the surface of a solid matrix has been envisioned as a key step toward information storage and processing in the smallest unit of matter. Here, we show that isolated Dy(III) sites distributed at the surface of silica nanoparticles, prepared with a simple and scalable two-step process, show magnetic remanence and display a hysteresis loop open at liquid 4He temperature, in contrast to the molecular precursor which does not display any magnetic memory. This singular behavior is achieved through the controlled grafting of a tailored Dy(III) siloxide complex on partially dehydroxylatedmore » silica nanoparticles followed by thermal annealing. This approach allows control of the density and the structure of isolated, “bare” Dy(III) sites bound to the silica surface. Throughout the process, all organic fragments are removed, leaving the surface as the sole ligand, promoting magnetic remanence.« less

Photonic Quantum Networks formed from NV− centers

PubMed Central

Nemoto, Kae; Trupke, Michael; Devitt, Simon J.; Scharfenberger, Burkhard; Buczak, Kathrin; Schmiedmayer, Jörg; Munro, William J.

2016-01-01

In this article we present a simple repeater scheme based on the negatively-charged nitrogen vacancy centre in diamond. Each repeater node is built from modules comprising an optical cavity containing a single NV−, with one nuclear spin from 15N as quantum memory. The module uses only deterministic processes and interactions to achieve high fidelity operations (>99%), and modules are connected by optical fiber. In the repeater node architecture, the processes between modules by photons can be in principle deterministic, however current limitations on optical components lead the processes to be probabilistic but heralded. Our resource-modest repeater architecture contains two modules at each node, and the repeater nodes are then connected by entangled photon pairs. We discuss the performance of such a quantum repeater network with modest resources and then incorporate more resource-intense strategies step by step. Our architecture should allow large-scale quantum information networks with existing or near future technology. PMID:27215433

Photonic Quantum Networks formed from NV(-) centers.

PubMed

Nemoto, Kae; Trupke, Michael; Devitt, Simon J; Scharfenberger, Burkhard; Buczak, Kathrin; Schmiedmayer, Jörg; Munro, William J

2016-05-24

In this article we present a simple repeater scheme based on the negatively-charged nitrogen vacancy centre in diamond. Each repeater node is built from modules comprising an optical cavity containing a single NV(-), with one nuclear spin from (15)N as quantum memory. The module uses only deterministic processes and interactions to achieve high fidelity operations (>99%), and modules are connected by optical fiber. In the repeater node architecture, the processes between modules by photons can be in principle deterministic, however current limitations on optical components lead the processes to be probabilistic but heralded. Our resource-modest repeater architecture contains two modules at each node, and the repeater nodes are then connected by entangled photon pairs. We discuss the performance of such a quantum repeater network with modest resources and then incorporate more resource-intense strategies step by step. Our architecture should allow large-scale quantum information networks with existing or near future technology.

Characterization and multi-step transketolase-ω-transaminase bioconversions in an immobilized enzyme microreactor (IEMR) with packed tube.

PubMed

Halim, Amanatuzzakiah Abdul; Szita, Nicolas; Baganz, Frank

2013-12-01

The concept of de novo metabolic engineering through novel synthetic pathways offers new directions for multi-step enzymatic synthesis of complex molecules. This has been complemented by recent progress in performing enzymatic reactions using immobilized enzyme microreactors (IEMR). This work is concerned with the construction of de novo designed enzyme pathways in a microreactor synthesizing chiral molecules. An interesting compound, commonly used as the building block in several pharmaceutical syntheses, is a single diastereoisomer of 2-amino-1,3,4-butanetriol (ABT). This chiral amino alcohol can be synthesized from simple achiral substrates using two enzymes, transketolase (TK) and transaminase (TAm). Here we describe the development of an IEMR using His6-tagged TK and TAm immobilized onto Ni-NTA agarose beads and packed into tubes to enable multi-step enzyme reactions. The kinetic parameters of both enzymes were first determined using single IEMRs evaluated by a kinetic model developed for packed bed reactors. The Km(app) for both enzymes appeared to be flow rate dependent, while the turnover number kcat was reduced 3 fold compared to solution-phase TK and TAm reactions. For the multi-step enzyme reaction, single IEMRs were cascaded in series, whereby the first enzyme, TK, catalyzed a model reaction of lithium-hydroxypyruvate (HPA) and glycolaldehyde (GA) to L-erythrulose (ERY), and the second unit of the IEMR with immobilized TAm converted ERY into ABT using (S)-α-methylbenzylamine (MBA) as amine donor. With initial 60mM (HPA and GA each) and 6mM (MBA) substrate concentration mixture, the coupled reaction reached approximately 83% conversion in 20 min at the lowest flow rate. The ability to synthesize a chiral pharmaceutical intermediate, ABT in relatively short time proves this IEMR system as a powerful tool for construction and evaluation of de novo pathways as well as for determination of enzyme kinetics. Copyright © 2013 The Authors. Published by Elsevier B.V. All rights reserved.

Rapid prototyping of Fresnel zone plates via direct Ga(+) ion beam lithography for high-resolution X-ray imaging.

PubMed

Keskinbora, Kahraman; Grévent, Corinne; Eigenthaler, Ulrike; Weigand, Markus; Schütz, Gisela

2013-11-26

A significant challenge to the wide utilization of X-ray microscopy lies in the difficulty in fabricating adequate high-resolution optics. To date, electron beam lithography has been the dominant technique for the fabrication of diffractive focusing optics called Fresnel zone plates (FZP), even though this preparation method is usually very complicated and is composed of many fabrication steps. In this work, we demonstrate an alternative method that allows the direct, simple, and fast fabrication of FZPs using focused Ga(+) beam lithography practically, in a single step. This method enabled us to prepare a high-resolution FZP in less than 13 min. The performance of the FZP was evaluated in a scanning transmission soft X-ray microscope where nanostructures as small as sub-29 nm in width were clearly resolved, with an ultimate cutoff resolution of 24.25 nm, demonstrating the highest first-order resolution for any FZP fabricated by the ion beam lithography technique. This rapid and simple fabrication scheme illustrates the capabilities and the potential of direct ion beam lithography (IBL) and is expected to increase the accessibility of high-resolution optics to a wider community of researchers working on soft X-ray and extreme ultraviolet microscopy using synchrotron radiation and advanced laboratory sources.

Reliable and cost effective design of intermetallic Ni2Si nanowires and direct characterization of its mechanical properties

PubMed Central

Han, Seung Zeon; Kang, Joonhee; Kim, Sung-Dae; Choi, Si-Young; Kim, Hyung Giun; Lee, Jehyun; Kim, Kwangho; Lim, Sung Hwan; Han, Byungchan

2015-01-01

We report that a single crystal Ni2Si nanowire (NW) of intermetallic compound can be reliably designed using simple three-step processes: casting a ternary Cu-Ni-Si alloy, nucleate and growth of Ni2Si NWs as embedded in the alloy matrix via designing discontinuous precipitation (DP) of Ni2Si nanoparticles and thermal aging, and finally chemical etching to decouple the Ni2Si NWs from the alloy matrix. By direct application of uniaxial tensile tests to the Ni2Si NW we characterize its mechanical properties, which were rarely reported in previous literatures. Using integrated studies of first principles density functional theory (DFT) calculations, high-resolution transmission electron microscopy (HRTEM), and energy-dispersive X-ray spectroscopy (EDX) we accurately validate the experimental measurements. Our results indicate that our simple three-step method enables to design brittle Ni2Si NW with high tensile strength of 3.0 GPa and elastic modulus of 60.6 GPa. We propose that the systematic methodology pursued in this paper significantly contributes to opening innovative processes to design various kinds of low dimensional nanomaterials leading to advancement of frontiers in nanotechnology and related industry sectors. PMID:26456769

Fast-Rate Capable Electrode Material with Higher Energy Density than LiFePO4: 4.2V LiVPO4F Synthesized by Scalable Single-Step Solid-State Reaction.

PubMed

Kim, Minkyung; Lee, Seongsu; Kang, Byoungwoo

2016-03-01

Use of compounds that contain fluorine (F) as electrode materials in lithium ion batteries has been considered, but synthesizing single-phase samples of these compounds is a difficult task. Here, it is demonstrated that a simple scalable single-step solid-state process with additional fluorine source can obtain highly pure LiVPO 4 F. The resulting material with submicron particles achieves very high rate capability ≈100 mAh g -1 at 60 C-rate (1-min discharge) and even at 200 C-rate (18 s discharge). It retains superior capacity, ≈120 mAh g -1 at 10 C charge/10 C discharge rate (6-min) for 500 cycles with >95% retention efficiency. Furthermore, LiVPO 4 F shows low polarization even at high rates leading to higher operating potential >3.45 V (≈3.6 V at 60 C-rate), so it achieves high energy density. It is demonstrated for the first time that highly pure LiVPO 4 F can achieve high power capability comparable to LiFePO 4 and much higher energy density (≈521 Wh g -1 at 20 C-rate) than LiFePO 4 even without nanostructured particles. LiVPO 4 F can be a real substitute of LiFePO 4.

Crispr-mediated Gene Targeting of Human Induced Pluripotent Stem Cells.

PubMed

Byrne, Susan M; Church, George M

2015-01-01

CRISPR/Cas9 nuclease systems can create double-stranded DNA breaks at specific sequences to efficiently and precisely disrupt, excise, mutate, insert, or replace genes. However, human embryonic stem or induced pluripotent stem cells (iPSCs) are more difficult to transfect and less resilient to DNA damage than immortalized tumor cell lines. Here, we describe an optimized protocol for genome engineering of human iPSCs using a simple transient transfection of plasmids and/or single-stranded oligonucleotides. With this protocol, we achieve transfection efficiencies greater than 60%, with gene disruption efficiencies from 1-25% and gene insertion/replacement efficiencies from 0.5-10% without any further selection or enrichment steps. We also describe how to design and assess optimal sgRNA target sites and donor targeting vectors; cloning individual iPSC by single cell FACS sorting, and genotyping successfully edited cells.

Acetylcholinesterase biosensor based on single-walled carbon nanotubes--Co phtalocyanine for organophosphorus pesticides detection.

PubMed

Ivanov, A N; Younusov, R R; Evtugyn, G A; Arduini, F; Moscone, D; Palleschi, G

2011-07-15

A simple and reliable technique has been developed for the construction of an amperometric acetylcholinesterase biosensor based on screen-printed carbon electrodes. For the first time, one-step modification using single-walled carbon nanotubes and Co phtalocyanine has been proposed to decrease the working potential and to increase the signal of thiocholine oxidation. The biosensor developed made it possible to detect 5-50 ppb of paraoxon and 2-50 ppb of malaoxon with detection limits of 3 and 2 ppb, respectively (incubation 15 min). The biosensor showed high reproducibility when measurements of the substrate and inhibitor were performed (R.S.D. about 1% and 2.5%, respectively). The reliability of the inhibition measurements was confirmed by testing spiked samples of sparkling and tape waters. Copyright © 2011 Elsevier B.V. All rights reserved.

Immunofluorescence-based methods to monitor DNA end resection

PubMed Central

Mukherjee, Bipasha; Tomimatsu, Nozomi; Burma, Sandeep

2017-01-01

Summary Double-strand breaks (DSBs) are the most deleterious amongst all types of DNA damage that can occur in the cell. These breaks arise from both endogenous (for example, DNA replication stress) as well as exogenous insults (for example, ionizing radiation). DSBs are principally repaired by one of two major pathways: non-homologous end joining (NHEJ) or homologous recombination (HR). NHEJ is an error-prone process that can occur in all phases of the cell cycle, while HR is limited to the S and G2 phases of the cell cycle when a sister chromatid is available as a template for error-free repair. The first step in HR is “DNA end resection”, a process during which the broken DNA end is converted into a long stretch of 3′-ended single-stranded DNA (ssDNA). In recent years, DNA end resection has been identified as a pivotal step that controls “repair pathway choice” i.e., the appropriate choice between NHEJ and HR for DSB repair. Therefore, methods to quantitatively or semi-quantitatively assess DNA end resection have gained importance in laboratories working on DNA repair. In this chapter, we describe two simple immunofluorescence-based techniques to monitor DNA end resection in mammalian cells. The first technique involves immuno-detection of Replication Protein A (RPA), a ssDNA-binding protein that binds to resected DNA. The second technique involves labeling of genomic DNA with 5-bromo-2′-deoxyuridine (BrdU) that can be detected by anti-BrdU antibody only after the DNA becomes single stranded due to resection. These methods are not complicated, do not involve sophisticated instrumentation or reporter constructs, and can be applied to most mammalian cell lines, and therefore, should be of broad utility as simple ways of monitoring DNA end resection in vivo. PMID:25804748

Comparison of single-step and two-step purified coagulants from Moringa oleifera seed for turbidity and DOC removal.

PubMed

Sánchez-Martín, J; Ghebremichael, K; Beltrán-Heredia, J

2010-08-01

The coagulant proteins from Moringa oleifera purified with single-step and two-step ion-exchange processes were used for the coagulation of surface water from Meuse river in The Netherlands. The performances of the two purified coagulants and the crude extract were assessed in terms of turbidity and DOC removal. The results indicated that the optimum dosage of the single-step purified coagulant was more than two times higher compared to the two-step purified coagulant in terms of turbidity removal. And the residual DOC in the two-step purified coagulant was lower than in single-step purified coagulant or crude extract. (c) 2010 Elsevier Ltd. All rights reserved.

Factors affecting GEBV accuracy with single-step Bayesian models.

PubMed

Zhou, Lei; Mrode, Raphael; Zhang, Shengli; Zhang, Qin; Li, Bugao; Liu, Jian-Feng

2018-01-01

A single-step approach to obtain genomic prediction was first proposed in 2009. Many studies have investigated the components of GEBV accuracy in genomic selection. However, it is still unclear how the population structure and the relationships between training and validation populations influence GEBV accuracy in terms of single-step analysis. Here, we explored the components of GEBV accuracy in single-step Bayesian analysis with a simulation study. Three scenarios with various numbers of QTL (5, 50, and 500) were simulated. Three models were implemented to analyze the simulated data: single-step genomic best linear unbiased prediction (GBLUP; SSGBLUP), single-step BayesA (SS-BayesA), and single-step BayesB (SS-BayesB). According to our results, GEBV accuracy was influenced by the relationships between the training and validation populations more significantly for ungenotyped animals than for genotyped animals. SS-BayesA/BayesB showed an obvious advantage over SSGBLUP with the scenarios of 5 and 50 QTL. SS-BayesB model obtained the lowest accuracy with the 500 QTL in the simulation. SS-BayesA model was the most efficient and robust considering all QTL scenarios. Generally, both the relationships between training and validation populations and LD between markers and QTL contributed to GEBV accuracy in the single-step analysis, and the advantages of single-step Bayesian models were more apparent when the trait is controlled by fewer QTL.

Improving the Amazonian Hydrologic Cycle in a Coupled Land-Atmosphere, Single Column Model

NASA Astrophysics Data System (ADS)

Harper, A. B.; Denning, S.; Baker, I.; Prihodko, L.; Branson, M.

2006-12-01

We have coupled a land-surface model, the Simple Biosphere Model (SiB3), to a single column of the Colorado State University General Circulation Model (CSU-GCM) in the Amazon River Basin. This is a preliminary step in the broader goal of improved simulation of Basin-wide hydrology. A previous version of the coupled model (SiB2) showed drought and catastrophic dieback of the Amazon rain forest. SiB3 includes updated soil hydrology and root physiology. Our test area for the coupled single column model is near Santarem, Brazil, where measurements from the km 83 flux tower in the Tapajos National Forest can be used to evaluate model output. The model was run for 2001 using NCEP2 Reanalysis as driver data. Preliminary results show that the updated biosphere model coupled to the GCM produces improved simulations of the seasonal cycle of surface water balance and precipitation. Comparisons of the diurnal and seasonal cycles of surface fluxes are also being made.

Single crystal growth from separated educts and its application to lithium transition-metal oxides

PubMed Central

Freund, F.; Williams, S. C.; Johnson, R. D.; Coldea, R.; Gegenwart, P.; Jesche, A.

2016-01-01

Thorough mixing of the starting materials is the first step of a crystal growth procedure. This holds true for almost any standard technique, whereas the intentional separation of educts is considered to be restricted to a very limited number of cases. Here we show that single crystals of α-Li2IrO3 can be grown from separated educts in an open crucible in air. Elemental lithium and iridium are oxidized and transported over a distance of typically one centimeter. In contrast to classical vapor transport, the process is essentially isothermal and a temperature gradient of minor importance. Single crystals grow from an exposed condensation point placed in between the educts. The method has also been applied to the growth of Li2RuO3, Li2PtO3 and β-Li2IrO3. A successful use of this simple and low cost technique for various other materials is anticipated. PMID:27748402

Integrated single-walled carbon nanotube/microfluidic devices for the study of the sensing mechanism of nanotube sensors.

PubMed

Fu, Qiang; Liu, Jie

2005-07-21

A method to fabricate integrated single-walled carbon nanotube/microfluidic devices was developed. This simple process could be used to directly prepare nanotube thin film transistors within the microfluidic channel and to register SWNT devices with the microfludic channel without the need of an additional alignment step. The microfluidic device was designed to have several inlets that deliver multiple liquid flows to a single main channel. The location and width of each flow in the main channel could be controlled by the relative flow rates. This capability enabled us to study the effect of the location and the coverage area of the liquid flow that contained charged molecules on the conduction of the nanotube devices, providing important information on the sensing mechanism of carbon nanotube sensors. The results showed that in a sensor based on a nanotube thin film field effect transistor, the sensing signal came from target molecules absorbed on or around the nanotubes. The effect from adsorption on metal electrodes was weak.

EASY: a simple tool for simultaneously removing background, deadtime and acoustic ringing in quantitative NMR spectroscopy--part I: basic principle and applications.

PubMed

Jaeger, Christian; Hemmann, Felix

2014-01-01

Elimination of Artifacts in NMR SpectroscopY (EASY) is a simple but very effective tool to remove simultaneously any real NMR probe background signal, any spectral distortions due to deadtime ringdown effects and -specifically- severe acoustic ringing artifacts in NMR spectra of low-gamma nuclei. EASY enables and maintains quantitative NMR (qNMR) as only a single pulse (preferably 90°) is used for data acquisition. After the acquisition of the first scan (it contains the wanted NMR signal and the background/deadtime/ringing artifacts) the same experiment is repeated immediately afterwards before the T1 waiting delay. This second scan contains only the background/deadtime/ringing parts. Hence, the simple difference of both yields clean NMR line shapes free of artefacts. In this Part I various examples for complete (1)H, (11)B, (13)C, (19)F probe background removal due to construction parts of the NMR probes are presented. Furthermore, (25)Mg EASY of Mg(OH)2 is presented and this example shows how extremely strong acoustic ringing can be suppressed (more than a factor of 200) such that phase and baseline correction for spectra acquired with a single pulse is no longer a problem. EASY is also a step towards deadtime-free data acquisition as these effects are also canceled completely. EASY can be combined with any other NMR experiment, including 2D NMR, if baseline distortions are a big problem. © 2013 Published by Elsevier Inc.

Step training improves reaction time, gait and balance and reduces falls in older people: a systematic review and meta-analysis.

PubMed

Okubo, Yoshiro; Schoene, Daniel; Lord, Stephen R

2017-04-01

To examine the effects of stepping interventions on fall risk factors and fall incidence in older people. Electronic databases (PubMed, EMBASE, CINAHL, Cochrane, CENTRAL) and reference lists of included articles from inception to March 2015. Randomised (RCT) or clinical controlled trials (CCT) of volitional and reactive stepping interventions that included older (minimum age 60) people providing data on falls or fall risk factors. Meta-analyses of seven RCTs (n=660) showed that the stepping interventions significantly reduced the rate of falls (rate ratio=0.48, 95% CI 0.36 to 0.65, p<0.0001, I 2 =0%) and the proportion of fallers (risk ratio=0.51, 95% CI 0.38 to 0.68, p<0.0001, I 2 =0%). Subgroup analyses stratified by reactive and volitional stepping interventions revealed a similar efficacy for rate of falls and proportion of fallers. A meta-analysis of two RCTs (n=62) showed that stepping interventions significantly reduced laboratory-induced falls, and meta-analysis findings of up to five RCTs and CCTs (n=36-416) revealed that stepping interventions significantly improved simple and choice stepping reaction time, single leg stance, timed up and go performance (p<0.05), but not measures of strength. The findings indicate that both reactive and volitional stepping interventions reduce falls among older adults by approximately 50%. This clinically significant reduction may be due to improvements in reaction time, gait, balance and balance recovery but not in strength. Further high-quality studies aimed at maximising the effectiveness and feasibility of stepping interventions are required. CRD42015017357. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

A simplified bioprocess for human alpha-fetoprotein production from inclusion bodies.

PubMed

Leong, Susanna S J; Middelberg, Anton P J

2007-05-01

A simple and effective Escherichia coli (E. coli) bioprocess is demonstrated for the preparation of recombinant human alpha-fetoprotein (rhAFP), a pharmaceutically promising protein that has important immunomodulatory functions. The new rhAFP process employs only unit operations that are easy to scale and validate, and reduces the complexity embedded in existing inclusion body processing methods. A key requirement in the establishment of this process was the attainment of high purity rhAFP prior to protein refolding because (i) rhAFP binds easily to hydrophobic contaminants once refolded, and (ii) rhAFP aggregates during renaturation, in a contaminant- dependent way. In this work, direct protein extraction from cell suspension was coupled with a DNA precipitation-centrifugation step prior to purification using two simple chromatographic steps. Refolding was conducted using a single-step, redox-optimized dilution refolding protocol, with refolding success determined by reversed phase HPLC analysis, ELISA, and circular dichroism spectroscopy. Quantitation of DNA and protein contaminant loads after each unit operation showed that contaminant levels were reduced to levels comparable to traditional flowsheets. Protein microchemical modification due to carbamylation in this urea-based process was identified and minimized, yielding a final refolded and purified product that was significantly purified from carbamylated variants. Importantly, this work conclusively demonstrates, for the first time, that a chemical extraction process can substitute the more complex traditional inclusion body processing flowsheet, without compromising product purity and yield. This highly intensified and simplified process is expected to be of general utility for the preparation of other therapeutic candidates expressed as inclusion bodies. (c) 2006 Wiley Periodicals, Inc.

Adsorption dynamics of methyl violet onto granulated mesoporous carbon: Facile synthesis and adsorption kinetics.

PubMed

Kim, Yohan; Bae, Jiyeol; Park, Hosik; Suh, Jeong-Kwon; You, Young-Woo; Choi, Heechul

2016-09-15

A new and facile one-step synthesis method for preparing granulated mesoporous carbon (GMC) with three-dimensional spherical mesoporous symmetry is prepared to remove large molecular weight organic compounds in aqueous phase. GMC is synthesized in a single step using as-synthesized mesoporous carbon particles and organic binders through a simple and economical synthesis approach involving a simultaneous calcination and carbonization process. Characterization results obtained from SEM, XRD, as well as surface and porosity analysis indicate that the synthesized GMC has similar physical properties to those of the powdered mesoporous carbon and maintains the Brunauer-Emmett-Teller (BET) surface area and pore volume because the new synthesis method prevents the collapse of the pores during the granulation process. Batch adsorption experiments revealed GMC showed a substantial adsorption capacity (202.8 mg/g) for the removal of methyl violet as a target large molecular contaminant in aqueous phase. The mechanisms and dynamics modeling of GMC adsorption were also fully examined, which revealed that surface diffusion was rate limiting step on adsorption process of GMC. Adsorption kinetics of GMC enables 3 times faster than that of granular activated carbon in terms of surface diffusion coefficient. This is the first study, to the best of our knowledge, to synthesize GMC as an adsorbent for water purification by using facile granulation method and to investigate the adsorption kinetics and characteristics of GMC. This study introduces a new and simple method for the synthesis of GMC and reveals its adsorption characteristics for large molecular compounds in a water treatment. Copyright © 2016 Elsevier Ltd. All rights reserved.

Simple webs of natural environment theme as a result of sharing in science teacher training

NASA Astrophysics Data System (ADS)

Tapilouw, M. C.; Firman, H.; Redjeki, S.; Chandra, D. T.

2018-03-01

Thematic learning is one type of integrated science (Biology, Physics, Chemistry and Earth Science) in Science Education. This study is concerning about simple webs of natural environment theme in science learning, as one of training material in science teacher training program. Making simple web is a goal of first step in teacher training program. Every group explain their web illustration to other group. Twenty Junior High School science teacher above one education foundation participate in science teacher training program. In order to gather simple webs, sharing method was used in this first step of science teacher training. The result of this study is five different simple web of natural environment themes. These webs represent science learning in class VII/Semester I, class VII/Semester II, Class VIII, Class IX/Semester I, Class IX/Semester II based on basic competency in National Curriculum 2013. Each group discussed web of natural environment theme based on their learning experience in real class which basic competency and subject matters are linked with natural environment theme. As a conclusion, simple webs are potential to develop in the next step of science teacher training program and to be implemented in real class.

How Long is my Toilet Roll?--A Simple Exercise in Mathematical Modelling

ERIC Educational Resources Information Center

Johnston, Peter R.

2013-01-01

The simple question of how much paper is left on my toilet roll is studied from a mathematical modelling perspective. As is typical with applied mathematics, models of increasing complexity are introduced and solved. Solutions produced at each step are compared with the solution from the previous step. This process exposes students to the typical…

Single step modified ink staining for Tzanck test: quick detection of herpetic giant cells in Tzanck smear.

PubMed

Mizutani, Hitoshi; Akeda, Tomoko; Yamanaka, Kei-Ichi; Isoda, Kenichi; Gabazza, Esteban C

2012-02-01

Tzanck test has been recently re-evaluated as a method for the diagnosis of herpes virus infection. Giemsa staining for the Tzanck test is time-consuming and laborious. There is a need to develop simple and quick staining methods for bedside diagnosis of this disease. We report a single step and quick method for staining herpes giant cells in Tzanck smears using routinely available inks and physiological saline. A keratinocyte cell line (HaCaT) was cultured on a slide glass and stained with various commercially available blue, blue-black and black inks serially diluted with physiological saline. Clinical smear samples from herpes lesions were also stained with these solutions without specific pretreatment. The nuclei of HaCaT were clearly stained showing high contrast with the cytoplasm using 5% Parker-Quink blue-black ink saline solution. Concentration of ink solution higher or lower than 5% resulted in less contrast. Blue or black inks or other manufacturers' inks can also be used, but staining of the cultured keratinocytes was less clear. Smear of clinical samples from herpes lesions were also stained with 5% ink solution. The nuclei of the multinucleated giant cells were clearly stained, and the sample could be immediately used for microscopic examination. One step staining of Tzanck smear using this diluted ink solution is an inexpensive and a convenient bedside diagnostic tool for the dermatologist. © 2011 Japanese Dermatological Association.

Single-step generation of gene knockout-rescue system in pluripotent stem cells by promoter insertion with CRISPR/Cas9.

PubMed

Matsunaga, Taichi; Yamashita, Jun K

2014-02-07

Specific gene knockout and rescue experiments are powerful tools in developmental and stem cell biology. Nevertheless, the experiments require multiple steps of molecular manipulation for gene knockout and subsequent rescue procedures. Here we report an efficient and single step strategy to generate gene knockout-rescue system in pluripotent stem cells by promoter insertion with CRISPR/Cas9 genome editing technology. We inserted a tetracycline-regulated inducible gene promoter (tet-OFF/TRE-CMV) upstream of the endogenous promoter region of vascular endothelial growth factor receptor 2 (VEGFR2/Flk1) gene, an essential gene for endothelial cell (EC) differentiation, in mouse embryonic stem cells (ESCs) with homologous recombination. Both homo- and hetero-inserted clones were efficiently obtained through a simple selection with a drug-resistant gene. The insertion of TRE-CMV promoter disrupted endogenous Flk1 expression, resulting in null mutation in homo-inserted clones. When the inserted TRE-CMV promoter was activated with doxycycline (Dox) depletion, Flk1 expression was sufficiently recovered from the downstream genomic Flk1 gene. Whereas EC differentiation was almost completely perturbed in homo-inserted clones, Flk1 rescue with TRE-CMV promoter activation restored EC appearance, indicating that phenotypic changes in EC differentiation can be successfully reproduced with this knockout-rescue system. Thus, this promoter insertion strategy with CRISPR/Cas9 would be a novel attractive method for knockout-rescue experiments. Copyright © 2014 Elsevier Inc. All rights reserved.

Probing static disorder in Arrhenius kinetics by single-molecule force spectroscopy.

PubMed

Kuo, Tzu-Ling; Garcia-Manyes, Sergi; Li, Jingyuan; Barel, Itay; Lu, Hui; Berne, Bruce J; Urbakh, Michael; Klafter, Joseph; Fernández, Julio M

2010-06-22

The widely used Arrhenius equation describes the kinetics of simple two-state reactions, with the implicit assumption of a single transition state with a well-defined activation energy barrier DeltaE, as the rate-limiting step. However, it has become increasingly clear that the saddle point of the free-energy surface in most reactions is populated by ensembles of conformations, leading to nonexponential kinetics. Here we present a theory that generalizes the Arrhenius equation to include static disorder of conformational degrees of freedom as a function of an external perturbation to fully account for a diverse set of transition states. The effect of a perturbation on static disorder is best examined at the single-molecule level. Here we use force-clamp spectroscopy to study the nonexponential kinetics of single ubiquitin proteins unfolding under force. We find that the measured variance in DeltaE shows both force-dependent and independent components, where the force-dependent component scales with F(2), in excellent agreement with our theory. Our study illustrates a novel adaptation of the classical Arrhenius equation that accounts for the microscopic origins of nonexponential kinetics, which are essential in understanding the rapidly growing body of single-molecule data.

Simple method for the generation of multiple homogeneous field volumes inside the bore of superconducting magnets.

PubMed

Chou, Ching-Yu; Ferrage, Fabien; Aubert, Guy; Sakellariou, Dimitris

2015-07-17

Standard Magnetic Resonance magnets produce a single homogeneous field volume, where the analysis is performed. Nonetheless, several modern applications could benefit from the generation of multiple homogeneous field volumes along the axis and inside the bore of the magnet. In this communication, we propose a straightforward method using a combination of ring structures of permanent magnets in order to cancel the gradient of the stray field in a series of distinct volumes. These concepts were demonstrated numerically on an experimentally measured magnetic field profile. We discuss advantages and limitations of our method and present the key steps required for an experimental validation.

Preparation of Ultraviolet Curing Type Silicone Rubbers Containing Mesoporous Silica Fillers.

PubMed

Abdullah, Nawfel; Hossain, Md Shahriar A; Fatehmulla, Amanullah; Farooq, Wazirzada Aslam; Islam, Md Tofazzal; Miyamoto, Nobuyoshi; Bando, Yoshio; Kamachi, Yuichiro; Malgras, Victor; Yamauchi, Yusuke; Suzuki, Norihiro

2018-01-01

Here we have been focusing on mesoporous silica (MPS) as inorganic filler material to improve the mechanical strength of silicone rubbers. The MPS particles are more effective in reducing the coefficient of thermal expansion (CTE) and hardening silicone rubber composites when compared to commercially available nonporous silica particles. In this study, we utilize ultraviolet curing type silicone rubbers and prepare MPS composites according to a simple single-step method. From an industrial viewpoint, simplifying the fabrication processes is critical. The thermal stability and mechanical strength are examined in detail in order to showcase the effectiveness of MPS particles as filler materials.

Method for making electro-fluidic connections in microfluidic devices

DOEpatents

Frye-Mason, Gregory C.; Martinez, David; Manginell, Ronald P.; Heller, Edwin J.; Chanchani, Rajen

2004-08-10

A method for forming electro-fluidic interconnections in microfluidic devices comprises forming an electrical connection between matching bond pads on a die containing an active electrical element and a microfluidic substrate and forming a fluidic seal ring that circumscribes the active electrical element and a fluidic feedthrough. Preferably, the electrical connection and the seal ring are formed in a single bonding step. The simple method is particularly useful for chemical microanalytical systems wherein a plurality of microanalytical components, such as a chemical preconcentrator, a gas chromatography column, and a surface acoustic wave detector, are fluidically interconnected on a hybrid microfluidic substrate having electrical connection to external support electronics.

Combining two-directional synthesis and tandem reactions. Part 21: Exploitation of a dimeric macrocycle for chain terminus differentiation and synthesis of an sp(3)-rich library.

PubMed

Storr, Thomas E; Cully, Sarah J; Rawling, Michael J; Lewis, William; Hamza, Daniel; Jones, Geraint; Stockman, Robert A

2015-06-01

The application of a tandem condensation/cyclisation/[3+2]-cycloaddition/elimination reaction gives an sp(3)-rich tricyclic pyrazoline scaffold with two ethyl esters in a single step from a simple linear starting material. The successive hydrolysis and cyclisation (with Boc anhydride) of these 3-dimensional architectures, generates unprecedented 16-membered macrocyclic bisanhydrides (characterised by XRD). Selective amidations could then be achieved by ring opening with a primary amine followed by HATU-promoted amide coupling to yield an sp(3)-rich natural product-like library. Copyright © 2015 Elsevier Ltd. All rights reserved.

Antibiotic Combinations That Enable One-Step, Targeted Mutagenesis of Chromosomal Genes.

PubMed

Lee, Wonsik; Do, Truc; Zhang, Ge; Kahne, Daniel; Meredith, Timothy C; Walker, Suzanne

2018-06-08

Targeted modification of bacterial chromosomes is necessary to understand new drug targets, investigate virulence factors, elucidate cell physiology, and validate results of -omics-based approaches. For some bacteria, reverse genetics remains a major bottleneck to progress in research. Here, we describe a compound-centric strategy that combines new negative selection markers with known positive selection markers to achieve simple, efficient one-step genome engineering of bacterial chromosomes. The method was inspired by the observation that certain nonessential metabolic pathways contain essential late steps, suggesting that antibiotics targeting a late step can be used to select for the absence of genes that control flux into the pathway. Guided by this hypothesis, we have identified antibiotic/counterselectable markers to accelerate reverse engineering of two increasingly antibiotic-resistant pathogens, Staphylococcus aureus and Acinetobacter baumannii. For S. aureus, we used wall teichoic acid biosynthesis inhibitors to select for the absence of tarO and for A. baumannii, we used colistin to select for the absence of lpxC. We have obtained desired gene deletions, gene fusions, and promoter swaps in a single plating step with perfect efficiency. Our method can also be adapted to generate markerless deletions of genes using FLP recombinase. The tools described here will accelerate research on two important pathogens, and the concept we outline can be readily adapted to any organism for which a suitable target pathway can be identified.

A novel and simple test of gait adaptability predicts gold standard measures of functional mobility in stroke survivors.

PubMed

Hollands, K L; Pelton, T A; van der Veen, S; Alharbi, S; Hollands, M A

2016-01-01

Although there is evidence that stroke survivors have reduced gait adaptability, the underlying mechanisms and the relationship to functional recovery are largely unknown. We explored the relationships between walking adaptability and clinical measures of balance, motor recovery and functional ability in stroke survivors. Stroke survivors (n=42) stepped to targets, on a 6m walkway, placed to elicit step lengthening, shortening and narrowing on paretic and non-paretic sides. The number of targets missed during six walks and target stepping speed was recorded. Fugl-Meyer (FM), Berg Balance Scale (BBS), self-selected walking speed (SWWS) and single support (SS) and step length (SL) symmetry (using GaitRite when not walking to targets) were also assessed. Stepwise multiple-linear regression was used to model the relationships between: total targets missed, number missed with paretic and non-paretic legs, target stepping speed, and each clinical measure. Regression revealed a significant model for each outcome variable that included only one independent variable. Targets missed by the paretic limb, was a significant predictor of FM (F(1,40)=6.54, p=0.014,). Speed of target stepping was a significant predictor of each of BBS (F(1,40)=26.36, p<0.0001), SSWS (F(1,40)=37.00, p<0.0001). No variables were significant predictors of SL or SS asymmetry. Speed of target stepping was significantly predictive of BBS and SSWS and paretic targets missed predicted FM, suggesting that fast target stepping requires good balance and accurate stepping demands good paretic leg function. The relationships between these parameters indicate gait adaptability is a clinically meaningful target for measurement and treatment of functionally adaptive walking ability in stroke survivors. Copyright © 2015 Elsevier B.V. All rights reserved.

Efficient, ultra-high-affinity chromatography in a one-step purification of complex proteins

PubMed Central

Vassylyeva, Marina N.; Klyuyev, Sergiy; Vassylyev, Alexey D.; Wesson, Hunter; Zhang, Zhuo; Renfrow, Matthew B.; Wang, Hengbin; Higgins, N. Patrick; Chow, Louise T.; Vassylyev, Dmitry G.

2017-01-01

Protein purification is an essential primary step in numerous biological studies. It is particularly significant for the rapidly emerging high-throughput fields, such as proteomics, interactomics, and drug discovery. Moreover, purifications for structural and industrial applications should meet the requirement of high yield, high purity, and high activity (HHH). It is, therefore, highly desirable to have an efficient purification system with a potential to meet the HHH benchmark in a single step. Here, we report a chromatographic technology based on the ultra-high-affinity (Kd ∼ 10−14–10−17 M) complex between the Colicin E7 DNase (CE7) and its inhibitor, Immunity protein 7 (Im7). For this application, we mutated CE7 to create a CL7 tag, which retained the full binding affinity to Im7 but was inactivated as a DNase. To achieve high capacity, we developed a protocol for a large-scale production and highly specific immobilization of Im7 to a solid support. We demonstrated its utility with one-step HHH purification of a wide range of traditionally challenging biological molecules, including eukaryotic, membrane, toxic, and multisubunit DNA/RNA-binding proteins. The system is simple, reusable, and also applicable to pulldown and kinetic activity/binding assays. PMID:28607052

Effect of reaction-step-size noise on the switching dynamics of stochastic populations

NASA Astrophysics Data System (ADS)

Be'er, Shay; Heller-Algazi, Metar; Assaf, Michael

2016-05-01

In genetic circuits, when the messenger RNA lifetime is short compared to the cell cycle, proteins are produced in geometrically distributed bursts, which greatly affects the cellular switching dynamics between different metastable phenotypic states. Motivated by this scenario, we study a general problem of switching or escape in stochastic populations, where influx of particles occurs in groups or bursts, sampled from an arbitrary distribution. The fact that the step size of the influx reaction is a priori unknown and, in general, may fluctuate in time with a given correlation time and statistics, introduces an additional nondemographic reaction-step-size noise into the system. Employing the probability-generating function technique in conjunction with Hamiltonian formulation, we are able to map the problem in the leading order onto solving a stationary Hamilton-Jacobi equation. We show that compared to the "usual case" of single-step influx, bursty influx exponentially decreases the population's mean escape time from its long-lived metastable state. In particular, close to bifurcation we find a simple analytical expression for the mean escape time which solely depends on the mean and variance of the burst-size distribution. Our results are demonstrated on several realistic distributions and compare well with numerical Monte Carlo simulations.

Kinetics of inactivation and dilution effects on the mass balance of fungal phytopathogens in anaerobic digesters.

PubMed

Plöchl, Matthias; Heiermann, Monika; Rodemann, Bernd; Bandte, Martina; Büttner, Carmen

2014-01-15

Knowledge of fate and behavior of plant pathogens in the biogas production chain is limited and hampers the estimation and evaluation of the potential phytosanitary risk if digestate is spread on arable land as a fertilizer. Therefore, simulation is an appropriate tool to demonstrate the effects which influence the steady state of pathogen infected plant material in both digesters and digestate. Simple approaches of kinetics of inactivation and mass balances of infected material were carried out considering single-step as well as two-step digestion. The simulation revealed a very fast to fast reduction of infected material after a singular feeding, reaching a cutback to less than 1% of input within 4 days even for D90-values of 68 h. Steady state mass balances below input rate could be calculated with D90-values of less than 2 h at a continuous hourly feeding. At higher D90-values steady state mass balances exceed the input rate but are still clearly below the sum of input mass. Dilution further decreases mass balances to values 10(-5) to 10(-6) Mg m(-3) for first-step digestion and 10(-8) to 10(-9) for second-step. Copyright © 2013 Elsevier Ltd. All rights reserved.

An intelligent artificial throat with sound-sensing ability based on laser induced graphene

PubMed Central

Tao, Lu-Qi; Tian, He; Liu, Ying; Ju, Zhen-Yi; Pang, Yu; Chen, Yuan-Quan; Wang, Dan-Yang; Tian, Xiang-Guang; Yan, Jun-Chao; Deng, Ning-Qin; Yang, Yi; Ren, Tian-Ling

2017-01-01

Traditional sound sources and sound detectors are usually independent and discrete in the human hearing range. To minimize the device size and integrate it with wearable electronics, there is an urgent requirement of realizing the functional integration of generating and detecting sound in a single device. Here we show an intelligent laser-induced graphene artificial throat, which can not only generate sound but also detect sound in a single device. More importantly, the intelligent artificial throat will significantly assist for the disabled, because the simple throat vibrations such as hum, cough and scream with different intensity or frequency from a mute person can be detected and converted into controllable sounds. Furthermore, the laser-induced graphene artificial throat has the advantage of one-step fabrication, high efficiency, excellent flexibility and low cost, and it will open practical applications in voice control, wearable electronics and many other areas. PMID:28232739

An intelligent artificial throat with sound-sensing ability based on laser induced graphene.

PubMed

Tao, Lu-Qi; Tian, He; Liu, Ying; Ju, Zhen-Yi; Pang, Yu; Chen, Yuan-Quan; Wang, Dan-Yang; Tian, Xiang-Guang; Yan, Jun-Chao; Deng, Ning-Qin; Yang, Yi; Ren, Tian-Ling

2017-02-24

Traditional sound sources and sound detectors are usually independent and discrete in the human hearing range. To minimize the device size and integrate it with wearable electronics, there is an urgent requirement of realizing the functional integration of generating and detecting sound in a single device. Here we show an intelligent laser-induced graphene artificial throat, which can not only generate sound but also detect sound in a single device. More importantly, the intelligent artificial throat will significantly assist for the disabled, because the simple throat vibrations such as hum, cough and scream with different intensity or frequency from a mute person can be detected and converted into controllable sounds. Furthermore, the laser-induced graphene artificial throat has the advantage of one-step fabrication, high efficiency, excellent flexibility and low cost, and it will open practical applications in voice control, wearable electronics and many other areas.

Electrochemical Detection in Stacked Paper Networks.

PubMed

Liu, Xiyuan; Lillehoj, Peter B

2015-08-01

Paper-based electrochemical biosensors are a promising technology that enables rapid, quantitative measurements on an inexpensive platform. However, the control of liquids in paper networks is generally limited to a single sample delivery step. Here, we propose a simple method to automate the loading and delivery of liquid samples to sensing electrodes on paper networks by stacking multiple layers of paper. Using these stacked paper devices (SPDs), we demonstrate a unique strategy to fully immerse planar electrodes by aqueous liquids via capillary flow. Amperometric measurements of xanthine oxidase revealed that electrochemical sensors on four-layer SPDs generated detection signals up to 75% higher compared with those on single-layer paper devices. Furthermore, measurements could be performed with minimal user involvement and completed within 30 min. Due to its simplicity, enhanced automation, and capability for quantitative measurements, stacked paper electrochemical biosensors can be useful tools for point-of-care testing in resource-limited settings. © 2015 Society for Laboratory Automation and Screening.

Surface-enhanced Raman scattering from graphene covered gold nanocap arrays

NASA Astrophysics Data System (ADS)

Long, Kailin; Luo, Xiaoguang; Nan, Haiyan; Du, Deyang; Zhao, Weiwei; Ni, Zhenhua; Qiu, Teng

2013-11-01

This work reports an efficient method to fabricate large-area flexible substrates for surface enhanced Raman scattering (SERS) application. Our technique is based on a single-step direct imprint process via porous anodic alumina stamps. Periodic hexagonal arrangements of porous anodic alumina stamps are transferred to the polyethylene terephthalate substrates by mechanically printing process. Printed nanocaps will turn into "hot spots" for electromagnetic enhancement with a deposited gold film by high vacuum evaporation. The gaps between the nanocaps are controllable with a tight correspondence to the thickness of the deposited gold, which dramatically influence the enhancement factor. After covered with a single-layer graphene sheet, the gold nanocap substrate can be further optimized with an extra enhancement of Raman signals, and it is available for the trace detection of probe molecules. This convenient, simple, and low-cost method of making flexible SERS-active substrates potentially opens a way towards biochemical analysis and disease detection.

An intelligent artificial throat with sound-sensing ability based on laser induced graphene

NASA Astrophysics Data System (ADS)

Tao, Lu-Qi; Tian, He; Liu, Ying; Ju, Zhen-Yi; Pang, Yu; Chen, Yuan-Quan; Wang, Dan-Yang; Tian, Xiang-Guang; Yan, Jun-Chao; Deng, Ning-Qin; Yang, Yi; Ren, Tian-Ling

2017-02-01

Traditional sound sources and sound detectors are usually independent and discrete in the human hearing range. To minimize the device size and integrate it with wearable electronics, there is an urgent requirement of realizing the functional integration of generating and detecting sound in a single device. Here we show an intelligent laser-induced graphene artificial throat, which can not only generate sound but also detect sound in a single device. More importantly, the intelligent artificial throat will significantly assist for the disabled, because the simple throat vibrations such as hum, cough and scream with different intensity or frequency from a mute person can be detected and converted into controllable sounds. Furthermore, the laser-induced graphene artificial throat has the advantage of one-step fabrication, high efficiency, excellent flexibility and low cost, and it will open practical applications in voice control, wearable electronics and many other areas.

Vacuum-free laminated top electrode with conductive tapes for scalable manufacturing of efficient perovskite solar cells

DOE PAGES

Shao, Yuchuan; Wang, Qi; Dong, Qingfeng; ...

2015-06-25

The efficiency of organometal trihalide perovskites (OTP) solar cells have reached that parity of single crystal silicon, and its nature abundant raw material and solution-process capability promise a bright future for commercialization. However, the vacuum based techniques for metal electrode deposition and additional encapsulation layer increase the cost of the perovskite solar cells dramatically and impede their commercialization process. Here, we report a vacuum-free low temperature lamination technique to fabricate the top electrode by commercial conductive tapes (C-tape). The simple fabrication method yields good quality contact and high efficiency device of 12.7%. The C-tapes also encapsulated the devices effectively, resultingmore » in greatly improved device stability. As a result, the combination of lamination of electrodes and encapsulation layers into a single step significantly reduce the cost of device fabrication.« less

A curve fitting method for extrinsic camera calibration from a single image of a cylindrical object

NASA Astrophysics Data System (ADS)

Winkler, A. W.; Zagar, B. G.

2013-08-01

An important step in the process of optical steel coil quality assurance is to measure the proportions of width and radius of steel coils as well as the relative position and orientation of the camera. This work attempts to estimate these extrinsic parameters from single images by using the cylindrical coil itself as the calibration target. Therefore, an adaptive least-squares algorithm is applied to fit parametrized curves to the detected true coil outline in the acquisition. The employed model allows for strictly separating the intrinsic and the extrinsic parameters. Thus, the intrinsic camera parameters can be calibrated beforehand using available calibration software. Furthermore, a way to segment the true coil outline in the acquired images is motivated. The proposed optimization method yields highly accurate results and can be generalized even to measure other solids which cannot be characterized by the identification of simple geometric primitives.

Reflection symmetry detection using locally affine invariant edge correspondence.

PubMed

Wang, Zhaozhong; Tang, Zesheng; Zhang, Xiao

2015-04-01

Reflection symmetry detection receives increasing attentions in recent years. The state-of-the-art algorithms mainly use the matching of intensity-based features (such as the SIFT) within a single image to find symmetry axes. This paper proposes a novel approach by establishing the correspondence of locally affine invariant edge-based features, which are superior to the intensity based in the aspects that it is insensitive to illumination variations, and applicable to textureless objects. The locally affine invariance is achieved by simple linear algebra for efficient and robust computations, making the algorithm suitable for detections under object distortions like perspective projection. Commonly used edge detectors and a voting process are, respectively, used before and after the edge description and matching steps to form a complete reflection detection pipeline. Experiments are performed using synthetic and real-world images with both multiple and single reflection symmetry axis. The test results are compared with existing algorithms to validate the proposed method.

Raman and dielectric studies of GdMnO3 bulk ceramics synthesized from nano powders

NASA Astrophysics Data System (ADS)

Samantaray, S.; Mishra, D. K.; Roul, B. K.

2017-05-01

Nanocrystalline GdMnO3 (GMO) powders has been synthesized by a simple chemical route i. e. pyrophoric reaction technique and then sintered in the form of bulk pellet at 850°C for 24 hours by adopting slow step sintering schedule. It is observed that by reducing the particles size, chemical route enhances the mixing process as well as decreasing the sintering temperature to get single phase material system in compared to the polycrystalline sample prepared directly from the micron sized commercial powder. Raman spectroscopic studies confirm that the sample is in single phase without any detectable impurity. Frequency dependent dielectric properties i.e., dielectric constant (K) and dielectric loss (tanδ) of GMO ceramics sintered at 850°C for 24 hours were studied at room temperature. The sample showed high K value (˜2736) in the frequency of 100 Hz at room temperature.

SERE: single-parameter quality control and sample comparison for RNA-Seq.

PubMed

Schulze, Stefan K; Kanwar, Rahul; Gölzenleuchter, Meike; Therneau, Terry M; Beutler, Andreas S

2012-10-03

Assessing the reliability of experimental replicates (or global alterations corresponding to different experimental conditions) is a critical step in analyzing RNA-Seq data. Pearson's correlation coefficient r has been widely used in the RNA-Seq field even though its statistical characteristics may be poorly suited to the task. Here we present a single-parameter test procedure for count data, the Simple Error Ratio Estimate (SERE), that can determine whether two RNA-Seq libraries are faithful replicates or globally different. Benchmarking shows that the interpretation of SERE is unambiguous regardless of the total read count or the range of expression differences among bins (exons or genes), a score of 1 indicating faithful replication (i.e., samples are affected only by Poisson variation of individual counts), a score of 0 indicating data duplication, and scores >1 corresponding to true global differences between RNA-Seq libraries. On the contrary the interpretation of Pearson's r is generally ambiguous and highly dependent on sequencing depth and the range of expression levels inherent to the sample (difference between lowest and highest bin count). Cohen's simple Kappa results are also ambiguous and are highly dependent on the choice of bins. For quantifying global sample differences SERE performs similarly to a measure based on the negative binomial distribution yet is simpler to compute. SERE can therefore serve as a straightforward and reliable statistical procedure for the global assessment of pairs or large groups of RNA-Seq datasets by a single statistical parameter.

SERE: Single-parameter quality control and sample comparison for RNA-Seq

PubMed Central

2012-01-01

Background Assessing the reliability of experimental replicates (or global alterations corresponding to different experimental conditions) is a critical step in analyzing RNA-Seq data. Pearson’s correlation coefficient r has been widely used in the RNA-Seq field even though its statistical characteristics may be poorly suited to the task. Results Here we present a single-parameter test procedure for count data, the Simple Error Ratio Estimate (SERE), that can determine whether two RNA-Seq libraries are faithful replicates or globally different. Benchmarking shows that the interpretation of SERE is unambiguous regardless of the total read count or the range of expression differences among bins (exons or genes), a score of 1 indicating faithful replication (i.e., samples are affected only by Poisson variation of individual counts), a score of 0 indicating data duplication, and scores >1 corresponding to true global differences between RNA-Seq libraries. On the contrary the interpretation of Pearson’s r is generally ambiguous and highly dependent on sequencing depth and the range of expression levels inherent to the sample (difference between lowest and highest bin count). Cohen’s simple Kappa results are also ambiguous and are highly dependent on the choice of bins. For quantifying global sample differences SERE performs similarly to a measure based on the negative binomial distribution yet is simpler to compute. Conclusions SERE can therefore serve as a straightforward and reliable statistical procedure for the global assessment of pairs or large groups of RNA-Seq datasets by a single statistical parameter. PMID:23033915

Ion-beam assisted laser fabrication of sensing plasmonic nanostructures

PubMed Central

Kuchmizhak, Aleksandr; Gurbatov, Stanislav; Vitrik, Oleg; Kulchin, Yuri; Milichko, Valentin; Makarov, Sergey; Kudryashov, Sergey

2016-01-01

Simple high-performance, two-stage hybrid technique was developed for fabrication of different plasmonic nanostructures, including nanorods, nanorings, as well as more complex structures on glass substrates. In this technique, a thin noble-metal film on a dielectric substrate is irradiated by a single tightly focused nanosecond laser pulse and then the modified region is slowly polished by an accelerated argon ion (Ar+) beam. As a result, each nanosecond laser pulse locally modifies the initial metal film through initiation of fast melting and subsequent hydrodynamic processes, while the following Ar+-ion polishing removes the rest of the film, revealing the hidden topography features and fabricating separate plasmonic structures on the glass substrate. We demonstrate that the shape and lateral size of the resulting functional plasmonic nanostructures depend on the laser pulse energy and metal film thickness, while subsequent Ar+-ion polishing enables to vary height of the resulting nanostructures. Plasmonic properties of the fabricated nanostructures were characterized by dark-field micro-spectroscopy, Raman and photoluminescence measurements performed on single nanofeatures, as well as by supporting numerical calculations of the related electromagnetic near-fields and Purcell factors. The developed simple two-stage technique represents a new step towards direct large-scale laser-induced fabrication of highly ordered arrays of complex plasmonic nanostructures. PMID:26776569

An efficient planar accordion-shaped micromixer: from biochemical mixing to biological application

PubMed Central

Cosentino, Armando; Madadi, Hojjat; Vergara, Paola; Vecchione, Raffaele; Causa, Filippo; Netti, Paolo Antonio

2015-01-01

Micromixers are the key component that allow lab-on-a-chip and micro total analysis systems to reach the correct level of mixing for any given process. This paper proposes a novel, simple, passive micromixer design characterized by a planar accordion-shape geometry. The geometrical characteristics of the presented design were analyzed numerically in the range of 0.01 < Re < 100 based on the micromixer performance. The performance of the most efficient design was experimentally investigated by means of fluorescence microscopy for a range of low diffusion coefficients, 10−12 < D < 10−11 m2/s. The micromixer structure was fabricated in a simple single-step process using maskless lithography and soft lithography. The experimental results showed a very good agreement with the predicted numerical results. This micromixer design including a single serpentine unit (1-SERP) displayed an efficiency higher than 90% (mixing length = 6.4 mm) creating a pressure drop of about 500 Pa at Re = 0.1 and 60 kPa at Re = 10. A mixing efficiency of almost 100% was readily reached when three serpentine units were included (3-SERP). Finally, the potential diagnostic value of the presented microdevice was validated experimentally for Red Blood Cell (RBC) lysis. PMID:26658848

An efficient planar accordion-shaped micromixer: from biochemical mixing to biological application

NASA Astrophysics Data System (ADS)

Cosentino, Armando; Madadi, Hojjat; Vergara, Paola; Vecchione, Raffaele; Causa, Filippo; Netti, Paolo Antonio

2015-12-01

Micromixers are the key component that allow lab-on-a-chip and micro total analysis systems to reach the correct level of mixing for any given process. This paper proposes a novel, simple, passive micromixer design characterized by a planar accordion-shape geometry. The geometrical characteristics of the presented design were analyzed numerically in the range of 0.01 < Re < 100 based on the micromixer performance. The performance of the most efficient design was experimentally investigated by means of fluorescence microscopy for a range of low diffusion coefficients, 10-12 < D < 10-11 m2/s. The micromixer structure was fabricated in a simple single-step process using maskless lithography and soft lithography. The experimental results showed a very good agreement with the predicted numerical results. This micromixer design including a single serpentine unit (1-SERP) displayed an efficiency higher than 90% (mixing length = 6.4 mm) creating a pressure drop of about 500 Pa at Re = 0.1 and 60 kPa at Re = 10. A mixing efficiency of almost 100% was readily reached when three serpentine units were included (3-SERP). Finally, the potential diagnostic value of the presented microdevice was validated experimentally for Red Blood Cell (RBC) lysis.

An efficient planar accordion-shaped micromixer: from biochemical mixing to biological application.

PubMed

Cosentino, Armando; Madadi, Hojjat; Vergara, Paola; Vecchione, Raffaele; Causa, Filippo; Netti, Paolo Antonio

2015-12-14

Micromixers are the key component that allow lab-on-a-chip and micro total analysis systems to reach the correct level of mixing for any given process. This paper proposes a novel, simple, passive micromixer design characterized by a planar accordion-shape geometry. The geometrical characteristics of the presented design were analyzed numerically in the range of 0.01 < Re < 100 based on the micromixer performance. The performance of the most efficient design was experimentally investigated by means of fluorescence microscopy for a range of low diffusion coefficients, 10(-12) < D < 10(-11) m(2)/s. The micromixer structure was fabricated in a simple single-step process using maskless lithography and soft lithography. The experimental results showed a very good agreement with the predicted numerical results. This micromixer design including a single serpentine unit (1-SERP) displayed an efficiency higher than 90% (mixing length = 6.4 mm) creating a pressure drop of about 500 Pa at Re = 0.1 and 60 kPa at Re = 10. A mixing efficiency of almost 100% was readily reached when three serpentine units were included (3-SERP). Finally, the potential diagnostic value of the presented microdevice was validated experimentally for Red Blood Cell (RBC) lysis.

Application of a rapid and selective method for the simultaneous determination of carebastine and pseudoephedrine in human plasma by liquid chromatography-electrospray mass spectrometry for bioequivalence study in Korean subjects.

PubMed

Lee, Myung-Jae; Lee, Heon-Woo; Kang, Jong-Min; Seo, Ji-Hyung; Tak, Seong-Kun; Shim, Wangseob; Yim, Sung-Vin; Hong, Seung Jae; Lee, Kyung-Tae

2010-10-01

We describe a simple, rapid and sensitive high-performance liquid chromatography-electrospray ionization tandem mass spectrometric method that was developed for the simultaneous determination of carebastine and pseudoephedrine in human plasma using cisapride as an internal standard. Acquisition was performed in multiple-reaction monitoring mode by monitoring the transitions: m/z 500.43 > 167.09 for carebastine and m/z 166.04 > 147.88 for pseudoephedrine. The devised method involves a simple single-step liquid-liquid extraction with ethyl acetate. Chromatographic separation was performed on a C(18) reversed-phase chromatographic column at 0.2  mL/min by isocratic elution with 10  mM ammonium formate buffer-acetonitrile (30:70, v/v; adjusted to pH 3.3 with formic acid). The devised method was validated over 0.5-100  ng/mL of carebastine and 5-1000  ng/mL of pseudoephedrine with acceptable accuracy and precision, and was successfully applied to a bioequivalence study involving a single oral dose (10  mg of ebastine plus 120  mg of pseudoephedrine complex) to healthy Korean volunteers. Copyright © 2010 John Wiley & Sons, Ltd.

Simultaneous dispersive liquid-liquid microextraction derivatisation and gas chromatography mass spectrometry analysis of subcritical water extracts of sweet and sour cherry stems.

PubMed

Švarc-Gajić, Jaroslava; Clavijo, Sabrina; Suárez, Ruth; Cvetanović, Aleksandra; Cerdà, Víctor

2018-03-01

Cherry stems have been used in traditional medicine mostly for the treatment of urinary tract infections. Extraction with subcritical water, according to its selectivity, efficiency and other aspects, differs substantially from conventional extraction techniques. The complexity of plant subcritical water extracts is due to the ability of subcritical water to extract different chemical classes of different physico-chemical properties and polarities in a single run. In this paper, dispersive liquid-liquid microextraction (DLLME) with simultaneous derivatisation was optimised for the analysis of complex subcritical water extracts of cherry stems to allow simple and rapid preparation prior to gas chromatography-mass spectrometry (GC-MS). After defining optimal extracting and dispersive solvents, the optimised method was used for the identification of compounds belonging to different chemical classes in a single analytical run. The developed sample preparation protocol enabled simultaneous extraction and derivatisation, as well as convenient coupling with GC-MS analysis, reducing the analysis time and number of steps. The applied analytical protocol allowed simple and rapid chemical screening of subcritical water extracts and was used for the comparison of subcritical water extracts of sweet and sour cherry stems. Graphical abstract DLLME GC MS analysis of cherry stem extracts obtained by subcritical water.

Optimal design and critical analysis of a high resolution video plenoptic demonstrator

NASA Astrophysics Data System (ADS)

Drazic, Valter; Sacré, Jean-Jacques; Bertrand, Jérôme; Schubert, Arno; Blondé, Etienne

2011-03-01

A plenoptic camera is a natural multi-view acquisition device also capable of measuring distances by correlating a set of images acquired under different parallaxes. Its single lens and single sensor architecture have two downsides: limited resolution and depth sensitivity. In a very first step and in order to circumvent those shortcomings, we have investigated how the basic design parameters of a plenoptic camera optimize both the resolution of each view and also its depth measuring capability. In a second step, we built a prototype based on a very high resolution Red One® movie camera with an external plenoptic adapter and a relay lens. The prototype delivered 5 video views of 820x410. The main limitation in our prototype is view cross talk due to optical aberrations which reduce the depth accuracy performance. We have simulated some limiting optical aberrations and predicted its impact on the performances of the camera. In addition, we developed adjustment protocols based on a simple pattern and analyzing programs which investigate the view mapping and amount of parallax crosstalk on the sensor on a pixel basis. The results of these developments enabled us to adjust the lenslet array with a sub micrometer precision and to mark the pixels of the sensor where the views do not register properly.

Optimal design and critical analysis of a high-resolution video plenoptic demonstrator

NASA Astrophysics Data System (ADS)

Drazic, Valter; Sacré, Jean-Jacques; Schubert, Arno; Bertrand, Jérôme; Blondé, Etienne

2012-01-01

A plenoptic camera is a natural multiview acquisition device also capable of measuring distances by correlating a set of images acquired under different parallaxes. Its single lens and single sensor architecture have two downsides: limited resolution and limited depth sensitivity. As a first step and in order to circumvent those shortcomings, we investigated how the basic design parameters of a plenoptic camera optimize both the resolution of each view and its depth-measuring capability. In a second step, we built a prototype based on a very high resolution Red One® movie camera with an external plenoptic adapter and a relay lens. The prototype delivered five video views of 820 × 410. The main limitation in our prototype is view crosstalk due to optical aberrations that reduce the depth accuracy performance. We simulated some limiting optical aberrations and predicted their impact on the performance of the camera. In addition, we developed adjustment protocols based on a simple pattern and analysis of programs that investigated the view mapping and amount of parallax crosstalk on the sensor on a pixel basis. The results of these developments enabled us to adjust the lenslet array with a submicrometer precision and to mark the pixels of the sensor where the views do not register properly.

Single-step solubilization of milk samples with N,N-dimethylformamide for inductively coupled plasma-mass spectrometry analysis and classification based on their elemental composition.

PubMed

Azcarate, Silvana M; Savio, Marianela; Smichowski, Patricia; Martinez, Luis D; Camiña, José M; Gil, Raúl A

2015-10-01

A single-step procedure for trace elements analysis of milk samples is presented. Solubilization with small amounts of dymethylformamide (DMF) was assayed prior to inductively coupled plasma mass spectrometry (ICPMS) detection with a high efficiency sample introduction system. All main instrumental conditions were optimized in order to readily introduce the samples without matrix elimination. In order to assess and mitigate matrix effects in the determination of As, Cd, Co, Cu, Eu, Ga, Gd, Ge, Mn, Mo, Nb, Nd, Ni, Pb, Pr, Rb, Sm, S, Sr, Ta, Tb, V, Zn, and Zr, matrix matching calibration with (103)Rh as internal standard (IS) was performed. The obtained limits of detection were between 0.68 (Tb) and 30 (Zn) μg L(-1). For accuracy verification, certified Skim milk powder reference material (BCR 063R) was employed. The developed method was applied to trace elements analysis of commercially available milks. Principal components analysis was used to correlate the content of trace metals with the kind of milk, obtaining a classification according to adults, baby or baby fortified milks. The outcomes highlight a simple and fast approach that could be trustworthy for routine analysis, quality control and traceability of milks. Copyright © 2015 Elsevier B.V. All rights reserved.

Single-step solvothermal synthesis of mesoporous Ag-TiO2-reduced graphene oxide ternary composites with enhanced photocatalytic activity.

PubMed

Sher Shah, Md Selim Arif; Zhang, Kan; Park, A Reum; Kim, Kwang Su; Park, Nam-Gyu; Park, Jong Hyeok; Yoo, Pil J

2013-06-07

With growing interest in the photocatalytic performance of TiO2-graphene composite systems, the ternary phase of TiO2, graphene, and Ag is expected to exhibit improved photocatalytic characteristics because of the improved recombination rate of photogenerated charge carriers and potential contribution of the generation of localized surface plasmon resonance at Ag sites on a surface of the TiO2-graphene binary matrix. In this work, Ag-TiO2-reduced graphene oxide ternary nanocomposites were successfully synthesized by a simple solvothermal process. In a single-step synthetic procedure, the reduction of AgNO3 and graphene oxide and the hydrolysis of titanium tetraisopropoxide were spontaneously performed in a mixed solvent system of ethylene glycol, N,N-dimethylformamide and a stoichiometric amount of water without resorting to the use of typical reducing agents. The nanocomposites were characterized by X-ray diffraction, X-ray photoelectron spectroscopy, along with different microscopic and spectroscopic techniques, enabling us to confirm the successful reduction of AgNO3 and graphite oxide to metallic Ag and reduced graphene oxide, respectively. Due to the highly facilitated electron transport of well distributed Ag nanoparticles, the synthesized ternary nanocomposite showed enhanced photocatalytic activity for degradation of rhodamine B dye under visible light irradiation.

Single pass tangential flow filtration to debottleneck downstream processing for therapeutic antibody production.

PubMed

Dizon-Maspat, Jemelle; Bourret, Justin; D'Agostini, Anna; Li, Feng

2012-04-01

As the therapeutic monoclonal antibody (mAb) market continues to grow, optimizing production processes is becoming more critical in improving efficiencies and reducing cost-of-goods in large-scale production. With the recent trends of increasing cell culture titers from upstream process improvements, downstream capacity has become the bottleneck in many existing manufacturing facilities. Single Pass Tangential Flow Filtration (SPTFF) is an emerging technology, which is potentially useful in debottlenecking downstream capacity, especially when the pool tank size is a limiting factor. It can be integrated as part of an existing purification process, after a column chromatography step or a filtration step, without introducing a new unit operation. In this study, SPTFF technology was systematically evaluated for reducing process intermediate volumes from 2× to 10× with multiple mAbs and the impact of SPTFF on product quality, and process yield was analyzed. Finally, the potential fit into the typical 3-column industry platform antibody purification process and its implementation in a commercial scale manufacturing facility were also evaluated. Our data indicate that using SPTFF to concentrate protein pools is a simple, flexible, and robust operation, which can be implemented at various scales to improve antibody purification process capacity. Copyright © 2011 Wiley Periodicals, Inc.

Human mammary epithelial cells exhibit a bimodal correlated random walk pattern.

PubMed

Potdar, Alka A; Jeon, Junhwan; Weaver, Alissa M; Quaranta, Vito; Cummings, Peter T

2010-03-10

Organisms, at scales ranging from unicellular to mammals, have been known to exhibit foraging behavior described by random walks whose segments confirm to Lévy or exponential distributions. For the first time, we present evidence that single cells (mammary epithelial cells) that exist in multi-cellular organisms (humans) follow a bimodal correlated random walk (BCRW). Cellular tracks of MCF-10A pBabe, neuN and neuT random migration on 2-D plastic substrates, analyzed using bimodal analysis, were found to reveal the BCRW pattern. We find two types of exponentially distributed correlated flights (corresponding to what we refer to as the directional and re-orientation phases) each having its own correlation between move step-lengths within flights. The exponential distribution of flight lengths was confirmed using different analysis methods (logarithmic binning with normalization, survival frequency plots and maximum likelihood estimation). Because of the presence of non-uniform turn angle distribution of move step-lengths within a flight and two different types of flights, we propose that the epithelial random walk is a BCRW comprising of two alternating modes with varying degree of correlations, rather than a simple persistent random walk. A BCRW model rather than a simple persistent random walk correctly matches the super-diffusivity in the cell migration paths as indicated by simulations based on the BCRW model.

Cotton transformation via pollen tube pathway.

PubMed

Wang, Min; Zhang, Baohong; Wang, Qinglian

2013-01-01

Although many gene transfer methods have been employed for successfully obtaining transgenic cotton, the major constraint in cotton improvement is the limitation of genotype because the majority of transgenic methods require plant regeneration from a single transformed cell which is limited by cotton tissue culture. Comparing with other plant species, it is difficult to induce plant regeneration from cotton; currently, only a limited number of cotton cultivars can be cultured for obtaining regenerated plants. Thus, development of a simple and genotype-independent genetic transformation method is particularly important for cotton community. In this chapter, we present a simple, cost-efficient, and genotype-independent cotton transformation method-pollen tube pathway-mediated transformation. This method uses pollen tube pathway to deliver transgene into cotton embryo sacs and then insert foreign genes into cotton genome. There are three major steps for pollen tube pathway-mediated genetic transformation, which include injection of -foreign genes into pollen tube, integration of foreign genes into plant genome, and selection of transgenic plants.

Porous 3D graphene-based bulk materials with exceptional high surface area and excellent conductivity for supercapacitors

PubMed Central

Zhang, Long; Zhang, Fan; Yang, Xi; Long, Guankui; Wu, Yingpeng; Zhang, Tengfei; Leng, Kai; Huang, Yi; Ma, Yanfeng; Yu, Ao; Chen, Yongsheng

2013-01-01

Until now, few sp2 carbon materials simultaneously exhibit superior performance for specific surface area (SSA) and electrical conductivity at bulk state. Thus, it is extremely important to make such materials at bulk scale with those two outstanding properties combined together. Here, we present a simple and green but very efficient approach using two standard and simple industry steps to make such three-dimensional graphene-based porous materials at the bulk scale, with ultrahigh SSA (3523 m2/g) and excellent bulk conductivity. We conclude that these materials consist of mainly defected/wrinkled single layer graphene sheets in the dimensional size of a few nanometers, with at least some covalent bond between each other. The outstanding properties of these materials are demonstrated by their superior supercapacitor performance in ionic liquid with specific capacitance and energy density of 231 F/g and 98 Wh/kg, respectively, so far the best reported capacitance performance for all bulk carbon materials. PMID:23474952

WHEAT GERM CELL-FREE TRANSLATION, PURIFICATION, AND ASSEMBLY OF A FUNCTIONAL HUMAN STEAROYL-COA DESATURASE COMPLEX

PubMed Central

Goren, Michael A.; Fox, Brian G.

2008-01-01

A wheat germ cell-free extract was used to perform in vitro translation of human stearoyl-CoA desaturase in the presence of unilamelar liposomes, and near complete transfer of the expressed integral membrane protein into the liposome was observed. Moreover, co-translation of the desaturase along with human cytochrome b5 led to transfer of both membrane proteins into the liposomes. A simple, single step purification via centrifugation in a density gradient yielded proteoliposomes with the desaturase in high purity as judged by capillary electrophoresis. After in vitro reconstitution of the non-heme iron and heme active sites, the function of the reconstituted enzyme complex was demonstrated by conversion of stearoyl-CoA to oleoyl-CoA. This simple translation approach obviates the use of detergents or other lipids to stabilize and isolate a catalytically active integral membrane enzyme. The applicability of cell-free translation to the assembly and purification of other integral membrane protein complexes is discussed. PMID:18765284

Wheat germ cell-free translation, purification, and assembly of a functional human stearoyl-CoA desaturase complex.

PubMed

Goren, Michael A; Fox, Brian G

2008-12-01

A wheat germ cell-free extract was used to perform in vitro translation of human stearoyl-CoA desaturase in the presence of unilamelar liposomes, and near complete transfer of the expressed integral membrane protein into the liposome was observed. Moreover, co-translation of the desaturase along with human cytochrome b(5) led to transfer of both membrane proteins into the liposomes. A simple, single step purification via centrifugation in a density gradient yielded proteoliposomes with the desaturase in high purity as judged by capillary electrophoresis. After in vitro reconstitution of the non-heme iron and heme active sites, the function of the reconstituted enzyme complex was demonstrated by conversion of stearoyl-CoA to oleoyl-CoA. This simple translation approach obviates the use of detergents or other lipids to stabilize and isolate a catalytically active integral membrane enzyme. The applicability of cell-free translation to the assembly and purification of other integral membrane protein complexes is discussed.

Modified surface of titanium dioxide nanoparticles-based biosensor for DNA detection

NASA Astrophysics Data System (ADS)

Nadzirah, Sh.; Hashim, U.; Rusop, M.

2018-05-01

A new technique was used to develop a simple and selective picoammeter DNA biosensor for identification of E. coli O157:H7. This biosensor was fabricated from titanium dioxide nanoparticles that was synthesized by sol-gel method and spin-coated on silicon dioxide substrate via spinner. 3-Aminopropyl triethoxy silane (APTES) was used to modify the surface of TiO2. Simple surface modification approach has been applied; which is single dropping of APTES onto the TiO2 nanoparticles surface. Carboxyl modified probe DNA has been bind onto the surface of APTES/TiO2 without any amplifier element. Electrical signal has been used as the indicator to differentiate each step (surface modification of TiO2 and probe DNA immobilization). The I-V measurements indicate extremely low current (pico-ampere) flow through the device which is 2.8138E-10 A for pure TiO2 nanoparticles, 2.8124E-10 A after APTES modification and 3.5949E-10 A after probe DNA immobilization.

Endovascular Therapy of the Superficial Femoral Artery Via a Stand-Alone Transradial Access: A Single-Center Experience.

PubMed

Hanna, Elias B; Ababneh, Bashar A; Amin, Amit N

2018-02-01

We describe our experience in transradial recanalization of the superficial femoral artery (SFA), and we provide a stepwise approach accounting for the patient's height and optimizing the yield of currently available devices. Fifteen patients with simple SFA disease, including 4 patients with total SFA occlusions <15 cm, were selected for stand-alone transradial recanalization. A 6F, 125-cm multipurpose guiding catheter was used to cannulate the limb of interest and support device delivery. The procedure was successful in all patients and consisted of balloon angioplasty (using 0.014″, 200-cm shaft monorail balloons) in all patients, and orbital atherectomy in 6 patients. We illustrate the steps and challenges of the transradial approach, namely the limited support in complex disease and the limited reach of current equipment. In patients with simple SFA disease, transradial recanalization appears feasible and safe but currently limited to balloon angioplasty ± orbital atherectomy. Proximal SFA stenting may be feasible in patients <160 cm in height.

Quantifying prosthetic gait deviation using simple outcome measures

PubMed Central

Kark, Lauren; Odell, Ross; McIntosh, Andrew S; Simmons, Anne

2016-01-01

AIM: To develop a subset of simple outcome measures to quantify prosthetic gait deviation without needing three-dimensional gait analysis (3DGA). METHODS: Eight unilateral, transfemoral amputees and 12 unilateral, transtibial amputees were recruited. Twenty-eight able-bodied controls were recruited. All participants underwent 3DGA, the timed-up-and-go test and the six-minute walk test (6MWT). The lower-limb amputees also completed the Prosthesis Evaluation Questionnaire. Results from 3DGA were summarised using the gait deviation index (GDI), which was subsequently regressed, using stepwise regression, against the other measures. RESULTS: Step-length (SL), self-selected walking speed (SSWS) and the distance walked during the 6MWT (6MWD) were significantly correlated with GDI. The 6MWD was the strongest, single predictor of the GDI, followed by SL and SSWS. The predictive ability of the regression equations were improved following inclusion of self-report data related to mobility and prosthetic utility. CONCLUSION: This study offers a practicable alternative to quantifying kinematic deviation without the need to conduct complete 3DGA. PMID:27335814

Simple and sensitive HPLC method with fluorescence detection for the measurement of ibuprofen in rat plasma: application to a long-lasting dosage form.

PubMed

Hassan, Ahmed Sheikh; Sapin, Anne; Ubrich, Nathalie; Maincent, Philippe; Bolzan, Claire; Leroy, Pierre

2008-10-01

A simple and sensitive high-performance liquid chromatography (HPLC) assay applied to the measurement of ibuprofen in rat plasma has been developed. Two parameters have been investigated to improve ibuprofen detectability using fluorescence detection: variation of mobile phase pH and the use of beta-cyclodextrin (beta-CD). Increasing the pH value from 2.5 to 6.5 and adding 5 mM beta-CD enhanced the fluorescence signal (lambda(exc) = 224 nm; lambda(em) = 290 nm) by 2.5 and 1.3-fold, respectively, when using standards. In the case of plasma samples, only pH variation significantly lowered detection and quantification limits, down to 10 and 35 ng/mL, respectively. Full selectivity was obtained with a single step for plasma treatment, that is, protein precipitation with acidified acetonitrile. The validated method was applied to a pharmacokinetic study of ibuprofen encapsulated in microspheres and subcutaneously administered to rats.

Single-shot color fringe projection for three-dimensional shape measurement of objects with discontinuities.

PubMed

Dai, Meiling; Yang, Fujun; He, Xiaoyuan

2012-04-20

A simple but effective fringe projection profilometry is proposed to measure 3D shape by using one snapshot color sinusoidal fringe pattern. One color fringe pattern encoded with a sinusoidal fringe (as red component) and one uniform intensity pattern (as blue component) is projected by a digital video projector, and the deformed fringe pattern is recorded by a color CCD camera. The captured color fringe pattern is separated into its RGB components and division operation is applied to red and blue channels to reduce the variable reflection intensity. Shape information of the tested object is decoded by applying an arcsine algorithm on the normalized fringe pattern with subpixel resolution. In the case of fringe discontinuities caused by height steps, or spatially isolated surfaces, the separated blue component is binarized and used for correcting the phase demodulation. A simple and robust method is also introduced to compensate for nonlinear intensity response of the digital video projector. The experimental results demonstrate the validity of the proposed method.

A simplified strategy for studying the etiology of viral diseases: Apple stem grooving virus as a case study.

PubMed

Dhir, Sunny; Walia, Yashika; Zaidi, A A; Hallan, Vipin

2015-03-01

A simple method to amplify infective, complete genomes of single stranded RNA viruses by long distance PCR (LD PCR) from woody plant tissues is described in detail. The present protocol eliminates partial purification of viral particles and the amplification is achieved in three steps: (i) easy preparation of template RNA by incorporating a pre processing step before loading onto the column (ii) reverse transcription by AMV or Superscript reverse transcriptase and (iii) amplification of cDNA by LD PCR using LA or Protoscript Taq DNA polymerase. Incorporation of a preprocessing step helped to isolate consistent quality RNA from recalcitrant woody tissues such as apple, which was critical for efficient amplification of the complete genomes of Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV) and Apple chlorotic leaf spot virus (ACLSV). Complete genome of ASGV was cloned under T7 RNA polymerase promoter and was confirmed to be infectious through transcript inoculation producing symptoms similar to the wild type virus. This is the first report for the largest RNA virus genome amplified by PCR from total nucleic acid extracts of woody plant tissues. Copyright © 2014 Elsevier B.V. All rights reserved.

Hybrid Rocket Performance Prediction with Coupling Method of CFD and Thermal Conduction Calculation

NASA Astrophysics Data System (ADS)

Funami, Yuki; Shimada, Toru

The final purpose of this study is to develop a design tool for hybrid rocket engines. This tool is a computer code which will be used in order to investigate rocket performance characteristics and unsteady phenomena lasting through the burning time, such as fuel regression or combustion oscillation. When phenomena inside a combustion chamber, namely boundary layer combustion, are described, it is difficult to use rigorous models for this target. It is because calculation cost may be too expensive. Therefore simple models are required for this calculation. In this study, quasi-one-dimensional compressible Euler equations for flowfields inside a chamber and the equation for thermal conduction inside a solid fuel are numerically solved. The energy balance equation at the solid fuel surface is solved to estimate fuel regression rate. Heat feedback model is Karabeyoglu's model dependent on total mass flux. Combustion model is global single step reaction model for 4 chemical species or chemical equilibrium model for 9 chemical species. As a first step, steady-state solutions are reported.

Complex supramolecular interfacial tessellation through convergent multi-step reaction of a dissymmetric simple organic precursor

NASA Astrophysics Data System (ADS)

Zhang, Yi-Qi; Paszkiewicz, Mateusz; Du, Ping; Zhang, Liding; Lin, Tao; Chen, Zhi; Klyatskaya, Svetlana; Ruben, Mario; Seitsonen, Ari P.; Barth, Johannes V.; Klappenberger, Florian

2018-03-01

Interfacial supramolecular self-assembly represents a powerful tool for constructing regular and quasicrystalline materials. In particular, complex two-dimensional molecular tessellations, such as semi-regular Archimedean tilings with regular polygons, promise unique properties related to their nontrivial structures. However, their formation is challenging, because current methods are largely limited to the direct assembly of precursors, that is, where structure formation relies on molecular interactions without using chemical transformations. Here, we have chosen ethynyl-iodophenanthrene (which features dissymmetry in both geometry and reactivity) as a single starting precursor to generate the rare semi-regular (3.4.6.4) Archimedean tiling with long-range order on an atomically flat substrate through a multi-step reaction. Intriguingly, the individual chemical transformations converge to form a symmetric alkynyl-Ag-alkynyl complex as the new tecton in high yields. Using a combination of microscopy and X-ray spectroscopy tools, as well as computational modelling, we show that in situ generated catalytic Ag complexes mediate the tecton conversion.

Periocular dog bite injuries and responsible care.

PubMed

Burroughs, John R; Soparkar, Charles N S; Patrinely, James R; Williams, Patrick D; Holck, David E E

2002-11-01

To report patients who received dog bite injuries to the periocular area and the incidence of repeat attacks by the biting dog and its disposition. To discuss the demographics of bite attacks, occult injuries, steps to avoid medicolegal problems, and the appropriate options for animal disposition. Single-practice prospective case series of dog attacks on 18 victims. Patients were followed and questioned about any repeat attacks by the offending dog. Of 18 individuals who received dog bites to the perio-ocular area, only 2 were >12 years old. Of the 16 cases available for follow-up (4 months to 3 years; mean, 16 months), 7 of the dogs were euthanized and 1 was sent away from the family. Of the 8 remaining animals, 5 (63%) bit again. All the dogs bit family members or family friends. Although most dogs make wonderful pets, canine bites pose a significant public health concern, and some simple steps may be taken to protect against personal injury and legal fallout from repeat attacks.

Asymptotic properties of a bold random walk

NASA Astrophysics Data System (ADS)

Serva, Maurizio

2014-08-01

In a recent paper we proposed a non-Markovian random walk model with memory of the maximum distance ever reached from the starting point (home). The behavior of the walker is different from the simple symmetric random walk only when she is at this maximum distance, where, having the choice to move either farther or closer, she decides with different probabilities. If the probability of a forward step is higher than the probability of a backward step, the walker is bold and her behavior turns out to be superdiffusive; otherwise she is timorous and her behavior turns out to be subdiffusive. The scaling behavior varies continuously from subdiffusive (timorous) to superdiffusive (bold) according to a single parameter γ ∈R. We investigate here the asymptotic properties of the bold case in the nonballistic region γ ∈[0,1/2], a problem which was left partially unsolved previously. The exact results proved in this paper require new probabilistic tools which rely on the construction of appropriate martingales of the random walk and its hitting times.

Flexible fabrication of biomimetic compound eye array via two-step thermal reflow of simply pre-modeled hierarchic microstructures

NASA Astrophysics Data System (ADS)

Huang, Shengzhou; Li, Mujun; Shen, Lianguan; Qiu, Jinfeng; Zhou, Youquan

2017-06-01

A flexible fabrication method for the biomimetic compound eye (BCE) array is proposed. In this method, a triple-layer sandwich-like coating configuration was introduced, and the required hierarchic microstructures are formed with a simple single-scan exposure in maskless digital lithography. Taking advantage of the difference of glass transition point (Tg) between photoresists of each layer, the pre-formed hierarchic microstructures are in turn reflowed to the curved substrate and the BCE ommatidia in a two-step thermal reflow process. To avoid affecting the spherical substrate formed in the first thermal reflow, a non-contact strategy was proposed in the second reflow process. The measurement results were in good agreement with the designed BCE profiles. Results also showed that the fabricated BCE had good performances in optical test. The presented method is flexible, convenient, low-cost and can easily adapt to the fabrications of other optical elements with hierarchic microstructures.

One-step synthesis of solid state luminescent carbon-based silica nanohybrids for imaging of latent fingerprints

NASA Astrophysics Data System (ADS)

Li, Feng; Li, Hongren; Cui, Tianfang

2017-11-01

Fluorescent carbon-based nanomaterials(CNs) with tunable visible emission are biocompatible, environment friendly and most suitable for various biomedical applications. Despite the successes in preparing strongly fluorescent CNs, preserving the luminescence in solid materials is still challenging because of the serious emission quenching of CNs in solid state materials. In this work, fluorescent carbon and silica nanohybrids (SiCNHs) were synthesized via a simple one-step hydrothermal approach by carbonizing sodium citrate and (3-aminopropyl)triethoxysilane(APTES), and hydrolysis of tetraethyl orthosilicate(TEOS). The resultant SiCNs were characterized through X-ray diffraction (XRD), transmission electron microscopy (TEM), FT-IR, X-ray photoelectron spectroscopy (XPS), and photoluminescence (PL) spectroscopy. The SiCNs exhibited strong fluorescence in both aqueous and solid states. The luminescent solid state SiCNs power were successfully used as a fluorescent labeling material for enhanced imaging of latent fingerprints(LFPs) on single background colour and multi-coloured surfaces substrates in forensic science for individual identification.

Steps to reconcile inflationary tensor and scalar spectra

NASA Astrophysics Data System (ADS)

Miranda, Vinícius; Hu, Wayne; Adshead, Peter

2014-05-01

The recent BICEP2 B-mode polarization determination of an inflationary tensor-scalar ratio r=0.2-0.05+0.07 is in tension with simple scale-free models of inflation due to a lack of a corresponding low multipole excess in the temperature power spectrum which places a limit of r0.002<0.11 (95% C.L.) on such models. Single-field inflationary models that reconcile these two observations, even those where the tilt runs substantially, introduce a scale into the scalar power spectrum. To cancel the tensor excess, and simultaneously remove the excess already present without tensors, ideally the model should introduce this scale as a relatively sharp transition in the tensor-scalar ratio around the horizon at recombination. We consider models which generate such a step in this quantity and find that they can improve the joint fit to the temperature and polarization data by up to 2ΔlnL≈-14 without changing cosmological parameters. Precision E-mode polarization measurements should be able to test this explanation.

Optimization of Lipase-Mediated Synthesis of 1-Nonene Oxide Using Phenylacetic Acid and Hydrogen Peroxide

PubMed Central

Abdulmalek, Emilia; Arumugam, Mahashanon; Basri, Mahiran; Rahman, Mohd Basyaruddin Abdul

2012-01-01

Herein, an efficient epoxidation of 1-nonene is described. In a simple epoxidation system, commercially available Novozym 435, an immobilized Candida antarctica lipase B, and hydrogen peroxide (H2O2) were utilized to facilitate the in situ oxidation of phenylacetic acid to the corresponding peroxy acid which then reacted with 1-nonene to give 1-nonene oxide with high yield and selectivity. The aliphatic terminal alkene was epoxidised efficiently in chloroform to give an excellent yield (97%–99%) under the optimum reaction conditions, including temperature (35 °C), initial H2O2 concentration (30%), H2O2 amount (4.4 mmol), H2O2 addition rate (one step), acid amount (8.8 mmol), and stirring speed (250 rpm). Interestingly, the enzyme was stable under the single-step addition of H2O2 with a catalytic activity of 190.0 Ug−1. The entire epoxidation process was carried out within 12 h using a conventional water bath shaker. PMID:23202943

Methods and systems for detection of radionuclides

DOEpatents

Coates, Jr., John T.; DeVol, Timothy A.

2010-05-25

Disclosed are materials and systems useful in determining the existence of radionuclides in an aqueous sample. The materials provide the dual function of both extraction and scintillation to the systems. The systems can be both portable and simple to use, and as such can beneficially be utilized to determine presence and optionally concentration of radionuclide contamination in an aqueous sample at any desired location and according to a relatively simple process without the necessity of complicated sample handling techniques. The disclosed systems include a one-step process, providing simultaneous extraction and detection capability, and a two-step process, providing a first extraction step that can be carried out in a remote field location, followed by a second detection step that can be carried out in a different location.

A tailing genome walking method suitable for genomes with high local GC content.

PubMed

Liu, Taian; Fang, Yongxiang; Yao, Wenjuan; Guan, Qisai; Bai, Gang; Jing, Zhizhong

2013-10-15

The tailing genome walking strategies are simple and efficient. However, they sometimes can be restricted due to the low stringency of homo-oligomeric primers. Here we modified their conventional tailing step by adding polythymidine and polyguanine to the target single-stranded DNA (ssDNA). The tailed ssDNA was then amplified exponentially with a specific primer in the known region and a primer comprising 5' polycytosine and 3' polyadenosine. The successful application of this novel method for identifying integration sites mediated by φC31 integrase in goat genome indicates that the method is more suitable for genomes with high complexity and local GC content. Copyright © 2013 Elsevier Inc. All rights reserved.

Simple method for the generation of multiple homogeneous field volumes inside the bore of superconducting magnets

PubMed Central

Chou, Ching-Yu; Ferrage, Fabien; Aubert, Guy; Sakellariou, Dimitris

2015-01-01

Standard Magnetic Resonance magnets produce a single homogeneous field volume, where the analysis is performed. Nonetheless, several modern applications could benefit from the generation of multiple homogeneous field volumes along the axis and inside the bore of the magnet. In this communication, we propose a straightforward method using a combination of ring structures of permanent magnets in order to cancel the gradient of the stray field in a series of distinct volumes. These concepts were demonstrated numerically on an experimentally measured magnetic field profile. We discuss advantages and limitations of our method and present the key steps required for an experimental validation. PMID:26182891

Simple and sensitive method for quantification of fluorescent enzymatic mature and senescent crosslinks of collagen in bone hydrolysate using single-column high performance liquid chromatography.

PubMed

Viguet-Carrin, S; Gineyts, E; Bertholon, C; Delmas, P D

2009-01-01

A rapid high performance liquid chromatographic method was developed including an internal standard for the measurement of mature and senescent crosslinks concentration in non-demineralized bone hydrolysates. To avoid the demineralization which is a tedious step, we developed a method based on the use of a solid-phase extraction procedure to clean-up the samples. It resulted in sensitive and accurate measurements: the detection limits as low as 0.2 pmol for the pyridimium crosslinks and 0.02 pmol for the pentosidine. The inter- and intra-assay coefficients of variation were as low as 5% and 2%, respectively, for all crosslinks.

Phylogenetic Analyses: A Toolbox Expanding towards Bayesian Methods

PubMed Central

Aris-Brosou, Stéphane; Xia, Xuhua

2008-01-01

The reconstruction of phylogenies is becoming an increasingly simple activity. This is mainly due to two reasons: the democratization of computing power and the increased availability of sophisticated yet user-friendly software. This review describes some of the latest additions to the phylogenetic toolbox, along with some of their theoretical and practical limitations. It is shown that Bayesian methods are under heavy development, as they offer the possibility to solve a number of long-standing issues and to integrate several steps of the phylogenetic analyses into a single framework. Specific topics include not only phylogenetic reconstruction, but also the comparison of phylogenies, the detection of adaptive evolution, and the estimation of divergence times between species. PMID:18483574

Old and New Techniques as a Safe Hybrid Approach for Carotid Tandem Lesions.

PubMed

Barillà, David; Massara, Mafalda; Alberti, Antonino; Volpe, Alberto; Cutrupi, Andrea; Versace, Paolo; Volpe, Pietro

2016-04-01

Carotid revascularization is performed to prevent stroke. Carotid tandem lesions represent a challenge for treatment, and a hybrid approach may result effective. A high-risk 65-year-old woman presented with a "tandem lesion" of left common and internal carotid artery. She was deemed unfit for "simple" standard carotid endarterectomy (CEA). A "single-step" safe hybrid procedure was scheduled for the patient. A "Cormier" carotid vein graft bypass with a retrograde stenting was performed under local anesthesia. The "safe hybrid procedure" for tandem lesions of the common and internal carotid artery is effective and suitable in high-risk patients in a high-volume centers. Copyright © 2016 Elsevier Inc. All rights reserved.

Fabrication of the planar angular rotator using the CMOS process

NASA Astrophysics Data System (ADS)

Dai, Ching-Liang; Chang, Chien-Liu; Chen, Hung-Lin; Chang, Pei-Zen

2002-05-01

In this investigation we propose a novel planar angular rotator fabricated by the conventional complementary metal-oxide semiconductor (CMOS) process. Following the 0.6 μm single poly triple metal (SPTM) CMOS process, the device is completed by a simple maskless, post-process etching step. The rotor of the planar angular rotator rotates around its geometric center with electrostatic actuation. The proposed design adopts an intelligent mechanism including the slider-crank system to permit simultaneous motion. The CMOS planar angular rotator could be driven with driving voltages of around 40 V. The design proposed here has a shorter response time and longer life, without problems of friction and wear, compared to the more common planar angular micromotor.

Gas diffusion as a new fluidic unit operation for centrifugal microfluidic platforms.

PubMed

Ymbern, Oriol; Sández, Natàlia; Calvo-López, Antonio; Puyol, Mar; Alonso-Chamarro, Julian

2014-03-07

A centrifugal microfluidic platform prototype with an integrated membrane for gas diffusion is presented for the first time. The centrifugal platform allows multiple and parallel analysis on a single disk and integrates at least ten independent microfluidic subunits, which allow both calibration and sample determination. It is constructed with a polymeric substrate material and it is designed to perform colorimetric determinations by the use of a simple miniaturized optical detection system. The determination of three different analytes, sulfur dioxide, nitrite and carbon dioxide, is carried out as a proof of concept of a versatile microfluidic system for the determination of analytes which involve a gas diffusion separation step during the analytical procedure.

Proton irradiation of [18O]O2: production of [18F]F2 and [18F]F2 + [18F] OF2.

PubMed

Bishop, A; Satyamurthy, N; Bida, G; Hendry, G; Phelps, M; Barrio, J R

1996-04-01

The production of 18F electrophilic reagents via the 18O(p,n)18F reaction has been investigated in small-volume target bodies made of aluminum, copper, gold-plated copper and nickel, having straight or conical bore shapes. Three irradiation protocols-single-step, two-step and modified two-step-were used for the recovery of the 18F activity. The single-step irradiation protocol was tested in all the target bodies. Based on the single-step performance, aluminum targets were utilized extensively in the investigation of the two-step and modified two-step irradiation protocols. With an 11-MeV cyclotron and using the two-step irradiation protocol, > 1Ci [18F]F2 was recovered reproducibly from an aluminum target body. Probable radical mechanisms for the formation of OF2 and FONO2 (fluorine nitrate) in the single-step and modified two-step targets are proposed based on the amount of ozone generated and the nitrogen impurity present in the target gases, respectively.

Atom-Thin SnS2-xSex with Adjustable Compositions by Direct Liquid Exfoliation from Single Crystals.

PubMed

Yang, Zhanhai; Liang, Hui; Wang, Xusheng; Ma, Xinlei; Zhang, Tao; Yang, Yanlian; Xie, Liming; Chen, Dong; Long, Yujia; Chen, Jitao; Chang, Yunjie; Yan, Chunhua; Zhang, Xinxiang; Zhang, Xueji; Ge, Binghui; Ren, Zhian; Xue, Mianqi; Chen, Genfu

2016-01-26

Two-dimensional (2D) chalcogenide materials are fundamentally and technologically fascinating for their suitable band gap energy and carrier type relevant to their adjustable composition, structure, and dimensionality. Here, we demonstrate the exfoliation of single-crystal SnS2-xSex (SSS) with S/Se vacancies into an atom-thin layer by simple sonication in ethanol without additive. The introduction of vacancies at the S/Se site, the conflicting atomic radius of sulfur in selenium layers, and easy incorporation with an ethanol molecule lead to high ion accessibility; therefore, atom-thin SSS flakes can be effectively prepared by exfoliating the single crystal via sonication. The in situ pyrolysis of such materials can further adjust their compositions, representing tunable activation energy, band gap, and also tunable response to analytes of such materials. As the most basic and crucial step of the 2D material field, the successful synthesis of an uncontaminated and atom-thin sample will further push ahead the large-scale applications of 2D materials, including, but not limited to, electronics, sensing, catalysis, and energy storage fields.

Lower limb muscle moments and power during recovery from forward loss of balance in male and female single and multiple steppers.

PubMed

Carty, Christopher P; Cronin, Neil J; Lichtwark, Glen A; Mills, Peter M; Barrett, Rod S

2012-12-01

Studying recovery responses to loss of balance may help to explain why older adults are susceptible to falls. The purpose of the present study was to assess whether male and female older adults, that use a single or multiple step recovery strategy, differ in the proportion of lower limb strength used and power produced during the stepping phase of balance recovery. Eighty-four community-dwelling older adults (47 men, 37 women) participated in the study. Isometric strength of the ankle, knee and hip joint flexors and extensors was assessed using a dynamometer. Loss of balance was induced by releasing participants from a static forward lean (4 trials at each of 3 forward lean angles). Participants were instructed to recover with a single step and were subsequently classified as using a single or multiple step recovery strategy for each trial. (1) Females were weaker than males and the proportion of females that were able to recover with a single step were lower than for males at each lean magnitude. (2) Multiple compared to single steppers used a significantly higher proportion of their hip extension strength and produced less knee and ankle joint peak power during stepping, at the intermediate lean angle. Strength deficits in female compared to male participants may explain why a lower proportion of female participants were able to recover with a single step. The inability to generate sufficient power in the stepping limb appears to be a limiting factor in single step recovery from forward loss of balance. Crown Copyright © 2012. Published by Elsevier Ltd. All rights reserved.

Synthesis of BiFeO{sub 3} thin films on single-terminated Nb : SrTiO{sub 3} (111) substrates by intermittent microwave assisted hydrothermal method

DOE Office of Scientific and Technical Information (OSTI.GOV)

Velasco-Davalos, Ivan; Ambriz-Vargas, Fabian; Kolhatkar, Gitanjali

We report on a simple and fast procedure to create arrays of atomically flat terraces on single crystal SrTiO{sub 3} (111) substrates and the deposition of ferroelectric BiFeO{sub 3} thin films on such single-terminated surfaces. A microwave-assisted hydrothermal method in deionized water and ammonia solution selectively removes either (SrO{sub 3}){sup 4−} or Ti{sup 4+} layers to ensure the same chemical termination on all terraces. Measured step heights of 0.225 nm (d{sub 111}) and uniform contrast in the phase image of the terraces confirm the single termination in pure and Nb doped SrTiO{sub 3} single crystal substrates. Multiferroic BiFeO{sub 3} thinmore » films were then deposited by the same microwave assisted hydrothermal process on Nb : SrTiO{sub 3} (111) substrates. Bi(NO{sub 3}){sub 3} and Fe(NO{sub 3}){sub 3} along with KOH served as the precursors solution. Ferroelectric behavior of the BiFeO{sub 3} films on Nb : SrTiO{sub 3} (100) substrates was verified by piezoresponse force microscopy.« less

One-Pot Solvothermal in Situ Growth of 1D Single-Crystalline NiSe on Ni Foil as Efficient and Stable Transparent Conductive Oxide Free Counter Electrodes for Dye-Sensitized Solar Cells.

PubMed

Bao, Chao; Li, Faxin; Wang, Jiali; Sun, Panpan; Huang, Niu; Sun, Yihua; Fang, Liang; Wang, Lei; Sun, Xiaohua

2016-12-07

One-dimensional single-crystal nanostructural nickel selenides were successfully in situ grown on metal nickel foils by two simple one-step solvothermal methods, which formed NiSe/Ni counter electrodes (CEs) for dye-sensitized solar cells (DSSCs). The nickel foil acted as the nickel source in the reaction process, a supporting substrate, and an electron transport "speedway". Electrochemical testing indicated that the top 1D single-crystal NiSe exhibited prominent electrocatalytic activity for I 3 - reduction. Due to the metallic conductivity of Ni substrate and the outstanding electrocatalytic activity of single-crystal NiSe, the DSSC based on a NiSe/Ni CE exhibited higher fill factor (FF) and larger short-circuit current density (J sc ) than the DSSC based on Pt/FTO CE. The corresponding power conversion efficiency (6.75%) outperformed that of the latter (6.18%). Moreover, the NiSe/Ni CEs also showed excellent electrochemical stability in the I - /I 3 - redox electrolyte. These findings indicated that single-crystal NiSe in situ grown on Ni substrate was a potential candidate to replace Pt/TCO as a cheap and highly efficient counter electrode of DSSC.

An efficient fluorescent single-particle position tracking system for long-term pulsed measurements of nitrogen-vacancy centers in diamond

NASA Astrophysics Data System (ADS)

Kim, Kiho; Yun, Jiwon; Lee, Donghyuck; Kim, Dohun

2018-02-01

A simple and convenient design enables real-time three-dimensional position tracking of nitrogen-vacancy (NV) centers in diamond. The system consists entirely of commercially available components (a single-photon counter, a high-speed digital-to-analog converter, a phase-sensitive detector-based feedback device, and a piezo stage), eliminating the need for custom programming or rigorous optimization processes. With a large input range of counters and trackers combined with high sensitivity of single-photon counting, high-speed position tracking (upper bound recovery time of 0.9 s upon 250 nm of step-like positional shift) not only of bright ensembles, but also of low-photon-collection-efficiency single to few NV centers (down to 103 s-1) is possible. The tracking requires position modulation of only 10 nm, which allows simultaneous position tracking and pulsed measurements in the long term. Therefore, this tracking system enables measuring a single-spin magnetic resonance and Rabi oscillations at a very high resolution even without photon collection optimization. The system is widely applicable to various fields related to NV center quantum manipulation research such as NV optical trapping, NV tracking in fluid dynamics, and biological sensing using NV centers inside a biological cell.

Fast and scalable purification of a therapeutic full-length antibody based on process crystallization.

PubMed

Smejkal, Benjamin; Agrawal, Neeraj J; Helk, Bernhard; Schulz, Henk; Giffard, Marion; Mechelke, Matthias; Ortner, Franziska; Heckmeier, Philipp; Trout, Bernhardt L; Hekmat, Dariusch

2013-09-01

The potential of process crystallization for purification of a therapeutic monoclonal IgG1 antibody was studied. The purified antibody was crystallized in non-agitated micro-batch experiments for the first time. A direct crystallization from clarified CHO cell culture harvest was inhibited by high salt concentrations. The salt concentration of the harvest was reduced by a simple pretreatment step. The crystallization process from pretreated harvest was successfully transferred to stirred tanks and scaled-up from the mL-scale to the 1 L-scale for the first time. The crystallization yield after 24 h was 88-90%. A high purity of 98.5% was reached after a single recrystallization step. A 17-fold host cell protein reduction was achieved and DNA content was reduced below the detection limit. High biological activity of the therapeutic antibody was maintained during the crystallization, dissolving, and recrystallization steps. Crystallization was also performed with impure solutions from intermediate steps of a standard monoclonal antibody purification process. It was shown that process crystallization has a strong potential to replace Protein A chromatography. Fast dissolution of the crystals was possible. Furthermore, it was shown that crystallization can be used as a concentrating step and can replace several ultra-/diafiltration steps. Molecular modeling suggested that a negative electrostatic region with interspersed exposed hydrophobic residues on the Fv domain of this antibody is responsible for the high crystallization propensity. As a result, process crystallization, following the identification of highly crystallizable antibodies using molecular modeling tools, can be recognized as an efficient, scalable, fast, and inexpensive alternative to key steps of a standard purification process for therapeutic antibodies. Copyright © 2013 Wiley Periodicals, Inc.

Single-step electrodeposition of CIS thin films with the complexing agent triethanolamine

NASA Astrophysics Data System (ADS)

Chiu, Yu-Shuen; Hsieh, Mu-Tao; Chang, Chih-Min; Chen, Chun-Shuo; Whang, Thou-Jen

2014-04-01

Some difficulties have long been encountered by single-step electrodeposition such as the optimization of electrolyte composition, deposition potentials, deposition time, and pH values. The approach of introducing ternary components into single-step electrodeposition is rather challenging especially due to the different values of the equilibrium potential for each constituent. Complexing agents play an important role in single-step electrodeposition of CuInSe2 (CIS), since the equilibrium potential of every constituent can be brought closer to each other when complexing agents are employed. In this work, single-step electrodeposition of CIS was enhanced by adding triethanolamine (TEA) into deposition bath, the CIS thin films were improved consequently in the form of polycrystalline cauliflower structures through the examination of SEM images and XRD patterns. The optimum composition of the solution for single-step electrodeposition of CIS is found to be 5 mM CuCl2, 22 mM InCl3, and 22 mM SeO2 at pH 1.5 with 0.1 M TEA. The structures, compositions, and morphologies of as-deposited and of annealed films were investigated.

Pelvic Inflammatory Disease

MedlinePlus

... No single step can protect you from every single type of STI. Can women who have sex with women get PID? Yes. ... No single step can protect you from every single type of STI. Can women who have sex with women get PID? Yes. ...

Treatment of burns in the first 24 hours: simple and practical guide by answering 10 questions in a step-by-step form

PubMed Central

2012-01-01

Residents in training, medical students and other staff in surgical sector, emergency room (ER) and intensive care unit (ICU) or Burn Unit face a multitude of questions regarding burn care. Treatment of burns is not always straightforward. Furthermore, National and International guidelines differ from one region to another. On one hand, it is important to understand pathophysiology, classification of burns, surgical treatment, and the latest updates in burn science. On the other hand, the clinical situation for treating these cases needs clear guidelines to cover every single aspect during the treatment procedure. Thus, 10 questions have been organised and discussed in a step-by-step form in order to achieve the excellence of education and the optimal treatment of burn injuries in the first 24 hours. These 10 questions will clearly discuss referral criteria to the burn unit, primary and secondary survey, estimation of the total burned surface area (%TBSA) and the degree of burns as well as resuscitation process, routine interventions, laboratory tests, indications of Bronchoscopy and special considerations for Inhalation trauma, immediate consultations and referrals, emergency surgery and admission orders. Understanding and answering the 10 questions will not only cover the management process of Burns during the first 24 hours but also seems to be an interactive clear guide for education purpose. PMID:22583548

Ultrasonication followed by single-drop microextraction combined with GC/MS for rapid determination of organochlorine pesticides from fish.

PubMed

Shrivas, Kamlesh; Wu, Hui-Fen

2008-02-01

A novel, rapid and simple sample pretreatment technique termed ultrasonication followed by single-drop micro-extraction (U-SDME) has been developed and combined with GC/MS for the determination of organochlorine pesticides (OCPs) in fish. In the present work, the lengthy procedures generally used in the conventional methods like, Soxhlet extraction, supercritical fluid extraction, pressurized liquid extraction and microwave assisted solvent extraction for extraction of OCPs from fish tissues are minimized by the use of two simple extraction procedures. Firstly, OCPs from fish were extracted in organic solvent with ultrasonication and then subsequently preconcentrated by single-drop micro-extraction (SDME). Extraction parameters of ultrasonication and SDME were optimized in spiked sample solution in order to obtain efficient extraction of OCPs from fish tissues. The calibration curves for OCPs were found to be linear between 10-1000 ng/g with correlation of estimations in the range 0.990-0.994. The recoveries obtained in blank fish tissues were ranged from 82.1 to 95.3%. The LOD and RSD for determination of OCPs in fish were 0.5 ng/g and 9.4-10.0%, respectively. The proposed method was applied for the determination of bioconcentration factor in fish after exposure to different concentrations of OCPs in cultured water. The present method avoids the co-extraction of lipids, long extraction steps (>12 h) and large amount of organic solvent for the separation of OCPs. The main advantages of the present method are rapid, selective, sensitive and low cost for the determination of OCPs in fish.

Single conducting polymer nanowire based conductometric sensors

NASA Astrophysics Data System (ADS)

Bangar, Mangesh Ashok

The detection of toxic chemicals, gases or biological agents at very low concentrations with high sensitivity and selectivity has been subject of immense interest. Sensors employing electrical signal readout as transduction mechanism offer easy, label-free detection of target analyte in real-time. Traditional thin film sensors inherently suffered through loss of sensitivity due to current shunting across the charge depleted/added region upon analyte binding to the sensor surface, due to their large cross sectional area. This limitation was overcome by use of nanostructure such as nanowire/tube as transducer where current shunting during sensing was almost eliminated. Due to their benign chemical/electrochemical fabrication route along with excellent electrical properties and biocompatibility, conducting polymers offer cost-effective alternative over other nanostructures. Biggest obstacle in using these nanostructures is lack of easy, scalable and cost-effective way of assembling these nanostructures on prefabricated micropatterns for device fabrication. In this dissertation, three different approaches have been taken to fabricate individual or array of single conducting polymer (and metal) nanowire based devices and using polymer by itself or after functionalization with appropriate recognition molecule they have been applied for gas and biochemical detection. In the first approach electrochemical fabrication of multisegmented nanowires with middle functional Ppy segment along with ferromagnetic nickel (Ni) and end gold segments for better electrical contact was studied. This multi-layered nanowires were used along with ferromagnetic contact electrode for controlled magnetic assembly of nanowires into devices and were used for ammonia gas sensing. The second approach uses conducting polymer, polypyrrole (Ppy) nanowires using simple electrophoretic alignment and maskless electrodeposition to anchor nanowire which were further functionalized with antibodies against cancer marker protein (Cancer Antigen, CA 125) using covalent immobilization for detection of CA 125 in buffer and human blood plasma. Third approach combined electrochemical deposition of conducting polymer and assembly steps into a single step fabrication & functionalization using e-beam lithographically patterned nano-channels. Using this method array of Ppy nanowires were fabricated. Further during fabrication step, by entrapping recognition molecule (avidin) biofunctionalization was achieved. Subsequently these sensors were used for detection of biotinylated single stranded DNA.

A genotyping system capable of simultaneously analyzing >1000 single nucleotide polymorphisms in a haploid genome.

PubMed

Wang, Hui-Yun; Luo, Minjie; Tereshchenko, Irina V; Frikker, Danielle M; Cui, Xiangfeng; Li, James Y; Hu, Guohong; Chu, Yi; Azaro, Marco A; Lin, Yong; Shen, Li; Yang, Qifeng; Kambouris, Manousos E; Gao, Richeng; Shih, Weichung; Li, Honghua

2005-02-01

A high-throughput genotyping system for scoring single nucleotide polymorphisms (SNPs) has been developed. With this system, >1000 SNPs can be analyzed in a single assay, with a sensitivity that allows the use of single haploid cells as starting material. In the multiplex polymorphic sequence amplification step, instead of attaching universal sequences to the amplicons, primers that are unlikely to have nonspecific and productive interactions are used. Genotypes of SNPs are then determined by using the widely accessible microarray technology and the simple single-base extension assay. Three SNP panels, each consisting of >1000 SNPs, were incorporated into this system. The system was used to analyze 24 human genomic DNA samples. With 5 ng of human genomic DNA, the average detection rate was 98.22% when single probes were used, and 96.71% could be detected by dual probes in different directions. When single sperm cells were used, 91.88% of the SNPs were detectable, which is comparable to the level that was reached when very few genetic markers were used. By using a dual-probe assay, the average genotyping accuracy was 99.96% for 5 ng of human genomic DNA and 99.95% for single sperm. This system may be used to significantly facilitate large-scale genetic analysis even if the amount of DNA template is very limited or even highly degraded as that obtained from paraffin-embedded cancer specimens, and to make many unpractical research projects highly realistic and affordable.

Variability in Costs across Hospital Wards. A Study of Chinese Hospitals

PubMed Central

Adam, Taghreed; Evans, David B.; Ying, Bian; Murray, Christopher J. L.

2014-01-01

Introduction Analysts estimating the costs or cost-effectiveness of health interventions requiring hospitalization often cut corners because they lack data and the costs of undertaking full step-down costing studies are high. They sometimes use the costs taken from a single hospital, sometimes use simple rules of thumb for allocating total hospital costs between general inpatient care and the outpatient department, and sometimes use the average cost of an inpatient bed-day instead of a ward-specific cost. Purpose In this paper we explore for the first time the extent and the causes of variation in ward-specific costs across hospitals, using data from China. We then use the resulting model to show how ward-specific costs for hospitals outside the data set could be estimated using information on the determinants identified in the paper. Methodology Ward-specific costs estimated using step-down costing methods from 41 hospitals in 12 provinces of China were used. We used seemingly unrelated regressions to identify the determinants of variability in the ratio of the costs of specific wards to that of the outpatient department, and explain how this can be used to generate ward-specific unit costs. Findings Ward-specific unit costs varied considerably across hospitals, ranging from 1 to 24 times the unit cost in the outpatient department — average unit costs are not a good proxy for costs at specialty wards in general. The most important sources of variability were the number of staff and the level of capacity utilization. Practice Implications More careful hospital costing studies are clearly needed. In the meantime, we have shown that in China it is possible to estimate ward-specific unit costs taking into account key determinants of variability in costs across wards. This might well be a better alternative than using simple rules of thumb or using estimates from a single study. PMID:24874566

A stretchable strain sensor based on a metal nanoparticle thin film for human motion detection

NASA Astrophysics Data System (ADS)

Lee, Jaehwan; Kim, Sanghyeok; Lee, Jinjae; Yang, Daejong; Park, Byong Chon; Ryu, Seunghwa; Park, Inkyu

2014-09-01

Wearable strain sensors for human motion detection are being highlighted in various fields such as medical, entertainment and sports industry. In this paper, we propose a new type of stretchable strain sensor that can detect both tensile and compressive strains and can be fabricated by a very simple process. A silver nanoparticle (Ag NP) thin film patterned on the polydimethylsiloxane (PDMS) stamp by a single-step direct transfer process is used as the strain sensing material. The working principle is the change in the electrical resistance caused by the opening/closure of micro-cracks under mechanical deformation. The fabricated stretchable strain sensor shows highly sensitive and durable sensing performances in various tensile/compressive strains, long-term cyclic loading and relaxation tests. We demonstrate the applications of our stretchable strain sensors such as flexible pressure sensors and wearable human motion detection devices with high sensitivity, response speed and mechanical robustness.Wearable strain sensors for human motion detection are being highlighted in various fields such as medical, entertainment and sports industry. In this paper, we propose a new type of stretchable strain sensor that can detect both tensile and compressive strains and can be fabricated by a very simple process. A silver nanoparticle (Ag NP) thin film patterned on the polydimethylsiloxane (PDMS) stamp by a single-step direct transfer process is used as the strain sensing material. The working principle is the change in the electrical resistance caused by the opening/closure of micro-cracks under mechanical deformation. The fabricated stretchable strain sensor shows highly sensitive and durable sensing performances in various tensile/compressive strains, long-term cyclic loading and relaxation tests. We demonstrate the applications of our stretchable strain sensors such as flexible pressure sensors and wearable human motion detection devices with high sensitivity, response speed and mechanical robustness. Electronic supplementary information (ESI) available. See DOI: 10.1039/c4nr03295k

A successful backward step correlates with hip flexion moment of supporting limb in elderly people.

PubMed

Takeuchi, Yahiko

2018-01-01

The objective of this study was to determine the positional relationship between the center of mass (COM) and the center of pressure (COP) at the time of step landing, and to examine their relationship with the joint moments exerted by the supporting limb, with regard to factors of the successful backward step response. The study population comprised 8 community-dwelling elderly people that were observed to take successive multi steps after the landing of a backward stepping. Using a motion capture system and force plate, we measured the COM, COP and COM-COP deviation distance on landing during backward stepping. In addition, we measured the moment of the supporting limb joint during backward stepping. The multi-step data were compared with data from instances when only one step was taken (single-step). Variables that differed significantly between the single- and multi-step data were used as objective variables and the joint moments of the supporting limb were used as explanatory variables in single regression analyses. The COM-COP deviation in the anteroposterior was significantly larger in the single-step. A regression analysis with COM-COP deviation as the objective variable obtained a significant regression equation in the hip flexion moment (R2 = 0.74). The hip flexion moment of supporting limb was shown to be a significant explanatory variable in both the PS and SS phases for the relationship with COM-COP distance. This study found that to create an appropriate backward step response after an external disturbance (i.e. the ability to stop after 1 step), posterior braking of the COM by a hip flexion moment are important during the single-limbed standing phase.

Reforming Cookbook Labs

ERIC Educational Resources Information Center

Peters, Erin

2005-01-01

Deconstructing cookbook labs to require the students to be more thoughtful could break down perceived teacher barriers to inquiry learning. Simple steps that remove or disrupt the direct transfer of step-by-step procedures in cookbook labs make students think more critically about their process. Through trials in the author's middle school…

Correlation and simple linear regression.

PubMed

Eberly, Lynn E

2007-01-01

This chapter highlights important steps in using correlation and simple linear regression to address scientific questions about the association of two continuous variables with each other. These steps include estimation and inference, assessing model fit, the connection between regression and ANOVA, and study design. Examples in microbiology are used throughout. This chapter provides a framework that is helpful in understanding more complex statistical techniques, such as multiple linear regression, linear mixed effects models, logistic regression, and proportional hazards regression.

Resolving incomplete single nucleotide polymorphism tagging of HLA-DQ2.2 for coeliac disease genotyping using digital droplet PCR.

PubMed

Hardy, M Y; Ontiveros, N; Varney, M D; Tye-Din, J A

2018-04-01

A hallmark of coeliac disease (CD) is the exceptionally strong genetic association with HLA-DQ2.5, DQ8, and DQ2.2. HLA typing provides information on CD risk important to both clinicians and researchers. A method that enables simple and fast detection of all CD risk genotypes is particularly desirable for the study of large populations. Single nucleotide polymorphism (SNP)-based HLA typing can detect the CD risk genotypes by detecting a combination of six SNPs but this approach can struggle to resolve HLA-DQ2.2, seen in 4% of European CD patients, because of the low resolution of one negatively predicting SNP. We sought to optimise SNP-based HLA typing by harnessing the additional resolution of digital droplet PCR to resolve HLA-DQ2.2. Here we test this two-step approach in an unselected sample of Mexican DNA and compare its accuracy to DNA typed using traditional exon detection. The addition of digital droplet PCR for samples requiring negative prediction of HLA-DQ2.2 enabled HLA-DQ2.2 to be accurately typed. This technique is a simple addition to a SNP-based typing strategy and enables comprehensive definition of all at-risk HLA genotypes in CD in a timely and cost-effective manner. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Unexpected Reaction Pathway for butyrylcholinesterase-catalyzed inactivation of “hunger hormone” ghrelin

NASA Astrophysics Data System (ADS)

Yao, Jianzhuang; Yuan, Yaxia; Zheng, Fang; Zhan, Chang-Guo

2016-02-01

Extensive computational modeling and simulations have been carried out, in the present study, to uncover the fundamental reaction pathway for butyrylcholinesterase (BChE)-catalyzed hydrolysis of ghrelin, demonstrating that the acylation process of BChE-catalyzed hydrolysis of ghrelin follows an unprecedented single-step reaction pathway and the single-step acylation process is rate-determining. The free energy barrier (18.8 kcal/mol) calculated for the rate-determining step is reasonably close to the experimentally-derived free energy barrier (~19.4 kcal/mol), suggesting that the obtained mechanistic insights are reasonable. The single-step reaction pathway for the acylation is remarkably different from the well-known two-step acylation reaction pathway for numerous ester hydrolysis reactions catalyzed by a serine esterase. This is the first time demonstrating that a single-step reaction pathway is possible for an ester hydrolysis reaction catalyzed by a serine esterase and, therefore, one no longer can simply assume that the acylation process must follow the well-known two-step reaction pathway.

Contribution of lower limb eccentric work and different step responses to balance recovery among older adults.

PubMed

Nagano, Hanatsu; Levinger, Pazit; Downie, Calum; Hayes, Alan; Begg, Rezaul

2015-09-01

Falls during walking reflect susceptibility to balance loss and the individual's capacity to recover stability. Balance can be recovered using either one step or multiple steps but both responses are impaired with ageing. To investigate older adults' (n=15, 72.5±4.8 yrs) recovery step control a tether-release procedure was devised to induce unanticipated forward balance loss. Three-dimensional position-time data combined with foot-ground reaction forces were used to measure balance recovery. Dependent variables were; margin of stability (MoS) and available response time (ART) for spatial and temporal balance measures in the transverse and sagittal planes; lower limb joint angles and joint negative/positive work; and spatio-temporal gait parameters. Relative to multi-step responses, single-step recovery was more effective in maintaining balance, indicated by greater MoS and longer ART. MoS in the sagittal plane measure and ART in the transverse plane distinguished single step responses from multiple steps. When MoS and ART were negative (<0), balance was not secured and additional steps would be required to establish the new base of support for balance recovery. Single-step responses demonstrated greater step length and velocity and when the recovery foot landed, greater centre of mass downward velocity. Single-step strategies also showed greater ankle dorsiflexion, increased knee maximum flexion and more negative work at the ankle and knee. Collectively these findings suggest that single-step responses are more effective in forward balance recovery by directing falling momentum downward to be absorbed as lower limb eccentric work. Copyright © 2015 Elsevier B.V. All rights reserved.

Comparative analysis between clinical outcomes of primary radical resection and second completion radical resection for T2 gallbladder cancer: single-center experience.

PubMed

Cho, Seong Yeon; Park, Sang-Jae; Kim, Seong Hoon; Han, Sung-Sik; Kim, Young-Kyu; Lee, Kwang-Woong

2010-07-01

Gallbladder (GB) cancer may be discovered incidentally by histopathologic examination following simple cholecystectomy. Incidental GB cancer > or =T2 or > or =N1 needs a second radical resection. It is a matter of concern whether the prognosis may be worse in patients with T2GB cancer who undergo a second radical resection than in those who undergo primary radical resection. Between March 2001 and March 2009, 21 patients underwent a one-step operation (OSO group), and 17 patients underwent a two-step operation (TSO group) for T2GB cancer. We compared clinicopathologic factors and survival between patients in the OSO group (n = 9) and those in the TSO group (n = 9) with T2N0M0 GB cancer and between patients in the OSO group (n = 12) and those in the TSO group (n = 8) with T2N1M0 GB cancer. Except for patient age, clinicopathologic factors as well as disease-free survival were not significantly different between the OSO group and the TSO group in the aforementioned cancer stages. Patient age was significantly higher in the OSO group than in the TSO group. Second completion radical resection following initial simple cholecystectomy (TSO) provided a survival benefit similar to that of primary radical surgery (OSO) for patients with both T2N0M0 and T2N1M0 GB cancers in our study.

Convolutional neural networks for vibrational spectroscopic data analysis.

PubMed

Acquarelli, Jacopo; van Laarhoven, Twan; Gerretzen, Jan; Tran, Thanh N; Buydens, Lutgarde M C; Marchiori, Elena

2017-02-15

In this work we show that convolutional neural networks (CNNs) can be efficiently used to classify vibrational spectroscopic data and identify important spectral regions. CNNs are the current state-of-the-art in image classification and speech recognition and can learn interpretable representations of the data. These characteristics make CNNs a good candidate for reducing the need for preprocessing and for highlighting important spectral regions, both of which are crucial steps in the analysis of vibrational spectroscopic data. Chemometric analysis of vibrational spectroscopic data often relies on preprocessing methods involving baseline correction, scatter correction and noise removal, which are applied to the spectra prior to model building. Preprocessing is a critical step because even in simple problems using 'reasonable' preprocessing methods may decrease the performance of the final model. We develop a new CNN based method and provide an accompanying publicly available software. It is based on a simple CNN architecture with a single convolutional layer (a so-called shallow CNN). Our method outperforms standard classification algorithms used in chemometrics (e.g. PLS) in terms of accuracy when applied to non-preprocessed test data (86% average accuracy compared to the 62% achieved by PLS), and it achieves better performance even on preprocessed test data (96% average accuracy compared to the 89% achieved by PLS). For interpretability purposes, our method includes a procedure for finding important spectral regions, thereby facilitating qualitative interpretation of results. Copyright © 2016 Elsevier B.V. All rights reserved.

Comparing an annual and daily time-step model for predicting field-scale phosphorus loss

USDA-ARS?s Scientific Manuscript database

Numerous models exist for describing phosphorus (P) losses from agricultural fields. The complexity of these models varies considerably ranging from simple empirically-based annual time-step models to more complex process-based daily time step models. While better accuracy is often assumed with more...

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

EPA Science Inventory

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 ...

The parallel-antiparallel signal difference in double-wave-vector diffusion-weighted MR at short mixing times: A phase evolution perspective

NASA Astrophysics Data System (ADS)

Finsterbusch, Jürgen

2011-01-01

Experiments with two diffusion weightings applied in direct succession in a single acquisition, so-called double- or two-wave-vector diffusion-weighting (DWV) experiments at short mixing times, have been shown to be a promising tool to estimate cell or compartment sizes, e.g. in living tissue. The basic theory for such experiments predicts that the signal decays for parallel and antiparallel wave vector orientations differ by a factor of three for small wave vectors. This seems to be surprising because in standard, single-wave-vector experiments the polarity of the diffusion weighting has no influence on the signal attenuation. Thus, the question how this difference can be understood more pictorially is often raised. In this rather educational manuscript, the phase evolution during a DWV experiment for simple geometries, e.g. diffusion between parallel, impermeable planes oriented perpendicular to the wave vectors, is considered step-by-step and demonstrates how the signal difference develops. Considering the populations of the phase distributions obtained, the factor of three between the signal decays which is predicted by the theory can be reproduced. Furthermore, the intermediate signal decay for orthogonal wave vector orientations can be derived when investigating diffusion in a box. Thus, the presented “phase gymnastics” approach may help to understand the signal modulation observed in DWV experiments at short mixing times.

Simple platform for chronic imaging of hippocampal activity during spontaneous behaviour in an awake mouse

PubMed Central

Villette, Vincent; Levesque, Mathieu; Miled, Amine; Gosselin, Benoit; Topolnik, Lisa

2017-01-01

Chronic electrophysiological recordings of neuronal activity combined with two-photon Ca2+ imaging give access to high resolution and cellular specificity. In addition, awake drug-free experimentation is required for investigating the physiological mechanisms that operate in the brain. Here, we developed a simple head fixation platform, which allows simultaneous chronic imaging and electrophysiological recordings to be obtained from the hippocampus of awake mice. We performed quantitative analyses of spontaneous animal behaviour, the associated network states and the cellular activities in the dorsal hippocampus as well as estimated the brain stability limits to image dendritic processes and individual axonal boutons. Ca2+ imaging recordings revealed a relatively stereotyped hippocampal activity despite a high inter-animal and inter-day variability in the mouse behavior. In addition to quiet state and locomotion behavioural patterns, the platform allowed the reliable detection of walking steps and fine speed variations. The brain motion during locomotion was limited to ~1.8 μm, thus allowing for imaging of small sub-cellular structures to be performed in parallel with recordings of network and behavioural states. This simple device extends the drug-free experimentation in vivo, enabling high-stability optophysiological experiments with single-bouton resolution in the mouse awake brain. PMID:28240275

A Single-Step Enrichment Medium for Nonchromogenic Isolation of Healthy and Cold-Injured Salmonella spp. from Fresh Vegetables.

PubMed

Kim, Hong-Seok; Choi, Dasom; Kang, Il-Byeong; Kim, Dong-Hyeon; Yim, Jin-Hyeok; Kim, Young-Ji; Chon, Jung-Whan; Oh, Deog-Hwan; Seo, Kun-Ho

2017-02-01

Culture-based detection of nontyphoidal Salmonella spp. in foods requires at least four working days; therefore, new detection methods that shorten the test time are needed. In this study, we developed a novel single-step Salmonella enrichment broth, SSE-1, and compared its detection capability with that of commercial single-step ONE broth-Salmonella (OBS) medium and a conventional two-step enrichment method using buffered peptone water and Rappaport-Vassiliadis soy broth (BPW-RVS). Minimally processed lettuce samples were artificially inoculated with low levels of healthy and cold-injured Salmonella Enteritidis (10 0 or 10 1 colony-forming unit/25 g), incubated in OBS, BPW-RVS, and SSE-1 broths, and streaked on xylose lysine deoxycholate (XLD) agar. Salmonella recoverability was significantly higher in BPW-RVS (79.2%) and SSE-1 (83.3%) compared to OBS (39.3%) (p < 0.05). Our data suggest that the SSE-1 single-step enrichment broth could completely replace two-step enrichment with reduced enrichment time from 48 to 24 h, performing better than commercial single-step enrichment medium in the conventional nonchromogenic Salmonella detection, thus saving time, labor, and cost.

A hierarchical nanostructure consisting of amorphous MnO 2, Mn 3O 4 nanocrystallites, and single-crystalline MnOOH nanowires for supercapacitors

NASA Astrophysics Data System (ADS)

Hu, Chi-Chang; Hung, Ching-Yun; Chang, Kuo-Hsin; Yang, Yi-Lin

In this communication, a porous hierarchical nanostructure consisting of amorphous MnO 2 (a-MnO 2), Mn 3O 4 nanocrystals, and single-crystalline MnOOH nanowires is designed for the supercapacitor application, which is prepared by a simple two-step electrochemical deposition process. Because of the gradual co-transformation of Mn 3O 4 nanocrystals and a-MnO 2 nanorods into an amorphous manganese oxide, the cycle stability of a-MnO 2 is obviously enhanced by adding Mn 3O 4. This unique ternary oxide nanocomposite with 100-cycle CV activation exhibits excellent capacitive performances, i.e., excellent reversibility, high specific capacitances (470 F g -1 in CaCl 2), high power property, and outstanding cycle stability. The highly porous microstructures of this composite before and after the 10,000-cycle CV test are examined by means of scanning electron microscopy (SEM) and transmission electron microscopy (TEM).

A high performance transparent resistive switching memory made from ZrO2/AlON bilayer structure

NASA Astrophysics Data System (ADS)

Tsai, Tsung-Ling; Chang, Hsiang-Yu; Lou, Jesse Jen-Chung; Tseng, Tseung-Yuen

2016-04-01

In this study, the switching properties of an indium tin oxide (ITO)/zirconium oxide (ZrO2)/ITO single layer device and those of a device with an aluminum oxynitride (AlON) layer were investigated. The devices with highly transparent characteristics were fabricated. Compared with the ITO/ZrO2/ITO single layer device, the ITO/ZrO2/AlON/ITO bilayer device exhibited a larger ON/OFF ratio, higher endurance performance, and superior retention properties by using a simple two-step forming process. These substantial improvements in the resistive switching properties were attributed to the minimized influence of oxygen migration through the ITO top electrode (TE), which can be realized by forming an asymmetrical conductive filament with the weakest part at the ZrO2/AlON interface. Therefore, in the ITO/ZrO2/AlON/ITO bilayer device, the regions where conductive filament formation and rupture occur can be effectively moved from the TE interface to the interior of the device.

Domain walls in single-chain magnets

NASA Astrophysics Data System (ADS)

Pianet, Vivien; Urdampilleta, Matias; Colin, Thierry; Clérac, Rodolphe; Coulon, Claude

2017-12-01

The topology and creation energy of domain walls in different magnetic chains (called Single-Chain Magnets or SCMs) are discussed. As these domain walls, that can be seen as "defects", are known to control both static and dynamic properties of these one-dimensional systems, their study and understanding are necessary first steps before a deeper discussion of the SCM properties at finite temperature. The starting point of the paper is the simple regular ferromagnetic chain for which the characteristics of the domain walls are well known. Then two cases will be discussed (i) the "mixed chains" in which isotropic and anisotropic classical spins alternate, and (ii) the so-called "canted chains" where two different easy axis directions are present. In particular, we show that "strictly narrow" domain walls no longer exist in these more complex cases, while a cascade of phase transitions is found for canted chains as the canting angle approaches 45∘. The consequence for thermodynamic properties is briefly discussed in the last part of the paper.

MIG-seq: an effective PCR-based method for genome-wide single-nucleotide polymorphism genotyping using the next-generation sequencing platform

PubMed Central

Suyama, Yoshihisa; Matsuki, Yu

2015-01-01

Restriction-enzyme (RE)-based next-generation sequencing methods have revolutionized marker-assisted genetic studies; however, the use of REs has limited their widespread adoption, especially in field samples with low-quality DNA and/or small quantities of DNA. Here, we developed a PCR-based procedure to construct reduced representation libraries without RE digestion steps, representing de novo single-nucleotide polymorphism discovery, and its genotyping using next-generation sequencing. Using multiplexed inter-simple sequence repeat (ISSR) primers, thousands of genome-wide regions were amplified effectively from a wide variety of genomes, without prior genetic information. We demonstrated: 1) Mendelian gametic segregation of the discovered variants; 2) reproducibility of genotyping by checking its applicability for individual identification; and 3) applicability in a wide variety of species by checking standard population genetic analysis. This approach, called multiplexed ISSR genotyping by sequencing, should be applicable to many marker-assisted genetic studies with a wide range of DNA qualities and quantities. PMID:26593239

Simple processing of back-contacted silicon heterojunction solar cells using selective-area crystalline growth

NASA Astrophysics Data System (ADS)

Tomasi, Andrea; Paviet-Salomon, Bertrand; Jeangros, Quentin; Haschke, Jan; Christmann, Gabriel; Barraud, Loris; Descoeudres, Antoine; Seif, Johannes Peter; Nicolay, Sylvain; Despeisse, Matthieu; de Wolf, Stefaan; Ballif, Christophe

2017-04-01

For crystalline-silicon solar cells, voltages close to the theoretical limit are nowadays readily achievable when using passivating contacts. Conversely, maximal current generation requires the integration of the electron and hole contacts at the back of the solar cell to liberate its front from any shadowing loss. Recently, the world-record efficiency for crystalline-silicon single-junction solar cells was achieved by merging these two approaches in a single device; however, the complexity of fabricating this class of devices raises concerns about their commercial potential. Here we show a contacting method that substantially simplifies the architecture and fabrication of back-contacted silicon solar cells. We exploit the surface-dependent growth of silicon thin films, deposited by plasma processes, to eliminate the patterning of one of the doped carrier-collecting layers. Then, using only one alignment step for electrode definition, we fabricate a proof-of-concept 9-cm2 tunnel-interdigitated back-contact solar cell with a certified conversion efficiency >22.5%.

Localized transfection on arrays of magnetic beads coated with PCR products.

PubMed

Isalan, Mark; Santori, Maria Isabel; Gonzalez, Cayetano; Serrano, Luis

2005-02-01

High-throughput gene analysis would benefit from new approaches for delivering DNA or RNA into cells. Here we describe a simple system that allows any molecular biology laboratory to carry out multiple, parallel cell transfections on microscope coverslip arrays. By using magnetically defined positions and PCR product-coated paramagnetic beads, we achieved transfection in a variety of cell lines. Beads may be added to the cells at any time, allowing both spatial and temporal control of transfection. Because the beads may be coated with more than one gene construct, the method can be used to achieve cotransfection within single cells. Furthermore, PCR-generated mutants may be conveniently screened, bypassing cloning and plasmid purification steps. We illustrated the applicability of the method by screening combinatorial peptide libraries, fused to GFP, to identify previously unknown cellular localization motifs. In this way, we identified several localizing peptides, including structured localization signals based around the scaffold of a single C2H2 zinc finger.

Selective emitter solar cell formation by NH3 plasma nitridation and single diffusion

NASA Astrophysics Data System (ADS)

Wu, Yung-Hsien; Chen, Lun-Lun; Wu, Jia-Rong; Wu, Min-Lin

2010-01-01

A new and simple process for fabricating a selective emitter solar cell has been proposed. Lightly and heavily doped emitters could be concurrently formed after a single POCl3 diffusion step through the selective formation of SiNx, which serves as the diffusion barrier and can be grown by NH3 plasma nitridation of the Si surface. The desired phosphorus depth profile for the lightly and heavily doped region verifies the eligibility of this process. From the electrical characterization, the selective emitter solar cell fabricated by this process manifests a higher absolute conversion efficiency than a conventional one by 0.5%. It is the enhanced response to the short wavelength light and the reduced surface recombination that causes the considerable improvement in conversion efficiency which is beneficial to further hold the competitive advantage for solar cell manufacturers. Most importantly, the proposed process can be fully integrated into the conventional solar cell process in a mass-production laboratory.

Blocking monocyte transmigration in in vitro system by an anti-CD99 human antibody in single chain fragment variable (scFv) format. Efficient large scale purification of biological active scFv from inclusion bodies in E. coli expression system

PubMed Central

Moricoli, Diego; Muller, William A.; Carbonella, Damiano Cosimo; Balducci, Maria Cristina; Dominici, Sabrina; Fiori, Valentina; Watson, Richard; Weber, Evan; Cianfriglia, Maurizio; Scotlandi, Katia; Magnani, Mauro

2015-01-01

Migration of leukocytes into a site of inflammation involves several steps mediated by various families of adhesion molecules. CD99 play a significant role in transendothelial migration (TEM) of leukocytes. Inhibition of TEM by specific monoclonal antibody (mAb) can provide a potent therapeutic approach to treating inflammatory conditions. However, the therapeutic utilization of whole IgG can lead to an inappropriate activation of Fc receptor-expressing cells inducing serious adverse side effects due to cytokine release. In this regard, specific recombinant antibody in single chain variable fragments (scFvs) originated by phage library may offer a solution by affecting TEM function in a safe clinical context. However, this consideration requires large scale production of functional scFv antibodies under GMP conditions and hence, the absence of toxic reagents utilized for the solubilization and refolding steps of inclusion bodies that may discourage industrial application of these antibody fragments. In order to apply the scFv anti-CD99 named C7A in a clinical setting we herein describe an efficient and large scale production of the antibody fragments expressed in E.coli as insoluble protein avoiding gel filtration chromatography approach, and laborious refolding step pre- and post-purification. Using differential salt elution which is a simple, reproducible and effective procedure we are able to separate scFv in monomer format from aggregates. The purified scFv antibody C7A exhibits inhibitory activity comparable to an antagonistic conventional mAb, thus providing an excellent agent for blocking CD99 signalling. Thanks to the original purification protocol that can be extended to other scFvs that are expressed as inclusion bodies in bacterial systems, the scFv anti-CD99 C7A herein described represents the first step towards the construction of new antibody therapeutic. PMID:24798881

Implementing successful strategic plans: a simple formula.

PubMed

Blondeau, Whitney; Blondeau, Benoit

2015-01-01

Strategic planning is a process. One way to think of strategic planning is to envision its development and design as a framework that will help your hospital navigate through internal and external changing environments over time. Although the process of strategic planning can feel daunting, following a simple formula involving five steps using the mnemonic B.E.G.I.N. (Begin, Evaluate, Goals & Objectives, Integration, and Next steps) will help the planning process feel more manageable, and lead you to greater success.

Improving the gate fidelity of capacitively coupled spin qubits

NASA Astrophysics Data System (ADS)

Wang, Xin; Barnes, Edwin

2015-03-01

Precise execution of quantum gates acting on two or multiple qubits is essential to quantum computation. For semiconductor spin qubits coupled via capacitive interaction, the best fidelity for a two-qubit gate demonstrated so far is around 70%, insufficient for fault-tolerant quantum computation. In this talk we present control protocols that may substantially improve the robustness of two-qubit gates against both nuclear noise and charge noise. Our pulse sequences incorporate simultaneous dynamical decoupling protocols and are simple enough for immediate experimental realization. Together with existing control protocols for single-qubit gates, our results constitute an important step toward scalable quantum computation using spin qubits. This work is done in collaboration with Sankar Das Sarma and supported by LPS-NSA-CMTC and IARPA-MQCO.

Unconstrained Capacities of Quantum Key Distribution and Entanglement Distillation for Pure-Loss Bosonic Broadcast Channels.

PubMed

Takeoka, Masahiro; Seshadreesan, Kaushik P; Wilde, Mark M

2017-10-13

We consider quantum key distribution (QKD) and entanglement distribution using a single-sender multiple-receiver pure-loss bosonic broadcast channel. We determine the unconstrained capacity region for the distillation of bipartite entanglement and secret key between the sender and each receiver, whenever they are allowed arbitrary public classical communication. A practical implication of our result is that the capacity region demonstrated drastically improves upon rates achievable using a naive time-sharing strategy, which has been employed in previously demonstrated network QKD systems. We show a simple example of a broadcast QKD protocol overcoming the limit of the point-to-point strategy. Our result is thus an important step toward opening a new framework of network channel-based quantum communication technology.

Repeated Solid-state Dewetting of Thin Gold Films for Nanogap-rich Plasmonic Nanoislands.

PubMed

Kang, Minhee; Park, Sang-Gil; Jeong, Ki-Hun

2015-10-15

This work reports a facile wafer-level fabrication for nanogap-rich gold nanoislands for highly sensitive surface enhanced Raman scattering (SERS) by repeating solid-state thermal dewetting of thin gold film. The method provides enlarged gold nanoislands with small gap spacing, which increase the number of electromagnetic hotspots and thus enhance the extinction intensity as well as the tunability for plasmon resonance wavelength. The plasmonic nanoislands from repeated dewetting substantially increase SERS enhancement factor over one order-of-magnitude higher than those from a single-step dewetting process and they allow ultrasensitive SERS detection of a neurotransmitter with extremely low Raman activity. This simple method provides many opportunities for engineering plasmonics for ultrasensitive detection and highly efficient photon collection.

Repeated Solid-state Dewetting of Thin Gold Films for Nanogap-rich Plasmonic Nanoislands

PubMed Central

Kang, Minhee; Park, Sang-Gil; Jeong, Ki-Hun

2015-01-01

This work reports a facile wafer-level fabrication for nanogap-rich gold nanoislands for highly sensitive surface enhanced Raman scattering (SERS) by repeating solid-state thermal dewetting of thin gold film. The method provides enlarged gold nanoislands with small gap spacing, which increase the number of electromagnetic hotspots and thus enhance the extinction intensity as well as the tunability for plasmon resonance wavelength. The plasmonic nanoislands from repeated dewetting substantially increase SERS enhancement factor over one order-of-magnitude higher than those from a single-step dewetting process and they allow ultrasensitive SERS detection of a neurotransmitter with extremely low Raman activity. This simple method provides many opportunities for engineering plasmonics for ultrasensitive detection and highly efficient photon collection. PMID:26469768

Comparison of Several Dissipation Algorithms for Central Difference Schemes

NASA Technical Reports Server (NTRS)

Swanson, R. C.; Radespiel, R.; Turkel, E.

1997-01-01

Several algorithms for introducing artificial dissipation into a central difference approximation to the Euler and Navier Stokes equations are considered. The focus of the paper is on the convective upwind and split pressure (CUSP) scheme, which is designed to support single interior point discrete shock waves. This scheme is analyzed and compared in detail with scalar and matrix dissipation (MATD) schemes. Resolution capability is determined by solving subsonic, transonic, and hypersonic flow problems. A finite-volume discretization and a multistage time-stepping scheme with multigrid are used to compute solutions to the flow equations. Numerical results are also compared with either theoretical solutions or experimental data. For transonic airfoil flows the best accuracy on coarse meshes for aerodynamic coefficients is obtained with a simple MATD scheme.

A miniaturized assay for measuring small molecule phosphorylation in the presence of complex matrices.

PubMed

Spry, Christina; Saliba, Kevin J; Strauss, Erick

2014-04-15

We describe here a simple, miniaturized radiation-based phosphorylation assay that can be used to monitor phosphorylation of a diverse range of small molecule substrates in the presence of purified and crude enzyme preparations. Ba(OH)2 and ZnSO4 are used to terminate phosphoryl transfer and to precipitate selectively the phosphorylated reaction product in a single step; non-phosphorylated substrate is removed by filtration prior to quantification. The key advantages over alternative radiation-based assays are that: (i) high-energy/short-lived radioactive emitters are not required; (ii) high-quality data can be obtained without the need for high radioactivity concentrations; and (iii) the assay is compatible with high-throughput applications. Copyright © 2013 Elsevier Inc. All rights reserved.

Stereocontrolled semi-syntheses of deguelin and tephrosin† †Electronic supplementary information (ESI) available: 1H and 13C NMR spectra. See DOI: 10.1039/c6ob02659a Click here for additional data file.

PubMed Central

Russell, David A.; Freudenreich, Julien J.; Ciardiello, Joe J.; Sore, Hannah F.

2017-01-01

We describe stereocontrolled semi-syntheses of deguelin and tephrosin, anti-cancer rotenoids isolated from Tephrosia vogelii. Firstly, we present a new two-step transformation of rotenone into rot-2′-enonic acid via a zinc-mediated ring opening of rotenone hydrobromide. Secondly, following conversion of rot-2′-enonic acid into deguelin, a chromium-mediated hydroxylation provides tephrosin as a single diastereoisomer. An Étard-like reaction mechanism is proposed to account for the stereochemical outcome. Our syntheses of deguelin and tephrosin are operationally simple, scalable and high yielding, offering considerable advantages over previous methods. PMID:28134391

A mechanical jig for measuring ankle supination and pronation torque in vitro and in vivo.

PubMed

Fong, Daniel Tik-Pui; Chung, Mandy Man-Ling; Chan, Yue-Yan; Chan, Kai-Ming

2012-07-01

This study presents the design of a mechanical jig for evaluating the ankle joint torque on both cadaver and human ankles. Previous study showed that ankle sprain motion was a combination of plantarflexion and inversion. The device allows measurement of ankle supination and pronation torque with one simple axis in a single step motion. More importantly, the ankle orientation allows rotation starting from an anatomical position. Six cadaveric specimens and six human subjects were tested with simulated and voluntary rotation respectively. The presented mechanical jig makes possible the determination of supination torque for studying ankle sprain injury and the estimation of pronation torque for examining peroneal muscle response. Copyright © 2012 IPEM. Published by Elsevier Ltd. All rights reserved.

Applications of laser-induced periodic surface structures (LIPSS)

NASA Astrophysics Data System (ADS)

Bonse, Jörn; Kirner, Sabrina V.; Höhm, Sandra; Epperlein, Nadja; Spaltmann, Dirk; Rosenfeld, Arkadi; Krüger, Jörg

2017-02-01

Laser-induced periodic surface structures (LIPSS, ripples) are a universal phenomenon that can be observed on almost any material after the irradiation by linearly polarized laser beams, particularly when using ultrashort laser pulses with durations in the picosecond to femtosecond range. During the past few years significantly increasing research activities have been reported in the field of LIPSS, since their generation in a single-step process provides a simple way of nanostructuring and surface functionalization towards the control of optical, mechanical or chemical properties. In this contribution current applications of LIPSS are reviewed, including the colorization of technical surfaces, the control of surface wetting, the tailoring of surface colonization by bacterial biofilms, and the improvement of the tribological performance of nanostructured metal surfaces.

Single Visit Feeding Appliance for 1-day-old Neonate with Cleft Palate Using Safe Dental Putty-Gauze Hybrid Impression Technique for Maxillary Impression.

PubMed

Rathee, Manu

2015-01-01

Cleft lip and palate is one of the most common craniofacial anomalies of humans. Intraoral impression making is the first clinical step in the fabrication of feeding appliance for infants with oro-nasal communication. It is difficult to control the flow of the impression material in the cleft area and undercuts in a child patient. This clinical report presents a simple and safe impression technique for maxillary impression making in neonates and infants with cleft palate. A gauze piece was used to confine the impression material during functional movements of sucking while impression making in an awake child to avoid the risk of aspiration or swallowing.

Does my step look big in this? A visual illusion leads to safer stepping behaviour.

PubMed

Elliott, David B; Vale, Anna; Whitaker, David; Buckley, John G

2009-01-01

Tripping is a common factor in falls and a typical safety strategy to avoid tripping on steps or stairs is to increase foot clearance over the step edge. In the present study we asked whether the perceived height of a step could be increased using a visual illusion and whether this would lead to the adoption of a safer stepping strategy, in terms of greater foot clearance over the step edge. The study also addressed the controversial question of whether motor actions are dissociated from visual perception. 21 young, healthy subjects perceived the step to be higher in a configuration of the horizontal-vertical illusion compared to a reverse configuration (p = 0.01). During a simple stepping task, maximum toe elevation changed by an amount corresponding to the size of the visual illusion (p<0.001). Linear regression analyses showed highly significant associations between perceived step height and maximum toe elevation for all conditions. The perceived height of a step can be manipulated using a simple visual illusion, leading to the adoption of a safer stepping strategy in terms of greater foot clearance over a step edge. In addition, the strong link found between perception of a visual illusion and visuomotor action provides additional support to the view that the original, controversial proposal by Goodale and Milner (1992) of two separate and distinct visual streams for perception and visuomotor action should be re-evaluated.

The stepping behavior analysis of pedestrians from different age groups via a single-file experiment

NASA Astrophysics Data System (ADS)

Cao, Shuchao; Zhang, Jun; Song, Weiguo; Shi, Chang'an; Zhang, Ruifang

2018-03-01

The stepping behavior of pedestrians with different age compositions in single-file experiment is investigated in this paper. The relation between step length, step width and stepping time are analyzed by using the step measurement method based on the calculation of curvature of the trajectory. The relations of velocity-step width, velocity-step length and velocity-stepping time for different age groups are discussed and compared with previous studies. Finally effects of pedestrian gender and height on stepping laws and fundamental diagrams are analyzed. The study is helpful for understanding pedestrian dynamics of movement. Meanwhile, it offers experimental data to develop a microscopic model of pedestrian movement by considering stepping behavior.

Single-step preparation of selected biological fluids for the high performance liquid chromatographic analysis of fat-soluble vitamins and antioxidants.

PubMed

Lazzarino, Giacomo; Longo, Salvatore; Amorini, Angela Maria; Di Pietro, Valentina; D'Urso, Serafina; Lazzarino, Giuseppe; Belli, Antonio; Tavazzi, Barbara

2017-12-08

Fat-soluble vitamins and antioxidants are of relevance in health and disease. Current methods to extract these compounds from biological fluids mainly need use of multi-steps and multi organic solvents. They are time-consuming and difficult to apply to treat simultaneously large sample number. We here describe a single-step, one solvent extraction of fat-soluble vitamins and antioxidants from biological fluids, and the chromatographic separation of all-trans-retinoic acid, 25-hydroxycholecalciferol, all-trans-retinol, astaxanthin, lutein, zeaxanthin, trans-β-apo-8'-carotenal, γ-tocopherol, β-cryptoxanthin, α-tocopherol, phylloquinone, lycopene, α-carotene, β-carotene and coenzyme Q 10 . Extraction is obtained by adding one volume of biological fluid to two acetonitrile volumes, vortexing for 60s and incubating for 60min at 37°C under agitation. HPLC separation occurs in 30min using Hypersil C18, 100×4.6mm, 5μm particle size column, gradient from 70% methanol+30% H 2 O to 100% acetonitrile, flow rate of 1.0ml/min and 37°C column temperature. Compounds are revealed using highly sensitive UV-VIS diode array detector. The HPLC method suitability was assessed in terms of sensitivity, reproducibility and recovery. Using the present extraction and chromatographic conditions we obtained values of the fat-soluble vitamins and antioxidants in serum from 50 healthy controls similar to those found in literature. Additionally, the profile of these compounds was also measured in seminal plasma from 20 healthy fertile donors. Results indicate that this simple, rapid and low cost sample processing is suitable to extract fat-soluble vitamins and antioxidants from biological fluids and can be applied in clinical and nutritional studies. Copyright © 2017 Elsevier B.V. All rights reserved.

Real-time fluorescence ligase chain reaction for sensitive detection of single nucleotide polymorphism based on fluorescence resonance energy transfer.

PubMed

Sun, Yueying; Lu, Xiaohui; Su, Fengxia; Wang, Limei; Liu, Chenghui; Duan, Xinrui; Li, Zhengping

2015-12-15

Most of practical methods for detection of single nucleotide polymorphism (SNP) need at least two steps: amplification (usually by PCR) and detection of SNP by using the amplification products. Ligase chain reaction (LCR) can integrate the amplification and allele discrimination in one step. However, the detection of LCR products still remains a great challenge for highly sensitive and quantitative SNP detection. Herein, a simple but robust strategy for real-time fluorescence LCR has been developed for highly sensitive and quantitative SNP detection. A pair of LCR probes are firstly labeled with a fluorophore and a quencher, respectively. When the pair of LCR probes are ligated in LCR, the fluorophore will be brought close to the quencher, and thus, the fluorescence will be specifically quenched by fluorescence resonance energy transfer (FRET). The decrease of fluorescence intensity resulted from FRET can be real-time monitored in the LCR process. With the proposed real-time fluorescence LCR assay, 10 aM DNA targets or 100 pg genomic DNA can be accurately determined and as low as 0.1% mutant DNA can be detected in the presence of a large excess of wild-type DNA, indicating the high sensitivity and specificity. The real-time measuring does not require the detection step after LCR and gives a wide dynamic range for detection of DNA targets (from 10 aM to 1 pM). As LCR has been widely used for detection of SNP, DNA methylation, mRNA and microRNA, the real-time fluorescence LCR assay shows great potential for various genetic analysis. Copyright © 2015 Elsevier B.V. All rights reserved.

Mixed Beam Murine Harderian Gland Tumorigenesis: Predicted Dose-Effect Relationships if neither Synergism nor Antagonism Occurs

DOE Office of Scientific and Technical Information (OSTI.GOV)

Siranart, Nopphon; Blakely, Eleanor A.; Cheng, Alden

Complex mixed radiation fields exist in interplanetary space, and not much is known about their latent effects on space travelers. In silico synergy analysis default predictions are useful when planning relevant mixed-ion-beam experiments and interpreting their results. These predictions are based on individual dose-effect relationships (IDER) for each component of the mixed-ion beam, assuming no synergy or antagonism. For example, a default hypothesis of simple effect additivity has often been used throughout the study of biology. However, for more than a century pharmacologists interested in mixtures of therapeutic drugs have analyzed conceptual, mathematical and practical questions similar to those thatmore » arise when analyzing mixed radiation fields, and have shown that simple effect additivity often gives unreasonable predictions when the IDER are curvilinear. Various alternatives to simple effect additivity proposed in radiobiology, pharmacometrics, toxicology and other fields are also known to have important limitations. In this work, we analyze upcoming murine Harderian gland (HG) tumor prevalence mixed-beam experiments, using customized open-source software and published IDER from past single-ion experiments. The upcoming experiments will use acute irradiation and the mixed beam will include components of high atomic number and energy (HZE). We introduce a new alternative to simple effect additivity, "incremental effect additivity", which is more suitable for the HG analysis and perhaps for other end points. We use incremental effect additivity to calculate default predictions for mixture dose-effect relationships, including 95% confidence intervals. We have drawn three main conclusions from this work. 1. It is important to supplement mixed-beam experiments with single-ion experiments, with matching end point(s), shielding and dose timing. 2. For HG tumorigenesis due to a mixed beam, simple effect additivity and incremental effect additivity sometimes give default predictions that are numerically close. However, if nontargeted effects are important and the mixed beam includes a number of different HZE components, simple effect additivity becomes unusable and another method is needed such as incremental effect additivity. 3. Eventually, synergy analysis default predictions of the effects of mixed radiation fields will be replaced by more mechanistic, biophysically-based predictions. However, optimizing synergy analyses is an important first step. If mixed-beam experiments indicate little synergy or antagonism, plans by NASA for further experiments and possible missions beyond low earth orbit will be substantially simplified.« less

A New Joint-Blade SENSE Reconstruction for Accelerated PROPELLER MRI

PubMed Central

Lyu, Mengye; Liu, Yilong; Xie, Victor B.; Feng, Yanqiu; Guo, Hua; Wu, Ed X.

2017-01-01

PROPELLER technique is widely used in MRI examinations for being motion insensitive, but it prolongs scan time and is restricted mainly to T2 contrast. Parallel imaging can accelerate PROPELLER and enable more flexible contrasts. Here, we propose a multi-step joint-blade (MJB) SENSE reconstruction to reduce the noise amplification in parallel imaging accelerated PROPELLER. MJB SENSE utilizes the fact that PROPELLER blades contain sharable information and blade-combined images can serve as regularization references. It consists of three steps. First, conventional blade-combined images are obtained using the conventional simple single-blade (SSB) SENSE, which reconstructs each blade separately. Second, the blade-combined images are employed as regularization for blade-wise noise reduction. Last, with virtual high-frequency data resampled from the previous step, all blades are jointly reconstructed to form the final images. Simulations were performed to evaluate the proposed MJB SENSE for noise reduction and motion correction. MJB SENSE was also applied to both T2-weighted and T1-weighted in vivo brain data. Compared to SSB SENSE, MJB SENSE greatly reduced the noise amplification at various acceleration factors, leading to increased image SNR in all simulation and in vivo experiments, including T1-weighted imaging with short echo trains. Furthermore, it preserved motion correction capability and was computationally efficient. PMID:28205602

A New Joint-Blade SENSE Reconstruction for Accelerated PROPELLER MRI.

PubMed

Lyu, Mengye; Liu, Yilong; Xie, Victor B; Feng, Yanqiu; Guo, Hua; Wu, Ed X

2017-02-16

PROPELLER technique is widely used in MRI examinations for being motion insensitive, but it prolongs scan time and is restricted mainly to T2 contrast. Parallel imaging can accelerate PROPELLER and enable more flexible contrasts. Here, we propose a multi-step joint-blade (MJB) SENSE reconstruction to reduce the noise amplification in parallel imaging accelerated PROPELLER. MJB SENSE utilizes the fact that PROPELLER blades contain sharable information and blade-combined images can serve as regularization references. It consists of three steps. First, conventional blade-combined images are obtained using the conventional simple single-blade (SSB) SENSE, which reconstructs each blade separately. Second, the blade-combined images are employed as regularization for blade-wise noise reduction. Last, with virtual high-frequency data resampled from the previous step, all blades are jointly reconstructed to form the final images. Simulations were performed to evaluate the proposed MJB SENSE for noise reduction and motion correction. MJB SENSE was also applied to both T2-weighted and T1-weighted in vivo brain data. Compared to SSB SENSE, MJB SENSE greatly reduced the noise amplification at various acceleration factors, leading to increased image SNR in all simulation and in vivo experiments, including T1-weighted imaging with short echo trains. Furthermore, it preserved motion correction capability and was computationally efficient.

Ambulatory estimation of mean step length during unconstrained walking by means of COG accelerometry.

PubMed

González, R C; Alvarez, D; López, A M; Alvarez, J C

2009-12-01

It has been reported that spatio-temporal gait parameters can be estimated using an accelerometer to calculate the vertical displacement of the body's centre of gravity. This method has the potential to produce realistic ambulatory estimations of those parameters during unconstrained walking. In this work, we want to evaluate the crude estimations of mean step length so obtained, for their possible application in the construction of an ambulatory walking distance measurement device. Two methods have been tested with a set of volunteers in 20 m excursions. Experimental results show that estimations of walking distance can be obtained with sufficient accuracy and precision for most practical applications (errors of 3.66 +/- 6.24 and 0.96 +/- 5.55%), the main difficulty being inter-individual variability (biggest deviations of 19.70 and 15.09% for each estimator). Also, the results indicate that an inverted pendulum model for the displacement during the single stance phase, and a constant displacement per step during double stance, constitute a valid model for the travelled distance with no need of further adjustments. It allows us to explain the main part of the erroneous distance estimations in different subjects as caused by fundamental limitations of the simple inverted pendulum approach.

Production of alpha-amylase from Aspergillus oryzae for several industrial applications in a single step.

PubMed

Porfirif, María C; Milatich, Esteban J; Farruggia, Beatriz M; Romanini, Diana

2016-06-01

A one-step method as a strategy of alpha-amylase concentration and purification was developed in this work. This methodology requires the use of a very low concentration of biodegradable polyelectrolyte (Eudragit(®) E-PO) and represents a low cost, fast, easy to scale up and non-polluting technology. Besides, this methodology allows recycling the polymer after precipitation. The formation of reversible soluble/insoluble complexes between alpha-amylase and the polymer Eudragit(®) E-PO was studied, and their precipitation in selected conditions was applied with bioseparation purposes. Turbidimetric assays allowed to determine the pH range where the complexes are insoluble (4.50-7.00); pH 5.50 yielded the highest turbidity of the system. The presence of NaCl (0.05M) in the medium totally dissociates the protein-polymer complexes. When the adequate concentration of polymer was added under these conditions to a liquid culture of Aspergillus oryzae, purification factors of alpha-amylase up to 7.43 and recoveries of 88% were obtained in a simple step without previous clarification. These results demonstrate that this methodology is suitable for the concentration and production of alpha-amylase from this source and could be applied at the beginning of downstream processing. Copyright © 2016 Elsevier B.V. All rights reserved.

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

NASA Astrophysics Data System (ADS)

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.

A novel precursor system and its application to produce tin doped indium oxide.

PubMed

Veith, M; Bubel, C; Zimmer, M

2011-06-14

A new type of precursor has been developed by molecular design and synthesised to produce tin doped indium oxide (ITO). The precursor consists of a newly developed bimetallic indium tin alkoxide, Me(2)In(O(t)Bu)(3)Sn (Me = CH(3), O(t)Bu = OC(CH(3))(3)), which is in equilibrium with an excess of Me(2)In(O(t)Bu). This quasi single-source precursor is applied in a sol-gel process to produce powders and coatings of ITO using a one-step heat treatment process under an inert atmosphere. The main advantage of this system is the simple heat treatment that leads to the disproportionation of the bivalent Sn(II) precursor into Sn(IV) and metallic tin, resulting in an overall reduced state of the metal in the final tin doped indium oxide (ITO) material, hence avoiding the usually necessary reduction step. Solid state (119)Sn-NMR measurements of powder samples confirm the appearance of Sn(II) in an amorphous gel state and of metallic tin after annealing under nitrogen. The corresponding preparation of ITO coatings by spin coating on glass leads to transparent conductive layers with a high transmittance of visible light and a low electrical resistivity without the necessity of a reduction step.

Highly Sensitive Bacteriophage-Based Detection of Brucella abortus in Mixed Culture and Spiked Blood

PubMed Central

Sergueev, Kirill V.; Filippov, Andrey A.; Nikolich, Mikeljon P.

2017-01-01

For decades, bacteriophages (phages) have been used for Brucella species identification in the diagnosis and epidemiology of brucellosis. Traditional Brucella phage typing is a multi-day procedure including the isolation of a pure culture, a step that can take up to three weeks. In this study, we focused on the use of brucellaphages for sensitive detection of the pathogen in clinical and other complex samples, and developed an indirect method of Brucella detection using real-time quantitative PCR monitoring of brucellaphage DNA amplification via replication on live Brucella cells. This assay allowed the detection of single bacteria (down to 1 colony-forming unit per milliliter) within 72 h without DNA extraction and purification steps. The technique was equally efficient with Brucella abortus pure culture and with mixed cultures of B. abortus and α-proteobacterial near neighbors that can be misidentified as Brucella spp., Ochrobactrum anthropi and Afipia felis. The addition of a simple short sample preparation step enabled the indirect phage-based detection of B. abortus in spiked blood, with the same high sensitivity. This indirect phage-based detection assay enables the rapid and sensitive detection of live B. abortus in mixed cultures and in blood samples, and can potentially be applied for detection in other clinical samples and other complex sample types. PMID:28604602

Dynamics of laser-induced damage of spherical nanoparticles by high-intensity ultrashort laser pulses

NASA Astrophysics Data System (ADS)

Komolov, Vladimir L.; Gruzdev, Vitaly E.; Przhibelskii, Sergey G.; Smirnov, Dmitry S.

2012-12-01

Damage of a metal spherical nanoparticle by femtosecond laser pulses is analyzed by splitting the overall process into two steps. The fast step includes electron photoemission from a nanoparticle. It takes place during direct action of a laser pulse and its rate is evaluated as a function of laser and particle parameters by two approaches. Obtained results suggest the formation of significant positive charge of the nanoparticles due to the photoemission. The next step includes ion emission that removes the excessive positive charge and modifies particle structure. It is delayed with respect to the photo-emission and is analyzed by a simple analytical model and modified molecular dynamics. Obtained energy distribution suggests generation of fast ions capable of penetrating into surrounding material and generating defects next to the nanoparticle. The modeling is extended to the case of a nanoparticle on a solid surface to understand the basic mechanism of surface laser damage initiated by nano-contamination. Simulations predict embedding the emitted ions into substrate within a spot with size significantly exceeding the original particle size. We discuss the relation of those effects to the problem of bulk and surface laser-induced damage of optical materials by single and multiple ultrashort laser pulses.

Chaotic He-Ne laser

NASA Astrophysics Data System (ADS)

Kuusela, Tom A.

2017-09-01

A He-Ne laser is an example of a class A laser, which can be described by a single nonlinear differential equation of the complex electric field. This laser system has only one degree of freedom and is thus inherently stable. A He-Ne laser can be driven to the chaotic condition when a large fraction of the output beam is injected back to the laser. In practice, this can be done simply by adding an external mirror. In this situation, the laser system has infinite degrees of freedom and therefore it can have a chaotic attractor. We show the fundamental laser equations and perform elementary stability analysis. In experiments, the laser intensity variations are measured by a simple photodiode circuit. The laser output intensity time series is studied using nonlinear analysis tools which can be found freely on the internet. The results show that the laser system with feedback has an attractor of a reasonably high dimension and that the maximal Lyapunov exponent is positive, which is clear evidence of chaotic behaviour. The experimental setup and analysis steps are so simple that the studies can even be implemented in the undergraduate physics laboratory.

An efficient higher order family of root finders

NASA Astrophysics Data System (ADS)

Petkovic, Ljiljana D.; Rancic, Lidija; Petkovic, Miodrag S.

2008-06-01

A one parameter family of iterative methods for the simultaneous approximation of simple complex zeros of a polynomial, based on a cubically convergent Hansen-Patrick's family, is studied. We show that the convergence of the basic family of the fourth order can be increased to five and six using Newton's and Halley's corrections, respectively. Since these corrections use the already calculated values, the computational efficiency of the accelerated methods is significantly increased. Further acceleration is achieved by applying the Gauss-Seidel approach (single-step mode). One of the most important problems in solving nonlinear equations, the construction of initial conditions which provide both the guaranteed and fast convergence, is considered for the proposed accelerated family. These conditions are computationally verifiable; they depend only on the polynomial coefficients, its degree and initial approximations, which is of practical importance. Some modifications of the considered family, providing the computation of multiple zeros of polynomials and simple zeros of a wide class of analytic functions, are also studied. Numerical examples demonstrate the convergence properties of the presented family of root-finding methods.

Simple and rapid silver nanoparticles based antioxidant capacity assays: Reactivity study for phenolic compounds.

PubMed

Della Pelle, Flavio; Scroccarello, Annalisa; Sergi, Manuel; Mascini, Marcello; Del Carlo, Michele; Compagnone, Dario

2018-08-01

A single-step, rapid (10 min), sensitive silver nanoparticles (AgNPs) based spectrophotometric method for antioxidant capacity (AOC) assay has been developed. The assay is based on the ability of natural polyphenols to reduce Ag(I) and stabilize the produced AgNPs(0) at room temperature. Localized surface plasmon resonance (LSPR) of AgNPs at ≈420 nm is then measured. Using different conditions of pH (8.4) and temperature (45 °C) a further assay based on the production of AgNPs with selectivity for flavonols was also developed. The reactivity of the two AgNPs based assays vs. 15 polyphenols belonging to different chemical classes and 9 different samples has been studied and compared with ABTS, Folin and AuNPs based methods for AOC. The proposed assays had good reproducibility (RSD ≤ 13) and are simple, sensitive and cost effective. Moreover, used in conjunction with the classical AOC assays, can improve the information on the polyphenolic pool of food samples. Copyright © 2018 Elsevier Ltd. All rights reserved.

Tuning the Assembly of Spherical Nanoparticles in Semicrystalline Polymers

NASA Astrophysics Data System (ADS)

Zhao, Dan; Jestin, Jacques; Zhao, Longxi; Kumar, Sanat K.; Mohammadkhani, Mohammad; Benicewicz, Brian C.

We propose a simple, novel strategy to controlling nanoparticle (NPs) dispersion states in a semi-crystalline polymer matrix exploiting the kinetics of polymer crystallization. The system consists of poly(methyl methacrylate) grafted spherical silica NPs and poly(ethylene oxide) matrices, which are thermodynamically miscible in the melt. We first show that no remarkable change was observed in the spatial dispersion of NPs upon fast crystallization. However, for slow crystallization, both TEM and X-ray/neutron scattering reveal that the system starts to be organized in a ``layer-by-layer'' architecture, where the NPs are aligned in the amorphous phases intercalated by the crystalline lamellar phases. More importantly, we have found that the resulting ``sheet-like'' NP morphology gives rise to a 2-fold increase in the storage modulus but without compromising the fracture toughness of the neat polymer. These results open pathways for creating in-situ biomimetic hierarchical structures with improved mechanical properties through a simple, single-step crystallization processing, which could lead to new applications for this largest class of commercially relevant polymeric materials.

Mechanistic evaluation of the pros and cons of digital RT-LAMP for HIV-1 viral load quantification on a microfluidic device and improved efficiency via a two-step digital protocol.

PubMed

Sun, Bing; Shen, Feng; McCalla, Stephanie E; Kreutz, Jason E; Karymov, Mikhail A; Ismagilov, Rustem F

2013-02-05

Here we used a SlipChip microfluidic device to evaluate the performance of digital reverse transcription-loop-mediated isothermal amplification (dRT-LAMP) for quantification of HIV viral RNA. Tests are needed for monitoring HIV viral load to control the emergence of drug resistance and to diagnose acute HIV infections. In resource-limited settings, in vitro measurement of HIV viral load in a simple format is especially needed, and single-molecule counting using a digital format could provide a potential solution. We showed here that when one-step dRT-LAMP is used for quantification of HIV RNA, the digital count is lower than expected and is limited by the yield of desired cDNA. We were able to overcome the limitations by developing a microfluidic protocol to manipulate many single molecules in parallel through a two-step digital process. In the first step we compartmentalize the individual RNA molecules (based on Poisson statistics) and perform reverse transcription on each RNA molecule independently to produce DNA. In the second step, we perform the LAMP amplification on all individual DNA molecules in parallel. Using this new protocol, we increased the absolute efficiency (the ratio between the concentration calculated from the actual count and the expected concentration) of dRT-LAMP 10-fold, from ∼2% to ∼23%, by (i) using a more efficient reverse transcriptase, (ii) introducing RNase H to break up the DNA:RNA hybrid, and (iii) adding only the BIP primer during the RT step. We also used this two-step method to quantify HIV RNA purified from four patient samples and found that in some cases, the quantification results were highly sensitive to the sequence of the patient's HIV RNA. We learned the following three lessons from this work: (i) digital amplification technologies, including dLAMP and dPCR, may give adequate dilution curves and yet have low efficiency, thereby providing quantification values that underestimate the true concentration. Careful validation is essential before a method is considered to provide absolute quantification; (ii) the sensitivity of dLAMP to the sequence of the target nucleic acid necessitates additional validation with patient samples carrying the full spectrum of mutations; (iii) for multistep digital amplification chemistries, such as a combination of reverse transcription with amplification, microfluidic devices may be used to decouple these steps from one another and to perform them under different, individually optimized conditions for improved efficiency.

Combining p-values in replicated single-case experiments with multivariate outcome.

PubMed

Solmi, Francesca; Onghena, Patrick

2014-01-01

Interest in combining probabilities has a long history in the global statistical community. The first steps in this direction were taken by Ronald Fisher, who introduced the idea of combining p-values of independent tests to provide a global decision rule when multiple aspects of a given problem were of interest. An interesting approach to this idea of combining p-values is the one based on permutation theory. The methods belonging to this particular approach exploit the permutation distributions of the tests to be combined, and use a simple function to combine probabilities. Combining p-values finds a very interesting application in the analysis of replicated single-case experiments. In this field the focus, while comparing different treatments effects, is more articulated than when just looking at the means of the different populations. Moreover, it is often of interest to combine the results obtained on the single patients in order to get more global information about the phenomenon under study. This paper gives an overview of how the concept of combining p-values was conceived, and how it can be easily handled via permutation techniques. Finally, the method of combining p-values is applied to a simulated replicated single-case experiment, and a numerical illustration is presented.

Step-to-step spatiotemporal variables and ground reaction forces of intra-individual fastest sprinting in a single session.

PubMed

Nagahara, Ryu; Mizutani, Mirai; Matsuo, Akifumi; Kanehisa, Hiroaki; Fukunaga, Tetsuo

2018-06-01

We aimed to investigate the step-to-step spatiotemporal variables and ground reaction forces during the acceleration phase for characterising intra-individual fastest sprinting within a single session. Step-to-step spatiotemporal variables and ground reaction forces produced by 15 male athletes were measured over a 50-m distance during repeated (three to five) 60-m sprints using a long force platform system. Differences in measured variables between the fastest and slowest trials were examined at each step until the 22nd step using a magnitude-based inferences approach. There were possibly-most likely higher running speed and step frequency (2nd to 22nd steps) and shorter support time (all steps) in the fastest trial than in the slowest trial. Moreover, for the fastest trial there were likely-very likely greater mean propulsive force during the initial four steps and possibly-very likely larger mean net anterior-posterior force until the 17th step. The current results demonstrate that better sprinting performance within a single session is probably achieved by 1) a high step frequency (except the initial step) with short support time at all steps, 2) exerting a greater mean propulsive force during initial acceleration, and 3) producing a greater mean net anterior-posterior force during initial and middle acceleration.

A new simple six-step model to promote recruitment to RCTs was developed and successfully implemented.

PubMed

Realpe, Alba; Adams, Ann; Wall, Peter; Griffin, Damian; Donovan, Jenny L

2016-08-01

How a randomized controlled trial (RCT) is explained to patients is a key determinant of recruitment to that trial. This study developed and implemented a simple six-step model to fully inform patients and to support them in deciding whether to take part or not. Ninety-two consultations with 60 new patients were recorded and analyzed during a pilot RCT comparing surgical and nonsurgical interventions for hip impingement. Recordings were analyzed using techniques of thematic analysis and focused conversation analysis. Early findings supported the development of a simple six-step model to provide a framework for good recruitment practice. Model steps are as follows: (1) explain the condition, (2) reassure patients about receiving treatment, (3) establish uncertainty, (4) explain the study purpose, (5) give a balanced view of treatments, and (6) Explain study procedures. There are also two elements throughout the consultation: (1) responding to patients' concerns and (2) showing confidence. The pilot study was successful, with 70% (n = 60) of patients approached across nine centers agreeing to take part in the RCT, so that the full-scale trial was funded. The six-step model provides a promising framework for successful recruitment to RCTs. Further testing of the model is now required. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Controlling dental enamel-cavity ablation depth with optimized stepping parameters along the focal plane normal using a three axis, numerically controlled picosecond laser.

PubMed

Yuan, Fusong; Lv, Peijun; Wang, Dangxiao; Wang, Lei; Sun, Yuchun; Wang, Yong

2015-02-01

The purpose of this study was to establish a depth-control method in enamel-cavity ablation by optimizing the timing of the focal-plane-normal stepping and the single-step size of a three axis, numerically controlled picosecond laser. Although it has been proposed that picosecond lasers may be used to ablate dental hard tissue, the viability of such a depth-control method in enamel-cavity ablation remains uncertain. Forty-two enamel slices with approximately level surfaces were prepared and subjected to two-dimensional ablation by a picosecond laser. The additive-pulse layer, n, was set to 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70. A three-dimensional microscope was then used to measure the ablation depth, d, to obtain a quantitative function relating n and d. Six enamel slices were then subjected to three dimensional ablation to produce 10 cavities, respectively, with additive-pulse layer and single-step size set to corresponding values. The difference between the theoretical and measured values was calculated for both the cavity depth and the ablation depth of a single step. These were used to determine minimum-difference values for both the additive-pulse layer (n) and single-step size (d). When the additive-pulse layer and the single-step size were set 5 and 45, respectively, the depth error had a minimum of 2.25 μm, and 450 μm deep enamel cavities were produced. When performing three-dimensional ablating of enamel with a picosecond laser, adjusting the timing of the focal-plane-normal stepping and the single-step size allows for the control of ablation-depth error to the order of micrometers.

Hierarchical nanostructured WO3-SnO2 for selective sensing of volatile organic compounds

NASA Astrophysics Data System (ADS)

Nayak, Arpan Kumar; Ghosh, Ruma; Santra, Sumita; Guha, Prasanta Kumar; Pradhan, Debabrata

2015-07-01

It remains a challenge to find a suitable gas sensing material that shows a high response and shows selectivity towards various gases simultaneously. Here, we report a mixed metal oxide WO3-SnO2 nanostructured material synthesized in situ by a simple, single-step, one-pot hydrothermal method at 200 °C in 12 h, and demonstrate its superior sensing behavior towards volatile organic compounds (VOCs) such as ammonia, ethanol and acetone. SnO2 nanoparticles with controlled size and density were uniformly grown on WO3 nanoplates by varying the tin precursor. The density of the SnO2 nanoparticles on the WO3 nanoplates plays a crucial role in the VOC selectivity. The responses of the present mixed metal oxides are found to be much higher than the previously reported results based on single/mixed oxides and noble metal-doped oxides. In addition, the VOC selectivity is found to be highly temperature-dependent, with optimum performance obtained at 200 °C, 300 °C and 350 °C for ammonia, ethanol and acetone, respectively. The present results on the cost-effective noble metal-free WO3-SnO2 sensor could find potential application in human breath analysis by non-invasive detection.It remains a challenge to find a suitable gas sensing material that shows a high response and shows selectivity towards various gases simultaneously. Here, we report a mixed metal oxide WO3-SnO2 nanostructured material synthesized in situ by a simple, single-step, one-pot hydrothermal method at 200 °C in 12 h, and demonstrate its superior sensing behavior towards volatile organic compounds (VOCs) such as ammonia, ethanol and acetone. SnO2 nanoparticles with controlled size and density were uniformly grown on WO3 nanoplates by varying the tin precursor. The density of the SnO2 nanoparticles on the WO3 nanoplates plays a crucial role in the VOC selectivity. The responses of the present mixed metal oxides are found to be much higher than the previously reported results based on single/mixed oxides and noble metal-doped oxides. In addition, the VOC selectivity is found to be highly temperature-dependent, with optimum performance obtained at 200 °C, 300 °C and 350 °C for ammonia, ethanol and acetone, respectively. The present results on the cost-effective noble metal-free WO3-SnO2 sensor could find potential application in human breath analysis by non-invasive detection. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr02571k

The Effects of Multiple-Step and Single-Step Directions on Fourth and Fifth Grade Students' Grammar Assessment Performance

ERIC Educational Resources Information Center

Mazerik, Matthew B.

2006-01-01

The mean scores of English Language Learners (ELL) and English Only (EO) students in 4th and 5th grade (N = 110), across the teacher-administered Grammar Skills Test, were examined for differences in participants' scores on assessments containing single-step directions and assessments containing multiple-step directions. The results indicated no…

Evaluation of accuracy in implant site preparation performed in single- or multi-step drilling procedures.

PubMed

Marheineke, Nadine; Scherer, Uta; Rücker, Martin; von See, Constantin; Rahlf, Björn; Gellrich, Nils-Claudius; Stoetzer, Marcus

2018-06-01

Dental implant failure and insufficient osseointegration are proven results of mechanical and thermal damage during the surgery process. We herein performed a comparative study of a less invasive single-step drilling preparation protocol and a conventional multiple drilling sequence. Accuracy of drilling holes was precisely analyzed and the influence of different levels of expertise of the handlers and additional use of drill template guidance was evaluated. Six experimental groups, deployed in an osseous study model, were representing template-guided and freehanded drilling actions in a stepwise drilling procedure in comparison to a single-drill protocol. Each experimental condition was studied by the drilling actions of respectively three persons without surgical knowledge as well as three highly experienced oral surgeons. Drilling actions were performed and diameters were recorded with a precision measuring instrument. Less experienced operators were able to significantly increase the drilling accuracy using a guiding template, especially when multi-step preparations are performed. Improved accuracy without template guidance was observed when experienced operators were executing single-step versus multi-step technique. Single-step drilling protocols have shown to produce more accurate results than multi-step procedures. The outcome of any protocol can be further improved by use of guiding templates. Operator experience can be a contributing factor. Single-step preparations are less invasive and are promoting osseointegration. Even highly experienced surgeons are achieving higher levels of accuracy by combining this technique with template guidance. Hereby template guidance enables a reduction of hands-on time and side effects during surgery and lead to a more predictable clinical diameter.

Texas two-step: a framework for optimal multi-input single-output deconvolution.

PubMed

Neelamani, Ramesh; Deffenbaugh, Max; Baraniuk, Richard G

2007-11-01

Multi-input single-output deconvolution (MISO-D) aims to extract a deblurred estimate of a target signal from several blurred and noisy observations. This paper develops a new two step framework--Texas Two-Step--to solve MISO-D problems with known blurs. Texas Two-Step first reduces the MISO-D problem to a related single-input single-output deconvolution (SISO-D) problem by invoking the concept of sufficient statistics (SSs) and then solves the simpler SISO-D problem using an appropriate technique. The two-step framework enables new MISO-D techniques (both optimal and suboptimal) based on the rich suite of existing SISO-D techniques. In fact, the properties of SSs imply that a MISO-D algorithm is mean-squared-error optimal if and only if it can be rearranged to conform to the Texas Two-Step framework. Using this insight, we construct new wavelet- and curvelet-based MISO-D algorithms with asymptotically optimal performance. Simulated and real data experiments verify that the framework is indeed effective.

An efficient microwave-assisted synthesis method for the production of water soluble amine-terminated Si nanoparticles.

PubMed

Atkins, Tonya M; Louie, Angelique Y; Kauzlarich, Susan M

2012-07-27

Silicon nanoparticles can be considered a green material, especially when prepared via a microwave-assisted method without the use of highly reactive reducing agents or hydrofluoric acid. A simple solution synthesis of hydrogen-terminated Si- and Mn-doped Si nanoparticles via microwave-assisted synthesis is demonstrated. The reaction of the Zintl salt, Na(4)Si(4), or Mn-doped Na(4)Si(4), Na(4)Si(4(Mn)), with ammonium bromide, NH(4)Br, produces small dispersible nanoparticles along with larger particles that precipitate. Allylamine and 1-amino-10-undecene were reacted with the hydrogen-terminated Si nanoparticles to provide water solubility and stability. A one-pot, single-reaction process and a one-pot, two-step reaction process were investigated. Details of the microwave-assisted process are provided, with the optimal synthesis being the one-pot, two-step reaction procedure and a total time of about 15 min. The nanoparticles were characterized by transmission electron microscopy (TEM), x-ray diffraction, and fluorescence spectroscopies. The microwave-assisted method reliably produces a narrow size distribution of Si nanoparticles in solution.

Core-shell TiO2@ZnO nanorods for efficient ultraviolet photodetection

NASA Astrophysics Data System (ADS)

Panigrahi, Shrabani; Basak, Durga

2011-05-01

Core-shell TiO2@ZnO nanorods (NRs) have been fabricated by a simple two step method: growth of ZnO NRs' array by an aqueous chemical technique and then coating of the NRs with a solution of titanium isopropoxide [Ti(OC3H7)4] followed by a heating step to form the shell. The core-shell nanocomposites are composed of single-crystalline ZnO NRs, coated with a thin TiO2 shell layer obtained by varying the number of coatings (one, three and five times). The ultraviolet (UV) emission intensity of the nanocomposite is largely quenched due to an efficient electron-hole separation reducing the band-to-band recombinations. The UV photoconductivity of the core-shell structure with three times TiO2 coating has been largely enhanced due to photoelectron transfer between the core and the shell. The UV photosensitivity of the nanocomposite becomes four times larger while the photocurrent decay during steady UV illumination has been decreased almost by 7 times compared to the as-grown ZnO NRs indicating high efficiency of these core-shell structures as UV sensors.

Numerical Simulations of Reacting Flows Using Asynchrony-Tolerant Schemes for Exascale Computing

NASA Astrophysics Data System (ADS)

Cleary, Emmet; Konduri, Aditya; Chen, Jacqueline

2017-11-01

Communication and data synchronization between processing elements (PEs) are likely to pose a major challenge in scalability of solvers at the exascale. Recently developed asynchrony-tolerant (AT) finite difference schemes address this issue by relaxing communication and synchronization between PEs at a mathematical level while preserving accuracy, resulting in improved scalability. The performance of these schemes has been validated for simple linear and nonlinear homogeneous PDEs. However, many problems of practical interest are governed by highly nonlinear PDEs with source terms, whose solution may be sensitive to perturbations caused by communication asynchrony. The current work applies the AT schemes to combustion problems with chemical source terms, yielding a stiff system of PDEs with nonlinear source terms highly sensitive to temperature. Examples shown will use single-step and multi-step CH4 mechanisms for 1D premixed and nonpremixed flames. Error analysis will be discussed both in physical and spectral space. Results show that additional errors introduced by the AT schemes are negligible and the schemes preserve their accuracy. We acknowledge funding from the DOE Computational Science Graduate Fellowship administered by the Krell Institute.

Applying the multivariate time-rescaling theorem to neural population models

PubMed Central

Gerhard, Felipe; Haslinger, Robert; Pipa, Gordon

2011-01-01

Statistical models of neural activity are integral to modern neuroscience. Recently, interest has grown in modeling the spiking activity of populations of simultaneously recorded neurons to study the effects of correlations and functional connectivity on neural information processing. However any statistical model must be validated by an appropriate goodness-of-fit test. Kolmogorov-Smirnov tests based upon the time-rescaling theorem have proven to be useful for evaluating point-process-based statistical models of single-neuron spike trains. Here we discuss the extension of the time-rescaling theorem to the multivariate (neural population) case. We show that even in the presence of strong correlations between spike trains, models which neglect couplings between neurons can be erroneously passed by the univariate time-rescaling test. We present the multivariate version of the time-rescaling theorem, and provide a practical step-by-step procedure for applying it towards testing the sufficiency of neural population models. Using several simple analytically tractable models and also more complex simulated and real data sets, we demonstrate that important features of the population activity can only be detected using the multivariate extension of the test. PMID:21395436

40 CFR 35.935-3 - Property.

Code of Federal Regulations, 2010 CFR

2010-07-01

... construction), and must thereafter retain, a fee simple or such estate or interest in the site of a step 3... municipalities have, or will have before the initiation of step 3 construction, such interests or rights in land...

Low-energy transmission electron diffraction and imaging of large-area graphene

PubMed Central

Zhao, Wei; Xia, Bingyu; Lin, Li; Xiao, Xiaoyang; Liu, Peng; Lin, Xiaoyang; Peng, Hailin; Zhu, Yuanmin; Yu, Rong; Lei, Peng; Wang, Jiangtao; Zhang, Lina; Xu, Yong; Zhao, Mingwen; Peng, Lianmao; Li, Qunqing; Duan, Wenhui; Liu, Zhongfan; Fan, Shoushan; Jiang, Kaili

2017-01-01

Two-dimensional (2D) materials have attracted interest because of their excellent properties and potential applications. A key step in realizing industrial applications is to synthesize wafer-scale single-crystal samples. Until now, single-crystal samples, such as graphene domains up to the centimeter scale, have been synthesized. However, a new challenge is to efficiently characterize large-area samples. Currently, the crystalline characterization of these samples still relies on selected-area electron diffraction (SAED) or low-energy electron diffraction (LEED), which is more suitable for characterizing very small local regions. This paper presents a highly efficient characterization technique that adopts a low-energy electrostatically focused electron gun and a super-aligned carbon nanotube (SACNT) film sample support. It allows rapid crystalline characterization of large-area graphene through a single photograph of a transmission-diffracted image at a large beam size. Additionally, the low-energy electron beam enables the observation of a unique diffraction pattern of adsorbates on the suspended graphene at room temperature. This work presents a simple and convenient method for characterizing the macroscopic structures of 2D materials, and the instrument we constructed allows the study of the weak interaction with 2D materials. PMID:28879233

Low-energy transmission electron diffraction and imaging of large-area graphene.

PubMed

Zhao, Wei; Xia, Bingyu; Lin, Li; Xiao, Xiaoyang; Liu, Peng; Lin, Xiaoyang; Peng, Hailin; Zhu, Yuanmin; Yu, Rong; Lei, Peng; Wang, Jiangtao; Zhang, Lina; Xu, Yong; Zhao, Mingwen; Peng, Lianmao; Li, Qunqing; Duan, Wenhui; Liu, Zhongfan; Fan, Shoushan; Jiang, Kaili

2017-09-01

Two-dimensional (2D) materials have attracted interest because of their excellent properties and potential applications. A key step in realizing industrial applications is to synthesize wafer-scale single-crystal samples. Until now, single-crystal samples, such as graphene domains up to the centimeter scale, have been synthesized. However, a new challenge is to efficiently characterize large-area samples. Currently, the crystalline characterization of these samples still relies on selected-area electron diffraction (SAED) or low-energy electron diffraction (LEED), which is more suitable for characterizing very small local regions. This paper presents a highly efficient characterization technique that adopts a low-energy electrostatically focused electron gun and a super-aligned carbon nanotube (SACNT) film sample support. It allows rapid crystalline characterization of large-area graphene through a single photograph of a transmission-diffracted image at a large beam size. Additionally, the low-energy electron beam enables the observation of a unique diffraction pattern of adsorbates on the suspended graphene at room temperature. This work presents a simple and convenient method for characterizing the macroscopic structures of 2D materials, and the instrument we constructed allows the study of the weak interaction with 2D materials.

A simple solution to systematic errors in density determination by X-ray reflectivity: The XRR-density evaluation (XRR-DE) method

NASA Astrophysics Data System (ADS)

Bergese, P.; Bontempi, E.; Depero, L. E.

2006-10-01

X-ray reflectivity (XRR) is a non-destructive, accurate and fast technique for evaluating film density. Indeed, sample-goniometer alignment is a critical experimental factor and the overriding error source in XRR density determination. With commercial single-wavelength X-ray reflectometers, alignment is difficult to control and strongly depends on the operator. In the present work, the contribution of misalignment on density evaluation error is discussed, and a novel procedure (named XRR-density evaluation or XRR-DE method) to minimize the problem will be presented. The method allows to overcome the alignment step through the extrapolation of the correct density value from appropriate non-specular XRR data sets. This procedure is operator independent and suitable for commercial single-wavelength X-ray reflectometers. To test the XRR-DE method, single crystals of TiO 2 and SrTiO 3 were used. In both cases the determined densities differed from the nominal ones less than 5.5%. Thus, the XRR-DE method can be successfully applied to evaluate the density of thin films for which only optical reflectivity is today used. The advantage is that this method can be considered thickness independent.

A Markovian state-space framework for integrating flexibility into space system design decisions

NASA Astrophysics Data System (ADS)

Lafleur, Jarret M.

The past decades have seen the state of the art in aerospace system design progress from a scope of simple optimization to one including robustness, with the objective of permitting a single system to perform well even in off-nominal future environments. Integrating flexibility, or the capability to easily modify a system after it has been fielded in response to changing environments, into system design represents a further step forward. One challenge in accomplishing this rests in that the decision-maker must consider not only the present system design decision, but also sequential future design and operation decisions. Despite extensive interest in the topic, the state of the art in designing flexibility into aerospace systems, and particularly space systems, tends to be limited to analyses that are qualitative, deterministic, single-objective, and/or limited to consider a single future time period. To address these gaps, this thesis develops a stochastic, multi-objective, and multi-period framework for integrating flexibility into space system design decisions. Central to the framework are five steps. First, system configuration options are identified and costs of switching from one configuration to another are compiled into a cost transition matrix. Second, probabilities that demand on the system will transition from one mission to another are compiled into a mission demand Markov chain. Third, one performance matrix for each design objective is populated to describe how well the identified system configurations perform in each of the identified mission demand environments. The fourth step employs multi-period decision analysis techniques, including Markov decision processes from the field of operations research, to find efficient paths and policies a decision-maker may follow. The final step examines the implications of these paths and policies for the primary goal of informing initial system selection. Overall, this thesis unifies state-centric concepts of flexibility from economics and engineering literature with sequential decision-making techniques from operations research. The end objective of this thesis’ framework and its supporting tools is to enable selection of the next-generation space systems today, tailored to decision-maker budget and performance preferences, that will be best able to adapt and perform in a future of changing environments and requirements. Following extensive theoretical development, the framework and its steps are applied to space system planning problems of (1) DARPA-motivated multiple- or distributed-payload satellite selection and (2) NASA human space exploration architecture selection.

Development of a rapid and sensitive one-step reverse transcription-nested polymerase chain reaction in a single tube using the droplet-polymerase chain reaction machine.

PubMed

Yamaguchi, Akemi; Matsuda, Kazuyuki; Sueki, Akane; Taira, Chiaki; Uehara, Masayuki; Saito, Yasunori; Honda, Takayuki

2015-08-25

Reverse transcription (RT)-nested polymerase chain reaction (PCR) is a time-consuming procedure because it has several handling steps and is associated with the risk of cross-contamination during each step. Therefore, a rapid and sensitive one-step RT-nested PCR was developed that could be performed in a single tube using a droplet-PCR machine. The K562 BCR-ABL mRNA-positive cell line as well as bone marrow aspirates from 5 patients with chronic myelogenous leukemia (CML) and 5 controls without CML were used. We evaluated one-step RT-nested PCR using the droplet-PCR machine. One-step RT-nested PCR performed in a single tube using the droplet-PCR machine enabled the detection of BCR-ABL mRNA within 40min, which was 10(3)-fold superior to conventional RT nested PCR using three steps in separate tubes. The sensitivity of the one-step RT-nested PCR was 0.001%, with sample reactivity comparable to that of the conventional assay. One-step RT-nested PCR was developed using the droplet-PCR machine, which enabled all reactions to be performed in a single tube accurately and rapidly and with high sensitivity. This one-step RT-nested PCR may be applicable to a wide spectrum of genetic tests in clinical laboratories. Copyright © 2015 Elsevier B.V. All rights reserved.

Simple Steps to Save Water

EPA Pesticide Factsheets

Saving water around the home is simple and smart. The average household spends as much as $500 per year on its water and sewer bill but could save about $170 per year by retrofitting with waterefficient fixtures and incorporating watersaving practices.

Characteristics of camel-gate structures with active doping channel profiles

NASA Astrophysics Data System (ADS)

Tsai, Jung-Hui; Lour, Wen-Shiung; Laih, Lih-Wen; Liu, Rong-Chau; Liu, Wen-Chau

1996-03-01

In this paper, we demonstrate the influence of channel doping profile on the performances of camel-gate field effect transistors (CAMFETs). For comparison, single and tri-step doping channel structures with identical doping thickness products are employed, while other parameters are kept unchanged. The results of a theoretical analysis show that the single doping channel FET with lightly doping active layer has higher barrier height and drain-source saturation current. However, the transconductance is decreased. For a tri-step doping channel structure, it is found that the output drain-source saturation current and the barrier height are enhanced. Furthermore, the relatively voltage independent performances are improved. Two CAMFETs with single and tri-step doping channel structures have been fabricated and discussed. The devices exhibit nearly voltage independent transconductances of 144 mS mm -1 and 222 mS mm -1 for single and tri-step doping channel CAMFETs, respectively. The operation gate voltage may extend to ± 1.5 V for a tri-step doping channel CAMFET. In addition, the drain current densities of > 750 and 405 mA mm -1 are obtained for the tri-step and single doping CAMFETs. These experimental results are inconsistent with theoretical analysis.

A highly sensitive and accurate gene expression analysis by sequencing ("bead-seq") for a single cell.

PubMed

Matsunaga, Hiroko; Goto, Mari; Arikawa, Koji; Shirai, Masataka; Tsunoda, Hiroyuki; Huang, Huan; Kambara, Hideki

2015-02-15

Analyses of gene expressions in single cells are important for understanding detailed biological phenomena. Here, a highly sensitive and accurate method by sequencing (called "bead-seq") to obtain a whole gene expression profile for a single cell is proposed. A key feature of the method is to use a complementary DNA (cDNA) library on magnetic beads, which enables adding washing steps to remove residual reagents in a sample preparation process. By adding the washing steps, the next steps can be carried out under the optimal conditions without losing cDNAs. Error sources were carefully evaluated to conclude that the first several steps were the key steps. It is demonstrated that bead-seq is superior to the conventional methods for single-cell gene expression analyses in terms of reproducibility, quantitative accuracy, and biases caused during sample preparation and sequencing processes. Copyright © 2014 Elsevier Inc. All rights reserved.

The Story of the Calvin Cycle: Bringing Carbon Fixation to Life

ERIC Educational Resources Information Center

Firooznia, Fardad

2007-01-01

A fun, simple, musical role-playing exercise that allows students to actively step through and visualize the biochemical steps of the Calvin cycle. This musical can easily be completed in about 15 minutes with more than enough time during a 50-minute class period to review the steps and clarify the process further.

Seven Steps You Can Take to Improve Your Objectives. Third Edition.

ERIC Educational Resources Information Center

Alvir, Claire Gelinas

This document gives examples of seven steps a classroom teacher can take to improve both instructional and learning objectives. This improvement is to be measured by increased student learning. The seven steps are as follows: a) write a simple behavioral objective, b) edit, c) revise objective to make it learner centered, d) clarify, e) evaluate…

3 Steps to Great Coaching: A Simple but Powerful Instructional Coaching Cycle Nets Results

ERIC Educational Resources Information Center

Knight, Jim; Elford, Marti; Hock, Michael; Dunekack, Devona; Bradley, Barbara; Deshler, Donald D.; Knight, David

2015-01-01

In this article the authors describe a three-step instructional coaching cycle that can helps coaches become more effective. The article provides the steps and related components to: (1) Identify; (2) Learn; and (3) Improve. While the instructional coaching cycle is only one effective coaching program, coaches also need professional learning that…

Enhancement of orientation gradients during simple shear deformation by application of simple compression

NASA Astrophysics Data System (ADS)

Jahedi, Mohammad; Ardeljan, Milan; Beyerlein, Irene J.; Paydar, Mohammad Hossein; Knezevic, Marko

2015-06-01

We use a multi-scale, polycrystal plasticity micromechanics model to study the development of orientation gradients within crystals deforming by slip. At the largest scale, the model is a full-field crystal plasticity finite element model with explicit 3D grain structures created by DREAM.3D, and at the finest scale, at each integration point, slip is governed by a dislocation density based hardening law. For deformed polycrystals, the model predicts intra-granular misorientation distributions that follow well the scaling law seen experimentally by Hughes et al., Acta Mater. 45(1), 105-112 (1997), independent of strain level and deformation mode. We reveal that the application of a simple compression step prior to simple shearing significantly enhances the development of intra-granular misorientations compared to simple shearing alone for the same amount of total strain. We rationalize that the changes in crystallographic orientation and shape evolution when going from simple compression to simple shearing increase the local heterogeneity in slip, leading to the boost in intra-granular misorientation development. In addition, the analysis finds that simple compression introduces additional crystal orientations that are prone to developing intra-granular misorientations, which also help to increase intra-granular misorientations. Many metal working techniques for refining grain sizes involve a preliminary or concurrent application of compression with severe simple shearing. Our finding reveals that a pre-compression deformation step can, in fact, serve as another processing variable for improving the rate of grain refinement during the simple shearing of polycrystalline metals.

Single-item measures for depression and anxiety: Validation of the Screening Tool for Psychological Distress in an inpatient cardiology setting.

PubMed

Young, Quincy-Robyn; Nguyen, Michelle; Roth, Susan; Broadberry, Ann; Mackay, Martha H

2015-12-01

Depression and anxiety are common among patients with cardiovascular disease (CVD) and confer significant cardiac risk, contributing to CVD morbidity and mortality. Unfortunately, due to the lack of screening tools that address the specific needs of hospitalized patients, few cardiac inpatient programs offer routine screening for these forms of psychological distress, despite recommendations to do so. The purpose of this study was to validate single-item measures for depression and anxiety among cardiac inpatients. Consecutive inpatients were recruited from the cardiology and cardiac surgery step-down units at a university-affiliated, quaternary-care hospital. Subjects completed a questionnaire that included: (a) demographics, (b) single-item-measures for depression and anxiety (from the Screening Tool for Psychological Distress (STOP-D)), and (c) Hospital Anxiety and Depression Scale (HADS). One hundred and five participants were recruited with a wide variety of cardiac diagnoses, having a mean age of 66 years, and 28% were women. Both STOP-D items were highly correlated with their corresponding validated measures and demonstrated robust receiver-operator characteristic curves. Severity scores on both items correlated well with established severity cut-off scores on the corresponding subscales of the HADS. The STOP-D is a self-administered, self-report measure using two independent items that provide severity scores for depression and anxiety. The tool performs very well compared with other previously validated measures. Requiring no additional scoring and being free, STOP-D offers a simple and valid method for identifying hospitalized cardiac patients who are experiencing psychological distress. This crucial first step triggers initiation of appropriate monitoring and intervention, thus reducing the likelihood of the adverse cardiac outcomes associated with psychological distress. © The European Society of Cardiology 2014.

Reward maximization justifies the transition from sensory selection at childhood to sensory integration at adulthood.

PubMed

Daee, Pedram; Mirian, Maryam S; Ahmadabadi, Majid Nili

2014-01-01

In a multisensory task, human adults integrate information from different sensory modalities--behaviorally in an optimal Bayesian fashion--while children mostly rely on a single sensor modality for decision making. The reason behind this change of behavior over age and the process behind learning the required statistics for optimal integration are still unclear and have not been justified by the conventional Bayesian modeling. We propose an interactive multisensory learning framework without making any prior assumptions about the sensory models. In this framework, learning in every modality and in their joint space is done in parallel using a single-step reinforcement learning method. A simple statistical test on confidence intervals on the mean of reward distributions is used to select the most informative source of information among the individual modalities and the joint space. Analyses of the method and the simulation results on a multimodal localization task show that the learning system autonomously starts with sensory selection and gradually switches to sensory integration. This is because, relying more on modalities--i.e. selection--at early learning steps (childhood) is more rewarding than favoring decisions learned in the joint space since, smaller state-space in modalities results in faster learning in every individual modality. In contrast, after gaining sufficient experiences (adulthood), the quality of learning in the joint space matures while learning in modalities suffers from insufficient accuracy due to perceptual aliasing. It results in tighter confidence interval for the joint space and consequently causes a smooth shift from selection to integration. It suggests that sensory selection and integration are emergent behavior and both are outputs of a single reward maximization process; i.e. the transition is not a preprogrammed phenomenon.

Efficient and simpler method to construct normalized cDNA libraries with improved representations of full-length cDNAs

DOEpatents

Soares, Marcelo Bento; Bonaldo, Maria de Fatima

1998-01-01

This invention provides a method to normalize a cDNA library comprising: (a) constructing a directionally cloned library containing cDNA inserts wherein the insert is capable of being amplified by polymerase chain reaction; (b) converting a double-stranded cDNA library into single-stranded DNA circles; (c) generating single-stranded nucleic acid molecules complementary to the single-stranded DNA circles converted in step (b) by polymerase chain reaction with appropriate primers; (d) hybridizing the single-stranded DNA circles converted in step (b) with the complementary single-stranded nucleic acid molecules generated in step (c) to produce partial duplexes to an appropriate Cot; and (e) separating the unhybridized single-stranded DNA circles from the hybridized DNA circles, thereby generating a normalized cDNA library. This invention also provides a method to normalize a cDNA library wherein the generating of single-stranded nucleic acid molecules complementary to the single-stranded DNA circles converted in step (b) is by excising cDNA inserts from the double-stranded cDNA library; purifying the cDNA inserts from cloning vectors; and digesting the cDNA inserts with an exonuclease. This invention further provides a method to construct a subtractive cDNA library following the steps described above. This invention further provides normalized and/or subtractive cDNA libraries generated by the above methods.

Efficient and simpler method to construct normalized cDNA libraries with improved representations of full-length cDNAs

DOEpatents

Soares, M.B.; Fatima Bonaldo, M. de

1998-12-08

This invention provides a method to normalize a cDNA library comprising: (a) constructing a directionally cloned library containing cDNA inserts wherein the insert is capable of being amplified by polymerase chain reaction; (b) converting a double-stranded cDNA library into single-stranded DNA circles; (c) generating single-stranded nucleic acid molecules complementary to the single-stranded DNA circles converted in step (b) by polymerase chain reaction with appropriate primers; (d) hybridizing the single-stranded DNA circles converted in step (b) with the complementary single-stranded nucleic acid molecules generated in step (c) to produce partial duplexes to an appropriate Cot; and (e) separating the unhybridized single-stranded DNA circles from the hybridized DNA circles, thereby generating a normalized cDNA library. This invention also provides a method to normalize a cDNA library wherein the generating of single-stranded nucleic acid molecules complementary to the single-stranded DNA circles converted in step (b) is by excising cDNA inserts from the double-stranded cDNA library; purifying the cDNA inserts from cloning vectors; and digesting the cDNA inserts with an exonuclease. This invention further provides a method to construct a subtractive cDNA library following the steps described above. This invention further provides normalized and/or subtractive cDNA libraries generated by the above methods. 25 figs.

Increased knee valgus alignment and moment during single-leg landing after overhead stroke as a potential risk factor of anterior cruciate ligament injury in badminton.

PubMed

Kimura, Yuka; Ishibashi, Yasuyuki; Tsuda, Eiichi; Yamamoto, Yuji; Hayashi, Yoshimitsu; Sato, Shuichi

2012-03-01

In badminton, knees opposite to the racket-hand side received anterior cruciate ligament (ACL) injuries during single-leg landing after overhead stroke. Most of them occurred in the backhand-side of the rear court. Comparing lower limb biomechanics during single-leg landing after overhead stroke between the forehand-side and backhand-side court may help explain the different injury rates depending on court position. The knee kinematics and kinetics during single-leg landing after overhead stroke following back-stepping were different between the forehand-side and backhand-side court. Controlled laboratory study. Hip, knee and ankle joint kinematic and knee kinetic data were collected for 17 right-handed female college badminton players using a 3-dimensional motion analysis system. Subjects performed single-left-legged landing after an overhead stroke following left and right back-stepping. The kinematic and kinetic data of the left lower extremities during landing were measured and compared between left and right back-steps. Hip flexion and abduction and knee valgus at the initial contact, hip and knee flexion and knee valgus at the maximum knee flexion and the maximum knee valgus moment were significantly larger for the left back-step than the right back-step (p<0.05). Significant differences in joint kinematics and kinetics of the lower extremity during single-leg landing after overhead stroke were observed between different back-step directions. Increased knee valgus angle and moment following back-stepping to the backhand-side might be related to the higher incidence of ACL injury during single-leg landing after overhead stroke.

Single-step reinitialization and extending algorithms for level-set based multi-phase flow simulations

NASA Astrophysics Data System (ADS)

Fu, Lin; Hu, Xiangyu Y.; Adams, Nikolaus A.

2017-12-01

We propose efficient single-step formulations for reinitialization and extending algorithms, which are critical components of level-set based interface-tracking methods. The level-set field is reinitialized with a single-step (non iterative) "forward tracing" algorithm. A minimum set of cells is defined that describes the interface, and reinitialization employs only data from these cells. Fluid states are extrapolated or extended across the interface by a single-step "backward tracing" algorithm. Both algorithms, which are motivated by analogy to ray-tracing, avoid multiple block-boundary data exchanges that are inevitable for iterative reinitialization and extending approaches within a parallel-computing environment. The single-step algorithms are combined with a multi-resolution conservative sharp-interface method and validated by a wide range of benchmark test cases. We demonstrate that the proposed reinitialization method achieves second-order accuracy in conserving the volume of each phase. The interface location is invariant to reapplication of the single-step reinitialization. Generally, we observe smaller absolute errors than for standard iterative reinitialization on the same grid. The computational efficiency is higher than for the standard and typical high-order iterative reinitialization methods. We observe a 2- to 6-times efficiency improvement over the standard method for serial execution. The proposed single-step extending algorithm, which is commonly employed for assigning data to ghost cells with ghost-fluid or conservative interface interaction methods, shows about 10-times efficiency improvement over the standard method while maintaining same accuracy. Despite their simplicity, the proposed algorithms offer an efficient and robust alternative to iterative reinitialization and extending methods for level-set based multi-phase simulations.

Simple and rapid purification of pediocin PA-1 from Pediococcus pentosaceous NCDC 273 suitable for industrial application.

PubMed

Vijay Simha, B; Sood, S K; Kumariya, Rashmi; Garsa, Anita Kumari

2012-10-12

The use of pediocins as food additives or drugs requires a simple and rapid method by which large quantities of homogeneous pediocin are produced at industrial level. Two centrifugation steps required during initial stages of purification i.e. separation of cells from fermentation broth and collection of precipitates after ammonium sulphate precipitation are the major bottlenecks for their large scale purification. In the present work, pediocin production by a new a dairy strain, Pediococcus pentosaceous NCDC 273 (identical to pediocin PA-1 at nucleotide sequence level), was found to be optimum at initial pH of 6.0 and 7.0 of basal MRS supplemented with 20 g/l of glucose or lactose at 20 and 24 h, respectively. Immobilization of cells through entrapment in alginate-xanthan gum gel beads with chitosan coating resulted in negligible cell release during fermentation. Thus, the cell free extract was directly collected through decantation, avoiding the need of centrifugation step at this stage. Subsequent ammonium sulphate precipitation at isoelectric point of pediocin PA-1 (8.85), using magnetic stirrer at high speed (approx. 1200 rpm), resulted in forceful deposition of precipitates on the wall of precipitation beaker allowing their collection using a spatula, avoiding centrifugation step at this stage also. Further purification using cation-exchange chromatography resulted in yield of 134.4% with more than 320 fold purification with the specific activity of 19×10⁵ AU/mg. The collection of single peak of pediocin at 41.9min in RP-HPLC, overlapping with standard pediocin PA-1, resulted in yield of 1.15 μg from 20 μl of sample applied. The overlapping of RP-HPLC peak and SDS-PAGE band corresponding to 4.6 kDa, confirmed the purity and identity of pediocin 273 as pediocin PA-1. Copyright © 2012 Elsevier GmbH. All rights reserved.

Dry contact transfer printing of aligned carbon nanotube patterns and characterization of their optical properties for diameter distribution and alignment.

PubMed

Pint, Cary L; Xu, Ya-Qiong; Moghazy, Sharief; Cherukuri, Tonya; Alvarez, Noe T; Haroz, Erik H; Mahzooni, Salma; Doorn, Stephen K; Kono, Junichiro; Pasquali, Matteo; Hauge, Robert H

2010-02-23

A scalable and facile approach is demonstrated where as-grown patterns of well-aligned structures composed of single-walled carbon nanotubes (SWNT) synthesized via water-assisted chemical vapor deposition (CVD) can be transferred, or printed, to any host surface in a single dry, room-temperature step using the growth substrate as a stamp. We demonstrate compatibility of this process with multiple transfers for large-scale device and specifically tailored pattern fabrication. Utilizing this transfer approach, anisotropic optical properties of the SWNT films are probed via polarized absorption, Raman, and photoluminescence spectroscopies. Using a simple model to describe optical transitions in the large SWNT species present in the aligned samples, polarized absorption data are demonstrated as an effective tool for accurate assignment of the diameter distribution from broad absorption features located in the infrared. This can be performed on either well-aligned samples or unaligned doped samples, allowing simple and rapid feedback of the SWNT diameter distribution that can be challenging and time-consuming to obtain in other optical methods. Furthermore, we discuss challenges in accurately characterizing alignment in structures of long versus short carbon nanotubes through optical techniques, where SWNT length makes a difference in the information obtained in such measurements. This work provides new insight to the efficient transfer and optical properties of an emerging class of long, large diameter SWNT species typically produced in the CVD process.

A Paper-Based Device for Performing Loop-Mediated Isothermal Amplification with Real-Time Simultaneous Detection of Multiple DNA Targets.

PubMed

Seok, Youngung; Joung, Hyou-Arm; Byun, Ju-Young; Jeon, Hyo-Sung; Shin, Su Jeong; Kim, Sanghyo; Shin, Young-Beom; Han, Hyung Soo; Kim, Min-Gon

2017-01-01

Paper-based diagnostic devices have many advantages as a one of the multiple diagnostic test platforms for point-of-care (POC) testing because they have simplicity, portability, and cost-effectiveness. However, despite high sensitivity and specificity of nucleic acid testing (NAT), the development of NAT based on a paper platform has not progressed as much as the others because various specific conditions for nucleic acid amplification reactions such as pH, buffer components, and temperature, inhibitions from technical differences of paper-based device. Here, we propose a paper-based device for performing loop-mediated isothermal amplification (LAMP) with real-time simultaneous detection of multiple DNA targets. We determined the optimal chemical components to enable dry conditions for the LAMP reaction without lyophilization or other techniques. We also devised the simple paper device structure by sequentially stacking functional layers, and employed a newly discovered property of hydroxynaphthol blue fluorescence to analyze real-time LAMP signals in the paper device. This proposed platform allowed analysis of three different meningitis DNA samples in a single device with single-step operation. This LAMP-based multiple diagnostic device has potential for real-time analysis with quantitative detection of 10 2 -10 5 copies of genomic DNA. Furthermore, we propose the transformation of DNA amplification devices to a simple and affordable paper system approach with great potential for realizing a paper-based NAT system for POC testing.

A Paper-Based Device for Performing Loop-Mediated Isothermal Amplification with Real-Time Simultaneous Detection of Multiple DNA Targets

PubMed Central

Seok, Youngung; Joung, Hyou-Arm; Byun, Ju-Young; Jeon, Hyo-Sung; Shin, Su Jeong; Kim, Sanghyo; Shin, Young-Beom; Han, Hyung Soo; Kim, Min-Gon

2017-01-01

Paper-based diagnostic devices have many advantages as a one of the multiple diagnostic test platforms for point-of-care (POC) testing because they have simplicity, portability, and cost-effectiveness. However, despite high sensitivity and specificity of nucleic acid testing (NAT), the development of NAT based on a paper platform has not progressed as much as the others because various specific conditions for nucleic acid amplification reactions such as pH, buffer components, and temperature, inhibitions from technical differences of paper-based device. Here, we propose a paper-based device for performing loop-mediated isothermal amplification (LAMP) with real-time simultaneous detection of multiple DNA targets. We determined the optimal chemical components to enable dry conditions for the LAMP reaction without lyophilization or other techniques. We also devised the simple paper device structure by sequentially stacking functional layers, and employed a newly discovered property of hydroxynaphthol blue fluorescence to analyze real-time LAMP signals in the paper device. This proposed platform allowed analysis of three different meningitis DNA samples in a single device with single-step operation. This LAMP-based multiple diagnostic device has potential for real-time analysis with quantitative detection of 102-105 copies of genomic DNA. Furthermore, we propose the transformation of DNA amplification devices to a simple and affordable paper system approach with great potential for realizing a paper-based NAT system for POC testing. PMID:28740546

Multifunctional microvalves control by optical illumination on nanoheaters and its application in centrifugal microfluidic devices.

PubMed

Park, Jong-Myeon; Cho, Yoon-Kyoung; Lee, Beom-Seok; Lee, Jeong-Gun; Ko, Christopher

2007-05-01

Valving is critical in microfluidic systems. Among many innovative microvalves used in lab-on-a-chip applications, phase change based microvalves using paraffin wax are particularly attractive for disposable biochip applications because they are simple to implement, cost-effective and biocompatible. However, previously reported paraffin-based valves require embedded microheaters and therefore multi-step operation of many microvalves was a difficult problem. Besides, the operation time was relatively long, 2-10 s. In this paper, we report a unique phase change based microvalve for rapid and versatile operation of multiple microvalves using a single laser diode. The valve is made of nanocomposite materials in which 10 nm-sized iron oxide nanoparticles are dispersed in paraffin wax and used as nanoheaters when excited by laser irradiation. Laser light of relatively weak intensity was able to melt the paraffin wax with the embedded iron oxide nanoparticles, whereas even a very intense laser beam does not melt wax alone. The microvalves are leak-free up to 403.0 +/- 7.6 kPa and the response times to operate both normally closed and normally opened microvalves are less than 0.5 s. Furthermore, a sequential operation of multiple microvalves on a centrifugal microfluidic device using a single laser diode was demonstrated. It showed that the optical control of multiple microvalves is fast, robust, simple to operate, and requires minimal chip space and thus is well suited for fully integrated lab-on-a-chip applications.

Quenching of Unincorporated Amplification Signal Reporters in Reverse-Transcription Loop-Mediated Isothermal Amplification Enabling Bright, Single-Step, Closed-Tube, and Multiplexed Detection of RNA Viruses.

PubMed

Ball, Cameron S; Light, Yooli K; Koh, Chung-Yan; Wheeler, Sarah S; Coffey, Lark L; Meagher, Robert J

2016-04-05

Reverse-transcription-loop-mediated isothermal amplification (RT-LAMP) has frequently been proposed as an enabling technology for simplified diagnostic tests for RNA viruses. However, common detection techniques used for LAMP and RT-LAMP have drawbacks, including poor discrimination capability, inability to multiplex targets, high rates of false positives, and (in some cases) the requirement of opening reaction tubes postamplification. Here, we present a simple technique that allows closed-tube, target-specific detection, based on inclusion of a dye-labeled primer that is incorporated into a target-specific amplicon if the target is present. A short, complementary quencher hybridizes to unincorporated primer upon cooling down at the end of the reaction, thereby quenching fluorescence of any unincorporated primer. Our technique, which we term QUASR (for quenching of unincorporated amplification signal reporters, read "quasar"), does not significantly reduce the amplification efficiency or sensitivity of RT-LAMP. Equipped with a simple LED excitation source and a colored plastic gel filter, the naked eye or a camera can easily discriminate between positive and negative QUASR reactions, which produce a difference in signal of approximately 10:1 without background subtraction. We demonstrate that QUASR detection is compatible with complex sample matrices such as human blood, using a novel LAMP primer set for bacteriophage MS2 (a model RNA virus particle). Furthermore, we demonstrate single-tube duplex detection of West Nile virus (WNV) and chikungunya virus (CHIKV) RNA.

Prevalence of gestational diabetes mellitus according to the different criterias

PubMed Central

Akgöl, Evren; Abuşoğlu, Sedat; Gün, Faik Deniz; Ünlü, Ali

2017-01-01

Objective: The two-step approach recommended by the National Diabetes Data Group (NDDG), Carpenter and Coustan (C&C), and O’Sullivan, and the single-step approach recommended by the International Association of Diabetes and Pregnancy Study Group (IADPSG) are used to diagnose gestational diabetes mellitus (GDM). We aimed to determine GDM prevalence and to compare the two-step and single-step approaches used in the southeastern region of Turkey. Materials and Methods: In total, 3048 records of pregnant women screened for GDM between 2008 and 2014 were retrospectively extracted from our laboratory information system. GDM was defined according to the criteria of NDDG, C&C, and O’Sullivan between in 2008 and 2011, and according to those of the IADPSG between 2012 and 2014. Demographic variables were compared using student’s t-test. The linear trends in GDM prevalence with age were calculated using logistic regression. Results: GDM prevalence was found as 4.8%, 8%, and 13.4% using the NDDG, C&C, and O’Sullivan two-step approach, respectively, and 22.3% with the IADPSG single-step approach. GDM prevalence increased with increasing age in both approaches. Conclusion: GDM prevalence was higher using the single-step approach than with the two-step approach. There was a significant increase in GDM prevalence using the IADPSG criteria. PMID:28913130

Into Thin Air.

ERIC Educational Resources Information Center

Kennedy, Mike

2001-01-01

Shows how schools are working to avoid the types of equipment, supplies, and maintenance practices that harm indoor air quality. Simple steps to maintaining a cleaner indoor air environment are highlighted as are steps to reducing the problem air quality and the occurrence of asthma. (GR)

Solid-phase submonomer synthesis of peptoid polymers and their self-assembly into highly-ordered nanosheets.

PubMed

Tran, Helen; Gael, Sarah L; Connolly, Michael D; Zuckermann, Ronald N

2011-11-02

Peptoids are a novel class of biomimetic, non-natural, sequence-specific heteropolymers that resist proteolysis, exhibit potent biological activity, and fold into higher order nanostructures. Structurally similar to peptides, peptoids are poly N-substituted glycines, where the side chains are attached to the nitrogen rather than the alpha-carbon. Their ease of synthesis and structural diversity allows testing of basic design principles to drive de novo design and engineering of new biologically-active and nanostructured materials. Here, a simple manual peptoid synthesis protocol is presented that allows the synthesis of long chain polypeptoids (up to 50mers) in excellent yields. Only basic equipment, simple techniques (e.g. liquid transfer, filtration), and commercially available reagents are required, making peptoids an accessible addition to many researchers' toolkits. The peptoid backbone is grown one monomer at a time via the submonomer method which consists of a two-step monomer addition cycle: acylation and displacement. First, bromoacetic acid activated in situ with N,N'-diisopropylcarbodiimide acylates a resin-bound secondary amine. Second, nucleophilic displacement of the bromide by a primary amine follows to introduce the side chain. The two-step cycle is iterated until the desired chain length is reached. The coupling efficiency of this two-step cycle routinely exceeds 98% and enables the synthesis of peptoids as long as 50 residues. Highly tunable, precise and chemically diverse sequences are achievable with the submonomer method as hundreds of readily available primary amines can be directly incorporated. Peptoids are emerging as a versatile biomimetic material for nanobioscience research because of their synthetic flexibility, robustness, and ordering at the atomic level. The folding of a single-chain, amphiphilic, information-rich polypeptoid into a highly-ordered nanosheet was recently demonstrated. This peptoid is a 36-mer that consists of only three different commercially available monomers: hydrophobic, cationic and anionic. The hydrophobic phenylethyl side chains are buried in the nanosheet core whereas the ionic amine and carboxyl side chains align on the hydrophilic faces. The peptoid nanosheets serve as a potential platform for membrane mimetics, protein mimetics, device fabrication, and sensors. Methods for peptoid synthesis, sheet formation, and microscopy imaging are described and provide a simple method to enable future peptoid nanosheet designs.

In vivo Assembly in Escherichia coli of Transformation Vectors for Plastid Genome Engineering

PubMed Central

Wu, Yuyong; You, Lili; Li, Shengchun; Ma, Meiqi; Wu, Mengting; Ma, Lixin; Bock, Ralph; Chang, Ling; Zhang, Jiang

2017-01-01

Plastid transformation for the expression of recombinant proteins and entire metabolic pathways has become a promising tool for plant biotechnology. However, large-scale application of this technology has been hindered by some technical bottlenecks, including lack of routine transformation protocols for agronomically important crop plants like rice or maize. Currently, there are no standard or commercial plastid transformation vectors available for the scientific community. Construction of a plastid transformation vector usually requires tedious and time-consuming cloning steps. In this study, we describe the adoption of an in vivo Escherichia coli cloning (iVEC) technology to quickly assemble a plastid transformation vector. The method enables simple and seamless build-up of a complete plastid transformation vector from five DNA fragments in a single step. The vector assembled for demonstration purposes contains an enhanced green fluorescent protein (GFP) expression cassette, in which the gfp transgene is driven by the tobacco plastid ribosomal RNA operon promoter fused to the 5′ untranslated region (UTR) from gene10 of bacteriophage T7 and the transcript-stabilizing 3′UTR from the E. coli ribosomal RNA operon rrnB. Successful transformation of the tobacco plastid genome was verified by Southern blot analysis and seed assays. High-level expression of the GFP reporter in the transplastomic plants was visualized by confocal microscopy and Coomassie staining, and GFP accumulation was ~9% of the total soluble protein. The iVEC method represents a simple and efficient approach for construction of plastid transformation vector, and offers great potential for the assembly of increasingly complex vectors for synthetic biology applications in plastids. PMID:28871270

In vivo Assembly in Escherichia coli of Transformation Vectors for Plastid Genome Engineering.

PubMed

Wu, Yuyong; You, Lili; Li, Shengchun; Ma, Meiqi; Wu, Mengting; Ma, Lixin; Bock, Ralph; Chang, Ling; Zhang, Jiang

2017-01-01

Plastid transformation for the expression of recombinant proteins and entire metabolic pathways has become a promising tool for plant biotechnology. However, large-scale application of this technology has been hindered by some technical bottlenecks, including lack of routine transformation protocols for agronomically important crop plants like rice or maize. Currently, there are no standard or commercial plastid transformation vectors available for the scientific community. Construction of a plastid transformation vector usually requires tedious and time-consuming cloning steps. In this study, we describe the adoption of an in vivo Escherichia coli cloning (iVEC) technology to quickly assemble a plastid transformation vector. The method enables simple and seamless build-up of a complete plastid transformation vector from five DNA fragments in a single step. The vector assembled for demonstration purposes contains an enhanced green fluorescent protein (GFP) expression cassette, in which the gfp transgene is driven by the tobacco plastid ribosomal RNA operon promoter fused to the 5' untranslated region (UTR) from gene10 of bacteriophage T7 and the transcript-stabilizing 3'UTR from the E. coli ribosomal RNA operon rrnB . Successful transformation of the tobacco plastid genome was verified by Southern blot analysis and seed assays. High-level expression of the GFP reporter in the transplastomic plants was visualized by confocal microscopy and Coomassie staining, and GFP accumulation was ~9% of the total soluble protein. The iVEC method represents a simple and efficient approach for construction of plastid transformation vector, and offers great potential for the assembly of increasingly complex vectors for synthetic biology applications in plastids.

A simple rapid process for semi-automated brain extraction from magnetic resonance images of the whole mouse head.

PubMed

Delora, Adam; Gonzales, Aaron; Medina, Christopher S; Mitchell, Adam; Mohed, Abdul Faheem; Jacobs, Russell E; Bearer, Elaine L

2016-01-15

Magnetic resonance imaging (MRI) is a well-developed technique in neuroscience. Limitations in applying MRI to rodent models of neuropsychiatric disorders include the large number of animals required to achieve statistical significance, and the paucity of automation tools for the critical early step in processing, brain extraction, which prepares brain images for alignment and voxel-wise statistics. This novel timesaving automation of template-based brain extraction ("skull-stripping") is capable of quickly and reliably extracting the brain from large numbers of whole head images in a single step. The method is simple to install and requires minimal user interaction. This method is equally applicable to different types of MR images. Results were evaluated with Dice and Jacquard similarity indices and compared in 3D surface projections with other stripping approaches. Statistical comparisons demonstrate that individual variation of brain volumes are preserved. A downloadable software package not otherwise available for extraction of brains from whole head images is included here. This software tool increases speed, can be used with an atlas or a template from within the dataset, and produces masks that need little further refinement. Our new automation can be applied to any MR dataset, since the starting point is a template mask generated specifically for that dataset. The method reliably and rapidly extracts brain images from whole head images, rendering them useable for subsequent analytical processing. This software tool will accelerate the exploitation of mouse models for the investigation of human brain disorders by MRI. Copyright © 2015 Elsevier B.V. All rights reserved.

Quantitative multiresidue analysis of antibiotics in milk and milk powder by ultra-performance liquid chromatography coupled to tandem quadrupole mass spectrometry.

PubMed

Tian, He; Wang, Jiaqi; Zhang, Yangdong; Li, Songli; Jiang, Jindou; Tao, Dali; Zheng, Nan

2016-10-15

A simple and fast multiresidue extraction and purification method was developed for the determination of 61 veterinary drugs, belonging to seven classes, in milk and milk powder. The extraction depends on the acetonitrile solvent, followed by a single step to remove lipids with fatty acid chains using a new reversed phase SPE without traditional pre-equilibration and washing steps before eluting SPE. The purifying lipid effect of the present preparation method was evaluated by comparing the response changes of ion peak areas of the milk endogenous metabolites before and after SPE treatment using ultra-fast LC coupled to tandem quadrupole and TOF MS. Subsequently, UPLC coupled to tandem quadrupole MS was performed for the quantitative analysis of milk and milk powder samples spiked with 61 veterinary drugs, including β-lactam, macrolide, amide alcohol, forest amine, sulfanilamide, tetracyclines, and quinolones antibiotics. This method is very simple, fast, sensitive, and selective, and allows the good recoveries of all compounds, with a recovery range of 61.5-118.6%, and coefficients of variation of less than 11.6%. The 61 compounds behave in the dynamic range 0.01-200μgkg(-1), with correlation coefficient >0.99. The limits of quantification for the analytes are in the range 0.01-5.18μgkg(-1). Finally, this method has been successfully applied to the screening of veterinary drugs in 50 commercial bovine milk and milk powder samples, and ceftiofur and ciprofloxacin were detected in some brand samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Identification of regions of normal grey matter and white matter from pathologic glioblastoma and necrosis in frozen sections using Raman imaging.

PubMed

Kast, Rachel; Auner, Gregory; Yurgelevic, Sally; Broadbent, Brandy; Raghunathan, Aditya; Poisson, Laila M; Mikkelsen, Tom; Rosenblum, Mark L; Kalkanis, Steven N

2015-11-01

In neurosurgical applications, a tool capable of distinguishing grey matter, white matter, and areas of tumor and/or necrosis in near-real time could greatly aid in tumor resection decision making. Raman spectroscopy is a non-destructive spectroscopic technique which provides molecular information about the tissue under examination based on the vibrational properties of the constituent molecules. With careful measurement and data processing, a spatial step and repeat acquisition of Raman spectra can be used to create Raman images. Forty frozen brain tissue sections were imaged in their entirety using a 300-µm-square measurement grid, and two or more regions of interest within each tissue were also imaged using a 25 µm-square step size. Molecular correlates for histologic features of interest were identified within the Raman spectra, and novel imaging algorithms were developed to compare molecular features across multiple tissues. In previous work, the relative concentration of individual biomolecules was imaged. Here, the relative concentrations of 1004, 1300:1344, and 1660 cm(-1), which correspond primarily to protein and lipid content, were simultaneously imaged across all tissues. This provided simple interpretation of boundaries between grey matter, white matter, and diseased tissue, and corresponded with findings from adjacent hematoxylin and eosin-stained sections. This novel, yet simple, multi-channel imaging technique allows clinically-relevant resolution with straightforward molecular interpretation of Raman images not possible by imaging any single peak. This method can be applied to either surgical or laboratory tools for rapid, non-destructive imaging of grey and white matter.

Quality testing of an innovative cascade separation system for multiple cell separation

NASA Astrophysics Data System (ADS)

Pierzchalski, Arkadiusz; Moszczynska, Aleksandra; Albrecht, Bernd; Heinrich, Jan-Michael; Tarnok, Attila

2012-03-01

Isolation of different cell types from mixed samples in one separation step by FACS is feasible but expensive and slow. It is cheaper and faster but still challenging by magnetic separation. An innovative bead-based cascade-system (pluriSelect GmbH, Leipzig, Germany) relies on simultaneous physical separation of different cell types. It is based on antibody-mediated binding of cells to beads of different size and isolation with sieves of different mesh-size. We validated pluriSelect system for single parameter (CD3) and simultaneous separation of CD3 and CD15 cells from EDTA blood-samples. Results were compared with those obtained by MACS (Miltenyi-Biotech) magnetic separation (CD3 separation). pluriSelect separation was done in whole blood, MACS on Ficoll gradient isolated leukocytes, according to the manufacturer's protocols. Isolated and residual cells were immunophenotyped (7-color 8-antibody panel (CD3; CD16/56; CD4; CD8; CD14; CD19; CD45; HLADR) on a CyFlowML flow cytometer (Partec GmbH). Cell count (Coulter), purity, yield and viability (7-AAD exclusion) were determined. There were no significant differences between both systems regarding purity (92-98%), yield (50-60%) and viability (92-98%) of isolated cells. PluriSelect separation was slightly faster than MACS (1.15 h versus 1.5h). Moreover, no preenrichment steps were necessary. In conclusion, pluriSelect is a fast, simple and gentle system for efficient simultaneous separation of two cell subpopulation directly from whole blood and can provide a simple alternative to FACS. The isolated cells can be used for further research applications.

Use of aluminum phosphate as the dehydration catalyst in single step dimethyl ether process

DOEpatents

Peng, Xiang-Dong; Parris, Gene E.; Toseland, Bernard A.; Battavio, Paula J.

1998-01-01

The present invention pertains to a process for the coproduction of methanol and dimethyl ether (DME) directly from a synthesis gas in a single step (hereafter, the "single step DME process"). In this process, the synthesis gas comprising hydrogen and carbon oxides is contacted with a dual catalyst system comprising a physical mixture of a methanol synthesis catalyst and a methanol dehydration catalyst. The present invention is an improvement to this process for providing an active and stable catalyst system. The improvement comprises the use of an aluminum phosphate based catalyst as the methanol dehydration catalyst. Due to its moderate acidity, such a catalyst avoids the coke formation and catalyst interaction problems associated with the conventional dual catalyst systems taught for the single step DME process.

Modelface: an Application Programming Interface (API) for Homology Modeling Studies Using Modeller Software

PubMed Central

Sakhteman, Amirhossein; Zare, Bijan

2016-01-01

An interactive application, Modelface, was presented for Modeller software based on windows platform. The application is able to run all steps of homology modeling including pdb to fasta generation, running clustal, model building and loop refinement. Other modules of modeler including energy calculation, energy minimization and the ability to make single point mutations in the PDB structures are also implemented inside Modelface. The API is a simple batch based application with no memory occupation and is free of charge for academic use. The application is also able to repair missing atom types in the PDB structures making it suitable for many molecular modeling studies such as docking and molecular dynamic simulation. Some successful instances of modeling studies using Modelface are also reported. PMID:28243276

Determination of Total Biotin by Liquid Chromatography Coupled with Immunoaffinity Column Cleanup Extraction: Multilaboratory Testing, Final Action 2016.02.

PubMed

Joseph, George; Devi, Ranjani; Marley, Elaine C; Leeman, David

2018-05-01

Single- and multilaboratory testing data have provided systematic scientific evidence that a simple, selective, accurate, and precise method can be used as a potential candidate reference method for dispute resolution in determining total biotin in all forms of infant, adult, and/or pediatric formula. Using LC coupled with immunoaffinity column cleanup extraction, the method fully meets the intended purpose and applicability statement in AOAC Standard Method Performance Requirement 2014.005. The method was applied to a cross-section of infant formula and adult nutritional matrixes, and acceptable precision and accuracy were established. The analytical platform is inexpensive, and the method can be used in almost any laboratory worldwide with basic facilities. The immunoaffinity column cleanup extraction is the key step to successful analysis.

A rapid and simple method for estimating sulfate reduction activity and quantifying inorganic sulfides

USGS Publications Warehouse

Ulrich, G.A.; Krumholz, L.R.; Suflita, J.M.

1997-01-01

A simplified passive extraction procedure for quantifying reduced inorganic sulfur compounds from sediments and water is presented. This method may also be used for the estimation of sulfate reduction rates. Efficient extraction of FeS, FeS(inf2), and S(sup2-) was obtained with this procedure; however, the efficiency for S(sup0) depended on the form that was tested. Passive extraction can be used with samples containing up to 20 mg of reduced sulfur. We demonstrated the utility of this technique in a determination of both sulfate reduction rates and reduced inorganic sulfur pools in marine and freshwater sediments. A side-by-side comparison of the passive extraction method with the established single-step distillation technique yielded comparable results with a fraction of the effort.

The extended Beer-Lambert theory for ray tracing modeling of LED chip-scaled packaging application with multiple luminescence materials

NASA Astrophysics Data System (ADS)

Yuan, Cadmus C. A.

2015-12-01

Optical ray tracing modeling applied Beer-Lambert method in the single luminescence material system to model the white light pattern from blue LED light source. This paper extends such algorithm to a mixed multiple luminescence material system by introducing the equivalent excitation and emission spectrum of individual luminescence materials. The quantum efficiency numbers of individual material and self-absorption of the multiple luminescence material system are considered as well. By this combination, researchers are able to model the luminescence characteristics of LED chip-scaled packaging (CSP), which provides simple process steps and the freedom of the luminescence material geometrical dimension. The method will be first validated by the experimental results. Afterward, a further parametric investigation has been then conducted.

SnAP reagents for the one-step synthesis of medium-ring saturated N-heterocycles from aldehydes

NASA Astrophysics Data System (ADS)

Vo, Cam-Van T.; Luescher, Michael U.; Bode, Jeffrey W.

2014-04-01

Interest in saturated N-heterocycles as scaffolds for the synthesis of bioactive molecules is increasing. Reliable and predictable synthetic methods for the preparation of these compounds, especially medium-sized rings, are limited. We describe the development of SnAP (Sn amino protocol) reagents for the transformation of aldehydes into seven-, eight- and nine-membered saturated N-heterocycles. This process occurs under mild, room-temperature conditions and offers exceptional substrate scope and functional-group tolerance. Air- and moisture-stable SnAP reagents are prepared on a multigram scale from inexpensive starting materials by simple reaction sequences. These new reagents and processes allow widely available aryl, heteroaryl and aliphatic aldehydes to be converted into diverse N-heterocycles, including diazepanes, oxazepanes, diazocanes, oxazocanes and hexahydrobenzoxazonines, by a single synthetic operation.

Optical Properties of CdS Nanobelts and Nanosaws Synthesized by Thermal Evaporation Method

NASA Astrophysics Data System (ADS)

Peng, Zhi-wei; Zou, Bing-suo

2012-04-01

By a simple one-step H2-assisted thermal evaporation method, high quality CdS nanostructures have been successfully fabricated on Au coated Si substrates in large scale. The as-synthesized CdS nanostructures consisted of sword-like nanobelts and toothed nanosaws with a single-crystal hexagonal wurtzite structure. The deposition temperature played an important role in determining the size and morphology of the CdS nanostructures. A combination of vapor-liquid-solid and vapor-solid growth mechanisms were proposed to interpret the formation of CdS nanostructures. Photoluminescence measurement indicated that the nanobelts and nanosaws have a prominent green emission at about 512 nm, which is the band-to-band emission of CdS. The waveguide characteristics of both types of CdS nanostructures were observed and discussed.

Fabrication of chitosan-silver nanoparticle hybrid 3D porous structure as a SERS substrate for biomedical applications

NASA Astrophysics Data System (ADS)

Jung, Gyeong-Bok; Kim, Ji-Hye; Burm, Jin Sik; Park, Hun-Kuk

2013-05-01

We propose a simple, low-cost, large-area, and functional surface enhanced Raman scattering (SERS) substrate for biomedical applications. The SERS substrate with chitosan-silver nanoparticles (chitosan-Ag NPs) hybrid 3D porous structure was fabricated simply by a one-step method. The chitosan was used as a template for the Ag NPs deposition. SERS enhancement by the chitosan-Ag NPs substrate was experimentally verified using rhodamine B as an analyte. Thiolated single stranded DNA was also measured for atopic dermatitis genetic markers (chemokines CCL17) at a low concentration of 5 pM. We successfully designed a novel SERS substrate with silver nanoparticle hybridized 3D porous chitosan that has the potential to become a highly sensitive and selective tool for biomedical applications.

Photochemical Approaches to Complex Chemotypes: Applications in Natural Product Synthesis.

PubMed

Kärkäs, Markus D; Porco, John A; Stephenson, Corey R J

2016-09-14

The use of photochemical transformations is a powerful strategy that allows for the formation of a high degree of molecular complexity from relatively simple building blocks in a single step. A central feature of all light-promoted transformations is the involvement of electronically excited states, generated upon absorption of photons. This produces transient reactive intermediates and significantly alters the reactivity of a chemical compound. The input of energy provided by light thus offers a means to produce strained and unique target compounds that cannot be assembled using thermal protocols. This review aims at highlighting photochemical transformations as a tool for rapidly accessing structurally and stereochemically diverse scaffolds. Synthetic designs based on photochemical transformations have the potential to afford complex polycyclic carbon skeletons with impressive efficiency, which are of high value in total synthesis.

Voluntary stepping behavior under single- and dual-task conditions in chronic stroke survivors: A comparison between the involved and uninvolved legs.

PubMed

Melzer, Itshak; Goldring, Melissa; Melzer, Yehudit; Green, Elad; Tzedek, Irit

2010-12-01

If balance is lost, quick step execution can prevent falls. Research has shown that speed of voluntary stepping was able to predict future falls in old adults. The aim of the study was to investigate voluntary stepping behavior, as well as to compare timing and leg push-off force-time relation parameters of involved and uninvolved legs in stroke survivors during single- and dual-task conditions. We also aimed to compare timing and leg push-off force-time relation parameters between stroke survivors and healthy individuals in both task conditions. Ten stroke survivors performed a voluntary step execution test with their involved and uninvolved legs under two conditions: while focusing only on the stepping task and while a separate attention-demanding task was performed simultaneously. Temporal parameters related to the step time were measured including the duration of the step initiation phase, the preparatory phase, the swing phase, and the total step time. In addition, force-time parameters representing the push-off power during stepping were calculated from ground reaction data and compared with 10 healthy controls. The involved legs of stroke survivors had a significantly slower stepping time than uninvolved legs due to increased swing phase duration during both single- and dual-task conditions. For dual compared to single task, the stepping time increased significantly due to a significant increase in the duration of step initiation. In general, the force time parameters were significantly different in both legs of stroke survivors as compared to healthy controls, with no significant effect of dual compared with single-task conditions in both groups. The inability of stroke survivors to swing the involved leg quickly may be the most significant factor contributing to the large number of falls to the paretic side. The results suggest that stroke survivors were unable to rapidly produce muscle force in fast actions. This may be the mechanism of delayed execution of a fast step when balance is lost, thus increasing the likelihood of falls in stroke survivors. Copyright © 2010 Elsevier Ltd. All rights reserved.

Phonics Pathways: Clear Steps to Easy Reading and Perfect Spelling. 10th Edition

ERIC Educational Resources Information Center

Hiskes, Dolores G.

2011-01-01

This tenth edition of the best-selling book teaches reading using sounds and spelling patterns. These sounds and patterns are introduced one at a time, and slowly built into words, syllables, phrases, and sentences. Simple step-by-step directions begin every lesson. Although originally designed for K-2 emergent readers, this award-winning book is…

"Cast Your Net Widely": Three Steps to Expanding and Refining Your Problem before Action Learning Application

ERIC Educational Resources Information Center

Reese, Simon R.

2015-01-01

This paper reflects upon a three-step process to expand the problem definition in the early stages of an action learning project. The process created a community-powered problem-solving approach within the action learning context. The simple three steps expanded upon in the paper create independence, dependence, and inter-dependence to aid the…

One-step detection of microRNA with high sensitivity and specificity via target-triggered loop-mediated isothermal amplification (TT-LAMP).

PubMed

Sun, Yuanyuan; Tian, Hui; Liu, Chenghui; Sun, Yueying; Li, Zhengping

2017-10-05

A novel one-step microRNA assay is developed based on a target-triggered loop-mediated isothermal amplification (TT-LAMP) mechanism, which enables the accurate detection of as low as 100 aM (1 zmol) microRNA with simple one-step operation by using only one-type of DNA polymerase.

A rapid and sensitive LC-MS/MS assay for the determination of saxagliptin and its active metabolite 5-hydroxy saxagliptin in human plasma and its application to a pharmacokinetic study.

PubMed

Batta, N; Pilli, N R; Derangula, V R; Vurimindi, H B; Damaramadugu, R; Yejella, R P

2015-03-01

The authors proposed a simple, rapid and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) assay method for the simultaneous determination of saxagliptin and its active metabolite 5-hydroxy saxagliptin in human plasma. The developed method was fully validated as per the US FDA guidelines. The method utilized stable labeled isotopes saxagliptin-15 N d2 (IS1) and 5-hydroxy saxagliptin-15 N-d2 (IS2) as internal standards for the quantification of saxagliptin and 5-hydroxy saxagliptin, respectively. Analytes and the internal standards were extracted from human plasma by a single step solid-phase extraction technique without drying, evaporation and reconstitution steps. The optimized mobile phase was composed of 0.1% acetic acid in 5 mM ammonium acetate and acetonitrile (30:70, v/v) and delivered at a flow rate of 0.85 mL/min. The method exhibits the linear calibration range of 0.05-100 ng/mL for both the analytes. The precision and accuracy results for both the analytes were well within the acceptance limits. The different stability experiments conducted in aqueous samples and in matrix samples are meeting the acceptance criteria. The chromatographic run time was set at 1.8 min; hence more than 400 samples can be analyzed in a single day. © Georg Thieme Verlag KG Stuttgart · New York.

Two-dimensional simulation and modeling in scanning electron microscope imaging and metrology research.

PubMed

Postek, Michael T; Vladár, András E; Lowney, Jeremiah R; Keery, William J

2002-01-01

Traditional Monte Carlo modeling of the electron beam-specimen interactions in a scanning electron microscope (SEM) produces information about electron beam penetration and output signal generation at either a single beam-landing location, or multiple landing positions. If the multiple landings lie on a line, the results can be graphed in a line scan-like format. Monte Carlo results formatted as line scans have proven useful in providing one-dimensional information about the sample (e.g., linewidth). When used this way, this process is called forward line scan modeling. In the present work, the concept of image simulation (or the first step in the inverse modeling of images) is introduced where the forward-modeled line scan data are carried one step further to construct theoretical two-dimensional (2-D) micrographs (i.e., theoretical SEM images) for comparison with similar experimentally obtained micrographs. This provides an ability to mimic and closely match theory and experiment using SEM images. Calculated and/or measured libraries of simulated images can be developed with this technique. The library concept will prove to be very useful in the determination of dimensional and other properties of simple structures, such as integrated circuit parts, where the shape of the features is preferably measured from a single top-down image or a line scan. This paper presents one approach to the generation of 2-D simulated images and presents some suggestions as to their application to critical dimension metrology.

A simple model of hysteresis behavior using spreadsheet analysis

NASA Astrophysics Data System (ADS)

Ehrmann, A.; Blachowicz, T.

2015-01-01

Hysteresis loops occur in many scientific and technical problems, especially as field dependent magnetization of ferromagnetic materials, but also as stress-strain-curves of materials measured by tensile tests including thermal effects, liquid-solid phase transitions, in cell biology or economics. While several mathematical models exist which aim to calculate hysteresis energies and other parameters, here we offer a simple model for a general hysteretic system, showing different hysteresis loops depending on the defined parameters. The calculation which is based on basic spreadsheet analysis plus an easy macro code can be used by students to understand how these systems work and how the parameters influence the reactions of the system on an external field. Importantly, in the step-by-step mode, each change of the system state, compared to the last step, becomes visible. The simple program can be developed further by several changes and additions, enabling the building of a tool which is capable of answering real physical questions in the broad field of magnetism as well as in other scientific areas, in which similar hysteresis loops occur.

Analysis of bacterial migration. 2: Studies with multiple attractant gradients

DOE Office of Scientific and Technical Information (OSTI.GOV)

Strauss, I.; Frymier, P.D.; Hahn, C.M.

1995-02-01

Many motile bacteria exhibit chemotaxis, the ability to bias their random motion toward or away from increasing concentrations of chemical substances which benefit or inhibit their survival, respectively. Since bacteria encounter numerous chemical concentration gradients simultaneously in natural surroundings, it is necessary to know quantitatively how a bacterial population responds in the presence of more than one chemical stimulus to develop predictive mathematical models describing bacterial migration in natural systems. This work evaluates three hypothetical models describing the integration of chemical signals from multiple stimuli: high sensitivity, maximum signal, and simple additivity. An expression for the tumbling probability for individualmore » stimuli is modified according to the proposed models and incorporated into the cell balance equation for a 1-D attractant gradient. Random motility and chemotactic sensitivity coefficients, required input parameters for the model, are measured for single stimulus responses. Theoretical predictions with the three signal integration models are compared to the net chemotactic response of Escherichia coli to co- and antidirectional gradients of D-fucose and [alpha]-methylaspartate in the stopped-flow diffusion chamber assay. Results eliminate the high-sensitivity model and favor the simple additivity over the maximum signal. None of the simple models, however, accurately predict the observed behavior, suggesting a more complex model with more steps in the signal processing mechanism is required to predict responses to multiple stimuli.« less

Effects of dual task on turning ability in stroke survivors and older adults.

PubMed

Hollands, K L; Agnihotri, D; Tyson, S F

2014-09-01

Turning is an integral component of independent mobility in which stroke survivors frequently fall. This study sought to measure the effects of competing cognitive demands on the stepping patterns of stroke survivors, compared to healthy age-match adults, during turning as a putative mechanism for falls. Walking and turning (90°) was assessed under single (walking and turning alone) and dual task (subtracting serial 3s while walking and turning) conditions using an electronic, pressure-sensitive walkway. Dependent measures were time to turn, variability in time to turn, step length, step width and single support time during three steps of the turn. Turning ability in single and dual task conditions was compared between stroke survivors (n=17, mean ± SD: 59 ± 113 months post-stroke, 64 ± 10 years of age) and age-matched healthy counterparts (n=15). Both groups took longer, were more variable, tended to widen the second step and, crucially, increased single support time on the inside leg of the turn while turning and distracted. Increased single support time during turning may represent biomechanical mechanism, within stepping patterns of turning under distraction, for increased risk of falls for both stroke survivors and older adults. Crown Copyright © 2014. Published by Elsevier B.V. All rights reserved.

DNA: Polymer and molecular code

NASA Astrophysics Data System (ADS)

Shivashankar, G. V.

1999-10-01

The thesis work focusses upon two aspects of DNA, the polymer and the molecular code. Our approach was to bring single molecule micromanipulation methods to the study of DNA. It included a home built optical microscope combined with an atomic force microscope and an optical tweezer. This combined approach led to a novel method to graft a single DNA molecule onto a force cantilever using the optical tweezer and local heating. With this method, a force versus extension assay of double stranded DNA was realized. The resolution was about 10 picoN. To improve on this force measurement resolution, a simple light backscattering technique was developed and used to probe the DNA polymer flexibility and its fluctuations. It combined the optical tweezer to trap a DNA tethered bead and the laser backscattering to detect the beads Brownian fluctuations. With this technique the resolution was about 0.1 picoN with a millisecond access time, and the whole entropic part of the DNA force-extension was measured. With this experimental strategy, we measured the polymerization of the protein RecA on an isolated double stranded DNA. We observed the progressive decoration of RecA on the l DNA molecule, which results in the extension of l , due to unwinding of the double helix. The dynamics of polymerization, the resulting change in the DNA entropic elasticity and the role of ATP hydrolysis were the main parts of the study. A simple model for RecA assembly on DNA was proposed. This work presents a first step in the study of genetic recombination. Recently we have started a study of equilibrium binding which utilizes fluorescence polarization methods to probe the polymerization of RecA on single stranded DNA. In addition to the study of material properties of DNA and DNA-RecA, we have developed experiments for which the code of the DNA is central. We studied one aspect of DNA as a molecular code, using different techniques. In particular the programmatic use of template specificity makes gene expression a prime example of a biological code. We developed a novel method of making DNA micro- arrays, the so-called DNA chip. Using the optical tweezer concept, we were able to pattern biomolecules on a solid substrate, developing a new type of sub-micron laser lithography. A laser beam is focused onto a thin gold film on a glass substrate. Laser ablation of gold results in local aggregation of nanometer scale beads conjugated with small DNA oligonucleotides, with sub-micron resolution. This leads to specific detection of cDNA and RNA molecules. We built a simple micro-array fabrication and detection in the laboratory, based on this method, to probe addressable pools (genes, proteins or antibodies). We have lately used molecular beacons (single stranded DNA with a stem-loop structure containing a fluorophore and quencher), for the direct detection of unlabelled mRNA. As a first step towards a study of the dynamics of the biological code, we have begun to examine the patterns of gene expression during virus (T7 phage) infection of E-coli bacteria.

Trunk-acceleration based assessment of gait parameters in older persons: a comparison of reliability and validity of four inverted pendulum based estimations.

PubMed

Zijlstra, Agnes; Zijlstra, Wiebren

2013-09-01

Inverted pendulum (IP) models of human walking allow for wearable motion-sensor based estimations of spatio-temporal gait parameters during unconstrained walking in daily-life conditions. At present it is unclear to what extent different IP based estimations yield different results, and reliability and validity have not been investigated in older persons without a specific medical condition. The aim of this study was to compare reliability and validity of four different IP based estimations of mean step length in independent-living older persons. Participants were assessed twice and walked at different speeds while wearing a tri-axial accelerometer at the lower back. For all step-length estimators, test-retest intra-class correlations approached or were above 0.90. Intra-class correlations with reference step length were above 0.92 with a mean error of 0.0 cm when (1) multiplying the estimated center-of-mass displacement during a step by an individual correction factor in a simple IP model, or (2) adding an individual constant for bipedal stance displacement to the estimated displacement during single stance in a 2-phase IP model. When applying generic corrections or constants in all subjects (i.e. multiplication by 1.25, or adding 75% of foot length), correlations were above 0.75 with a mean error of respectively 2.0 and 1.2 cm. Although the results indicate that an individual adjustment of the IP models provides better estimations of mean step length, the ease of a generic adjustment can be favored when merely evaluating intra-individual differences. Further studies should determine the validity of these IP based estimations for assessing gait in daily life. Copyright © 2013 Elsevier B.V. All rights reserved.

Regression Analysis of a Disease Onset Distribution Using Diagnosis Data

PubMed Central

Young, Jessica G.; Jewell, Nicholas P.; Samuels, Steven J.

2008-01-01

Summary We consider methods for estimating the effect of a covariate on a disease onset distribution when the observed data structure consists of right-censored data on diagnosis times and current status data on onset times amongst individuals who have not yet been diagnosed. Dunson and Baird (2001, Biometrics 57, 306–403) approached this problem using maximum likelihood, under the assumption that the ratio of the diagnosis and onset distributions is monotonic nondecreasing. As an alternative, we propose a two-step estimator, an extension of the approach of van der Laan, Jewell, and Petersen (1997, Biometrika 84, 539–554) in the single sample setting, which is computationally much simpler and requires no assumptions on this ratio. A simulation study is performed comparing estimates obtained from these two approaches, as well as that from a standard current status analysis that ignores diagnosis data. Results indicate that the Dunson and Baird estimator outperforms the two-step estimator when the monotonicity assumption holds, but the reverse is true when the assumption fails. The simple current status estimator loses only a small amount of precision in comparison to the two-step procedure but requires monitoring time information for all individuals. In the data that motivated this work, a study of uterine fibroids and chemical exposure to dioxin, the monotonicity assumption is seen to fail. Here, the two-step and current status estimators both show no significant association between the level of dioxin exposure and the hazard for onset of uterine fibroids; the two-step estimator of the relative hazard associated with increasing levels of exposure has the least estimated variance amongst the three estimators considered. PMID:17680832

Outcome of vesicourethral anastomosis after robot-assisted laparoscopic radical prostatectomy: A 6-year experience in Taiwan.

PubMed

Chen, Cheng-Che; Yang, Cheng-Kuang; Hung, Siu-Wan; Wang, John; Ou, Yen-Chuan

2015-10-01

The use of a da Vinci robotic system may improve the outcome of urological surgery. This study reports 6 years of experience with vesicourethral anastomosis (VUA) following robot-assisted laparoscopic radical prostatectomy (RALP) performed in Taichung Veterans General Hospital, Taichung, Taiwan. A total of 350 patients who underwent RALP by a single surgeon were reviewed. We followed Dr Patel's RALP procedure with minor modifications. VUA was checked with 120 mL and 200 mL saline in sequence. The urinary bladder was then pressed with endoscopic instruments. If a VUA leak was detected, it was sutured immediately. An 18-French silicon Foley's catheter was inserted and removed 7-14 days after RALP. Preoperative characteristics and perioperative complications were assessed. Overall, 332 (94.85%) patients were without any leakage in the first step of the challenge, eight of whom had leakage in the second step. After repair, all were free from leakage. The other 18 patients had leakage in the first step of the challenge (5.14%). After repair, 12 patients were without leakage in the second step. However, one patient had urine leakage postoperatively. The other six patients had leakage in the second step. After repair, two patients were free from leakage, but the remaining four suffered from persistent minor urine leakage postoperatively. The urine leakage rate after RALP was 1.43% (5/350). The potential urine leakage after bladder challenge and endoscopic instruments pressing could be minimized to 0.29% (1/346). VUA leakage after RALP is rare. Intraoperative VUA challenge is simple and feasible compared to postoperative retrograde cystography. Copyright © 2014. Published by Elsevier B.V.

Single Spore Isolation as a Simple and Efficient Technique to obtain fungal pure culture

NASA Astrophysics Data System (ADS)

Noman, E.; Al-Gheethi, AA; Rahman, N. K.; Talip, B.; Mohamed, R.; H, N.; Kadir, O. A.

2018-04-01

The successful identification of fungi by phenotypic methods or molecular technique depends mainly on the using an advanced technique for purifying the isolates. The most efficient is the single spore technique due to the simple requirements and the efficiency in preventing the contamination by yeast, mites or bacteria. The method described in the present work is depends on the using of a light microscope to transfer one spore into a new culture medium. The present work describes a simple and efficient procedure for single spore isolation to purify of fungi recovered from the clinical wastes.

Differentiation of Mycobacterium tuberculosis complex from non-tubercular mycobacteria by nested multiplex PCR targeting IS6110, MTP40 and 32kD alpha antigen encoding gene fragments.

PubMed

Sinha, Pallavi; Gupta, Anamika; Prakash, Pradyot; Anupurba, Shampa; Tripathi, Rajneesh; Srivastava, G N

2016-03-12

Control of the global burden of tuberculosis is obstructed due to lack of simple, rapid and cost effective diagnostic techniques that can be used in resource poor-settings. To facilitate the early diagnosis of TB directly from clinical specimens, we have standardized and validated the use of nested multiplex PCR, targeting gene fragments IS6110, MTP40 and 32kD α-antigen encoding genes specific for Mycobacterium tuberculosis complex and non-tubercular mycobacteria (NTM), in comparison to smear microscopy, solid culture and single step multiplex PCR. The results were evaluated in comparison to a composite reference standard (CRS) comprising of microbiological results (smear and culture), clinical, radiological and cytopathological findings, clinical treatment and response to anti-tubercular therapy. The nested multiplex PCR (nMPCR) assay was evaluated to test its utility in 600 (535 pulmonary and 65 extra-pulmonary specimens) clinically suspected TB cases. All specimens were processed for smear, culture, single step multiplex PCR and nested multiplex PCR testing. Out of 535 screened pulmonary and 65 extra-pulmonary specimens, 329 (61.5%) and 19 (29.2%) cases were culture positive for M. tuberculosis. Based on CRS, 450 patients had "clinical TB" (definitive-TB, probable-TB and possible-TB). Remaining 150 were confirmed "non-TB" cases. For culture, the sensitivity was low, 79.3% for pulmonary and 54.3% for extra-pulmonary cases. The sensitivity and specificity results for nMPCR test were evaluated taken composite reference standard as a gold standard. The sensitivity of the nMPCR assay was 97.1% for pulmonary and 91.4% for extra-pulmonary TB cases with specificity of 100% and 93.3% respectively. Nested multiplex PCR using three gene primers is a rapid, reliable and highly sensitive and specific diagnostic technique for the detection and differentiation of M. tuberculosis complex from NTM genome and will be useful in diagnosing paucibacillary samples. Nested multiplex PCR assay was found to be better than single step multiplex PCR for assessing the diagnosis of TB.

Argon behaviour in an inverted Barrovian sequence, Sikkim Himalaya: The consequences of temperature and timescale on 40Ar/39Ar mica geochronology

NASA Astrophysics Data System (ADS)

Mottram, Catherine M.; Warren, Clare J.; Halton, Alison M.; Kelley, Simon P.; Harris, Nigel B. W.

2015-12-01

40Ar/39Ar dating of metamorphic rocks sometimes yields complicated datasets which are difficult to interpret in terms of timescales of the metamorphic cycle. Single-grain fusion and step-heating data were obtained for rocks sampled through a major thrust-sense shear zone (the Main Central Thrust) and the associated inverted metamorphic zone in the Sikkim region of the eastern Himalaya. This transect provides a natural laboratory to explore factors influencing apparent 40Ar/39Ar ages in similar lithologies at a variety of metamorphic pressure and temperature (P-T) conditions. The 40Ar/39Ar dataset records progressively younger apparent age populations and a decrease in within-sample dispersion with increasing temperature through the sequence. The white mica populations span 2-9 Ma within each sample in the structurally lower levels (garnet grade) but only 0-3 Ma at structurally higher levels (kyanite-sillimanite grade). Mean white mica single-grain fusion population ages vary from 16.2 ± 3.9 Ma (2σ) to 13.2 ± 1.3 Ma (2σ) from lowest to highest levels. White mica step-heating data from the same samples yields plateau ages from 14.27 ± 0.13 Ma to 12.96 ± 0.05 Ma. Biotite yield older apparent age populations with mean single-grain fusion dates varying from 74.7 ± 11.8 Ma (2σ) at the lowest structural levels to 18.6 ± 4.7 Ma (2σ) at the highest structural levels; the step-heating plateaux are commonly disturbed. Temperatures > 600 °C at pressures of 0.4-0.8 GPa sustained over > 5 Ma, appear to be required for white mica and biotite ages to be consistent with diffusive, open-system cooling. At lower temperatures, and/or over shorter metamorphic timescales, more 40Ar is retained than results from simple diffusion models suggest. Diffusion modelling of Ar in white mica from the highest structural levels suggests that the high-temperature rocks cooled at a rate of 50-80 °C Ma- 1, consistent with rapid thrusting, extrusion and exhumation along the Main Central Thrust during the mid-Miocene.

Auxotonic to isometric contraction transitioning in a beating heart causes myosin step-size to down shift

PubMed Central

Sun, Xiaojing; Wang, Yihua; Ajtai, Katalin

2017-01-01

Myosin motors in cardiac ventriculum convert ATP free energy to the work of moving blood volume under pressure. The actin bound motor cyclically rotates its lever-arm/light-chain complex linking motor generated torque to the myosin filament backbone and translating actin against resisting force. Previous research showed that the unloaded in vitro motor is described with high precision by single molecule mechanical characteristics including unitary step-sizes of approximately 3, 5, and 8 nm and their relative step-frequencies of approximately 13, 50, and 37%. The 3 and 8 nm unitary step-sizes are dependent on myosin essential light chain (ELC) N-terminus actin binding. Step-size and step-frequency quantitation specifies in vitro motor function including duty-ratio, power, and strain sensitivity metrics. In vivo, motors integrated into the muscle sarcomere form the more complex and hierarchically functioning muscle machine. The goal of the research reported here is to measure single myosin step-size and step-frequency in vivo to assess how tissue integration impacts motor function. A photoactivatable GFP tags the ventriculum myosin lever-arm/light-chain complex in the beating heart of a live zebrafish embryo. Detected single GFP emission reports time-resolved myosin lever-arm orientation interpreted as step-size and step-frequency providing single myosin mechanical characteristics over the active cycle. Following step-frequency of cardiac ventriculum myosin transitioning from low to high force in relaxed to auxotonic to isometric contraction phases indicates that the imposition of resisting force during contraction causes the motor to down-shift to the 3 nm step-size accounting for >80% of all the steps in the near-isometric phase. At peak force, the ATP initiated actomyosin dissociation is the predominant strain inhibited transition in the native myosin contraction cycle. The proposed model for motor down-shifting and strain sensing involves ELC N-terminus actin binding. Overall, the approach is a unique bottom-up single molecule mechanical characterization of a hierarchically functional native muscle myosin. PMID:28423017

Comparison of Stepped Care Delivery Against a Single, Empirically Validated Cognitive-Behavioral Therapy Program for Youth With Anxiety: A Randomized Clinical Trial.

PubMed

Rapee, Ronald M; Lyneham, Heidi J; Wuthrich, Viviana; Chatterton, Mary Lou; Hudson, Jennifer L; Kangas, Maria; Mihalopoulos, Cathrine

2017-10-01

Stepped care is embraced as an ideal model of service delivery but is minimally evaluated. The aim of this study was to evaluate the efficacy of cognitive-behavioral therapy (CBT) for child anxiety delivered via a stepped-care framework compared against a single, empirically validated program. A total of 281 youth with anxiety disorders (6-17 years of age) were randomly allocated to receive either empirically validated treatment or stepped care involving the following: (1) low intensity; (2) standard CBT; and (3) individually tailored treatment. Therapist qualifications increased at each step. Interventions did not differ significantly on any outcome measures. Total therapist time per child was significantly shorter to deliver stepped care (774 minutes) compared with best practice (897 minutes). Within stepped care, the first 2 steps returned the strongest treatment gains. Stepped care and a single empirically validated program for youth with anxiety produced similar efficacy, but stepped care required slightly less therapist time. Restricting stepped care to only steps 1 and 2 would have led to considerable time saving with modest loss in efficacy. Clinical trial registration information-A Randomised Controlled Trial of Standard Care Versus Stepped Care for Children and Adolescents With Anxiety Disorders; http://anzctr.org.au/; ACTRN12612000351819. Copyright © 2017 American Academy of Child and Adolescent Psychiatry. Published by Elsevier Inc. All rights reserved.

Step Forward. Single Parent/Homemaker Annual Report for the Fiscal Year 1990-1991.

ERIC Educational Resources Information Center

Kentucky Tech Region 5, Elizabethtown.

The Step Forward Single Parent/Homemaker Program in Elizabethtown, Kentucky, was developed to provide information on career opportunities and assist the target individuals in career assessment, career counseling, and goal setting in order to develop self-esteem and time management skills. During the second year of the Step Forward program in…

Correlation among electronegativity, cation polarizability, optical basicity and single bond strength of simple oxides

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dimitrov, Vesselin, E-mail: vesselin@uctm.edu; Komatsu, Takayuki, E-mail: komatsu@mst.nagaokaut.ac.jp

2012-12-15

A suitable relationship between free-cation polarizability and electronegativity of elements in different valence states and with the most common coordination numbers has been searched on the basis of the similarity in physical nature of both quantities. In general, the cation polarizability increases with decreasing element electronegativity. A systematic periodic change in the polarizability against the electronegativity has been observed in the isoelectronic series. It has been found that generally the optical basicity increases and the single bond strength of simple oxides decreases with decreasing the electronegativity. The observed trends have been discussed on the basis of electron donation ability ofmore » the oxide ions and type of chemical bonding in simple oxides. - Graphical abstract: This figure shows the single bond strength of simple oxides as a function of element electronegativity. A remarkable correlation exists between these independently obtained quantities. High values of electronegativity correspond to high values of single bond strength and vice versa. It is obvious that the observed trend in this figure is closely related to the type of chemical bonding in corresponding oxide. Highlights: Black-Right-Pointing-Pointer A suitable relationship between free-cation polarizability and electronegativity of elements was searched. Black-Right-Pointing-Pointer The cation polarizability increases with decreasing element electronegativity. Black-Right-Pointing-Pointer The single bond strength of simple oxides decreases with decreasing the electronegativity. Black-Right-Pointing-Pointer The observed trends were discussed on the basis of type of chemical bonding in simple oxides.« less

Loop-mediated isothermal amplification assay for detection and discrimination of Toxocara canis and Toxocara cati eggs directly from sand samples.

PubMed

Macuhova, K; Kumagai, T; Akao, N; Ohta, N

2010-12-01

We developed a novel and simple method, using loop-mediated isothermal amplification (LAMP), for the detection and discrimination of Toxocara canis and Toxocara cati eggs. The new method employs 4 steps: (1) concentration of Toxocara eggs in a small amount of sand; (2) dissolution of the proteinaceous membrane of eggs and simultaneously separation of them from the sand using NaClO treatment; (3) extraction of DNA using NaOH treatment; and (4) detection of T. canis / T. cati DNA using a LAMP assay. All these steps are fast, easy to perform, and do not require expensive equipment or reagents. The novel method was tested both experimentally and in a field study. In the laboratory, we could reliably detect as few as 3 T. canis eggs in artificially contaminated sand, if the experiment was repeated twice. In the field trial, we were able to detect T. cati DNA from 4 natural sandpits having moderate to heavy contamination, although not in a single lightly contaminated sandpit. All of the examined sandpits were found to be contaminated with eggs of T. cati, but none appeared to contain T. canis. Our new method could extract DNA from T. canis and T. cati eggs directly from sand samples as well as detect and distinguish these 2 species in a few easy steps, with markedly reduced time and expense.

Disordered Nanohole Patterns in Metal-Insulator Multilayer for Ultra-broadband Light Absorption: Atomic Layer Deposition for Lithography Free Highly repeatable Large Scale Multilayer Growth.

PubMed

Ghobadi, Amir; Hajian, Hodjat; Dereshgi, Sina Abedini; Bozok, Berkay; Butun, Bayram; Ozbay, Ekmel

2017-11-08

In this paper, we demonstrate a facile, lithography free, and large scale compatible fabrication route to synthesize an ultra-broadband wide angle perfect absorber based on metal-insulator-metal-insulator (MIMI) stack design. We first conduct a simulation and theoretical modeling approach to study the impact of different geometries in overall stack absorption. Then, a Pt-Al 2 O 3 multilayer is fabricated using a single atomic layer deposition (ALD) step that offers high repeatability and simplicity in the fabrication step. In the best case, we get an absorption bandwidth (BW) of 600 nm covering a range of 400 nm-1000 nm. A substantial improvement in the absorption BW is attained by incorporating a plasmonic design into the middle Pt layer. Our characterization results demonstrate that the best configuration can have absorption over 0.9 covering a wavelength span of 400 nm-1490 nm with a BW that is 1.8 times broader compared to that of planar design. On the other side, the proposed structure retains its absorption high at angles as wide as 70°. The results presented here can serve as a beacon for future performance enhanced multilayer designs where a simple fabrication step can boost the overall device response without changing its overall thickness and fabrication simplicity.

One-step sol-gel imprint lithography for guided-mode resonance structures.

PubMed

Huang, Yin; Liu, Longju; Johnson, Michael; C Hillier, Andrew; Lu, Meng

2016-03-04

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.

A new method to characterize the kinetics of cholinesterases inhibited by carbamates.

PubMed

Xiao, Qiaoling; Zhou, Huimin; Wei, Hong; Du, Huaqiao; Tan, Wen; Zhan, Yiyi; Pistolozzi, Marco

2017-09-10

The inhibition of cholinesterases (ChEs) by carbamates includes a carbamylation (inhibition) step, in which the drug transfers its carbamate moiety to the active site of the enzyme and a decarbamylation (activity recovery) step, in which the carbamyl group is hydrolyzed from the enzyme. The carbamylation and decarbamylation kinetics decide the extent and the duration of the inhibition, thus the full characterization of candidate carbamate inhibitors requires the measurement of the kinetic constants describing both steps. Carbamylation and decarbamylation rate constants are traditionally measured by two separate set of experiments, thus making the full characterization of candidate inhibitors time-consuming. In this communication we show that by the analysis of the area under the inhibition-time curve of cholinesterases inhibited by carbamates it is possible to calculate the decarbamylation rate constant from the same data traditionally used to characterize only the carbamylation kinetics, therefore it is possible to obtain a full characterization of the inhibition with a single set of experiments. The characterization of the inhibition kinetics of human and dog plasma butyrylcholinesterase and of human acetylcholinesterase by bambuterol and bambuterol monocarbamate enantiomers was used to demonstrate the validity of the approach. The results showed that the proposed method provides reliable estimations of carbamylation and decarbamylation rate constants thus representing a simple and useful approach to reduce the time required for the characterization of carbamate inhibitors. Copyright © 2017 Elsevier B.V. All rights reserved.

LACO-Wiki: A land cover validation tool and a new, innovative teaching resource for remote sensing and the geosciences

NASA Astrophysics Data System (ADS)

See, Linda; Perger, Christoph; Dresel, Christopher; Hofer, Martin; Weichselbaum, Juergen; Mondel, Thomas; Steffen, Fritz

2016-04-01

The validation of land cover products is an important step in the workflow of generating a land cover map from remotely-sensed imagery. Many students of remote sensing will be given exercises on classifying a land cover map followed by the validation process. Many algorithms exist for classification, embedded within proprietary image processing software or increasingly as open source tools. However, there is little standardization for land cover validation, nor a set of open tools available for implementing this process. The LACO-Wiki tool was developed as a way of filling this gap, bringing together standardized land cover validation methods and workflows into a single portal. This includes the storage and management of land cover maps and validation data; step-by-step instructions to guide users through the validation process; sound sampling designs; an easy-to-use environment for validation sample interpretation; and the generation of accuracy reports based on the validation process. The tool was developed for a range of users including producers of land cover maps, researchers, teachers and students. The use of such a tool could be embedded within the curriculum of remote sensing courses at a university level but is simple enough for use by students aged 13-18. A beta version of the tool is available for testing at: http://www.laco-wiki.net.

A Step-by-Step Picture of Pulsed (Time-Domain) NMR.

ERIC Educational Resources Information Center

Schwartz, Leslie J.

1988-01-01

Discusses a method for teaching time pulsed NMR principals that are as simple and pictorial as possible. Uses xyz coordinate figures and presents theoretical explanations using a Fourier transformation spectrum. Assumes no previous knowledge of quantum mechanics for students. Usable for undergraduates. (MVL)

Heat damaged forages: effects on forage energy content

USDA-ARS?s Scientific Manuscript database

Traditionally, educational materials describing the effects of heat damage within baled hays have focused on reduced bioavailability of crude protein as a result of Maillard reactions. These reactions are not simple, but actually occur in complex, multi-step pathways. Typically, the initial step inv...

Changes in step-width during dual-task walking predicts falls.

PubMed

Nordin, E; Moe-Nilssen, R; Ramnemark, A; Lundin-Olsson, L

2010-05-01

The aim was to evaluate whether gait pattern changes between single- and dual-task conditions were associated with risk of falling in older people. Dual-task cost (DTC) of 230 community living, physically independent people, 75 years or older, was determined with an electronic walkway. Participants were followed up each month for 1 year to record falls. Mean and variability measures of gait characteristics for 5 dual-task conditions were compared to single-task walking for each participant. Almost half (48%) of the participants fell at least once during follow-up. Risk of falling increased in individuals where DTC for performing a subtraction task demonstrated change in mean step-width compared to single-task walking. Risk of falling decreased in individuals where DTC for carrying a cup and saucer demonstrated change compared to single-task walking in mean step-width, mean step-time, and step-length variability. Degree of change in gait characteristics related to a change in risk of falling differed between measures. Prognostic guidance for fall risk was found for the above DTCs in mean step-width with a negative likelihood ratio of 0.5 and a positive likelihood ratio of 2.3, respectively. Findings suggest that changes in step-width, step-time, and step-length with dual tasking may be related to future risk of falling. Depending on the nature of the second task, DTC may indicate either an increased risk of falling, or a protective strategy to avoid falling. Copyright 2010. Published by Elsevier B.V.

A step-by-step guide to non-linear regression analysis of experimental data using a Microsoft Excel spreadsheet.

PubMed

Brown, A M

2001-06-01

The objective of this present study was to introduce a simple, easily understood method for carrying out non-linear regression analysis based on user input functions. While it is relatively straightforward to fit data with simple functions such as linear or logarithmic functions, fitting data with more complicated non-linear functions is more difficult. Commercial specialist programmes are available that will carry out this analysis, but these programmes are expensive and are not intuitive to learn. An alternative method described here is to use the SOLVER function of the ubiquitous spreadsheet programme Microsoft Excel, which employs an iterative least squares fitting routine to produce the optimal goodness of fit between data and function. The intent of this paper is to lead the reader through an easily understood step-by-step guide to implementing this method, which can be applied to any function in the form y=f(x), and is well suited to fast, reliable analysis of data in all fields of biology.

DOPI and PALM imaging of single carbohydrate binding modules bound to cellulose nanocrystals

NASA Astrophysics Data System (ADS)

Dagel, D. J.; Liu, Y.-S.; Zhong, L.; Luo, Y.; Zeng, Y.; Himmel, M.; Ding, S.-Y.; Smith, S.

2011-03-01

We use single molecule imaging methods to study the binding characteristics of carbohydrate-binding modules (CBMs) to cellulose crystals. The CBMs are carbohydrate specific binding proteins, and a functional component of most cellulase enzymes, which in turn hydrolyze cellulose, releasing simple sugars suitable for fermentation to biofuels. The CBM plays the important role of locating the crystalline face of cellulose, a critical step in cellulase action. A biophysical understanding of the CBM action aids in developing a mechanistic picture of the cellulase enzyme, important for selection and potential modification. Towards this end, we have genetically modified cellulose-binding CBM derived from bacterial source with green fluorescent protein (GFP), and photo-activated fluorescence protein PAmCherry tags, respectively. Using the single molecule method known as Defocused Orientation and Position Imaging (DOPI), we observe a preferred orientation of the CBM-GFP complex relative to the Valonia cellulose nanocrystals. Subsequent analysis showed the CBMs bind to the opposite hydrophobic <110> faces of the cellulose nanocrystals with a welldefined cross-orientation of about { 70°. Photo Activated Localization Microscopy (PALM) is used to localize CBMPAmCherry with a localization accuracy of { 10nm. Analysis of the nearest neighbor distributions along and perpendicular to the cellulose nanocrystal axes are consistent with single-file CBM binding along the fiber axis, and microfibril bundles consisting of close packed { 20nm or smaller cellulose microfibrils.

Method of upgrading oils containing hydroxyaromatic hydrocarbon compounds to highly aromatic gasoline

DOEpatents

Baker, Eddie G.; Elliott, Douglas C.

1993-01-01

The present invention is a multi-stepped method of converting an oil which is produced by various biomass and coal conversion processes and contains primarily single and multiple ring hydroxyaromatic hydrocarbon compounds to highly aromatic gasoline. The single and multiple ring hydroxyaromatic hydrocarbon compounds in a raw oil material are first deoxygenated to produce a deoxygenated oil material containing single and multiple ring aromatic compounds. Then, water is removed from the deoxygenated oil material. The next step is distillation to remove the single ring aromatic compouns as gasoline. In the third step, the multiple ring aromatics remaining in the deoxygenated oil material are cracked in the presence of hydrogen to produce a cracked oil material containing single ring aromatic compounds. Finally, the cracked oil material is then distilled to remove the single ring aromatics as gasoline.

Method of upgrading oils containing hydroxyaromatic hydrocarbon compounds to highly aromatic gasoline

DOEpatents

Baker, E.G.; Elliott, D.C.

1993-01-19

The present invention is a multi-stepped method of converting an oil which is produced by various biomass and coal conversion processes and contains primarily single and multiple ring hydroxyaromatic hydrocarbon compounds to highly aromatic gasoline. The single and multiple ring hydroxyaromatic hydrocarbon compounds in a raw oil material are first deoxygenated to produce a deoxygenated oil material containing single and multiple ring aromatic compounds. Then, water is removed from the deoxygenated oil material. The next step is distillation to remove the single ring aromatic compounds as gasoline. In the third step, the multiple ring aromatics remaining in the deoxygenated oil material are cracked in the presence of hydrogen to produce a cracked oil material containing single ring aromatic compounds. Finally, the cracked oil material is then distilled to remove the single ring aromatics as gasoline.

Single molecule thermodynamics in biological motors.

PubMed

Taniguchi, Yuichi; Karagiannis, Peter; Nishiyama, Masayoshi; Ishii, Yoshiharu; Yanagida, Toshio

2007-04-01

Biological molecular machines use thermal activation energy to carry out various functions. The process of thermal activation has the stochastic nature of output events that can be described according to the laws of thermodynamics. Recently developed single molecule detection techniques have allowed each distinct enzymatic event of single biological machines to be characterized providing clues to the underlying thermodynamics. In this study, the thermodynamic properties in the stepping movement of a biological molecular motor have been examined. A single molecule detection technique was used to measure the stepping movements at various loads and temperatures and a range of thermodynamic parameters associated with the production of each forward and backward step including free energy, enthalpy, entropy and characteristic distance were obtained. The results show that an asymmetry in entropy is a primary factor that controls the direction in which the motor will step. The investigation on single molecule thermodynamics has the potential to reveal dynamic properties underlying the mechanisms of how biological molecular machines work.

Cell fixation and preservation for droplet-based single-cell transcriptomics.

PubMed

Alles, Jonathan; Karaiskos, Nikos; Praktiknjo, Samantha D; Grosswendt, Stefanie; Wahle, Philipp; Ruffault, Pierre-Louis; Ayoub, Salah; Schreyer, Luisa; Boltengagen, Anastasiya; Birchmeier, Carmen; Zinzen, Robert; Kocks, Christine; Rajewsky, Nikolaus

2017-05-19

Recent developments in droplet-based microfluidics allow the transcriptional profiling of thousands of individual cells in a quantitative, highly parallel and cost-effective way. A critical, often limiting step is the preparation of cells in an unperturbed state, not altered by stress or ageing. Other challenges are rare cells that need to be collected over several days or samples prepared at different times or locations. Here, we used chemical fixation to address these problems. Methanol fixation allowed us to stabilise and preserve dissociated cells for weeks without compromising single-cell RNA sequencing data. By using mixtures of fixed, cultured human and mouse cells, we first showed that individual transcriptomes could be confidently assigned to one of the two species. Single-cell gene expression from live and fixed samples correlated well with bulk mRNA-seq data. We then applied methanol fixation to transcriptionally profile primary cells from dissociated, complex tissues. Low RNA content cells from Drosophila embryos, as well as mouse hindbrain and cerebellum cells prepared by fluorescence-activated cell sorting, were successfully analysed after fixation, storage and single-cell droplet RNA-seq. We were able to identify diverse cell populations, including neuronal subtypes. As an additional resource, we provide 'dropbead', an R package for exploratory data analysis, visualization and filtering of Drop-seq data. We expect that the availability of a simple cell fixation method will open up many new opportunities in diverse biological contexts to analyse transcriptional dynamics at single-cell resolution.

Transcript-specific, single-nucleotide polymorphism discovery and linkage analysis in hexaploid bread wheat (Triticum aestivum L.).

PubMed

Allen, Alexandra M; Barker, Gary L A; Berry, Simon T; Coghill, Jane A; Gwilliam, Rhian; Kirby, Susan; Robinson, Phil; Brenchley, Rachel C; D'Amore, Rosalinda; McKenzie, Neil; Waite, Darren; Hall, Anthony; Bevan, Michael; Hall, Neil; Edwards, Keith J

2011-12-01

Food security is a global concern and substantial yield increases in cereal crops are required to feed the growing world population. Wheat is one of the three most important crops for human and livestock feed. However, the complexity of the genome coupled with a decline in genetic diversity within modern elite cultivars has hindered the application of marker-assisted selection (MAS) in breeding programmes. A crucial step in the successful application of MAS in breeding programmes is the development of cheap and easy to use molecular markers, such as single-nucleotide polymorphisms. To mine selected elite wheat germplasm for intervarietal single-nucleotide polymorphisms, we have used expressed sequence tags derived from public sequencing programmes and next-generation sequencing of normalized wheat complementary DNA libraries, in combination with a novel sequence alignment and assembly approach. Here, we describe the development and validation of a panel of 1114 single-nucleotide polymorphisms in hexaploid bread wheat using competitive allele-specific polymerase chain reaction genotyping technology. We report the genotyping results of these markers on 23 wheat varieties, selected to represent a broad cross-section of wheat germplasm including a number of elite UK varieties. Finally, we show that, using relatively simple technology, it is possible to rapidly generate a linkage map containing several hundred single-nucleotide polymorphism markers in the doubled haploid mapping population of Avalon × Cadenza. © 2011 The Authors. Plant Biotechnology Journal © 2011 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.

An efficient method for variable region assembly in the construction of scFv phage display libraries using independent strand amplification

PubMed Central

Sotelo, Pablo H.; Collazo, Noberto; Zuñiga, Roberto; Gutiérrez-González, Matías; Catalán, Diego; Ribeiro, Carolina Hager; Aguillón, Juan Carlos; Molina, María Carmen

2012-01-01

Phage display library technology is a common method to produce human antibodies. In this technique, the immunoglobulin variable regions are displayed in a bacteriophage in a way that each filamentous virus displays the product of a single antibody gene on its surface. From the collection of different phages, it is possible to isolate the virus that recognizes specific targets. The most common form in which to display antibody variable regions in the phage is the single chain variable fragment format (scFv), which requires assembly of the heavy and light immunoglobulin variable regions in a single gene. In this work, we describe a simple and efficient method for the assembly of immunoglobulin heavy and light chain variable regions in a scFv format. This procedure involves a two-step reaction: (1) DNA amplification to produce the single strand form of the heavy or light chain gene required for the fusion; and (2) mixture of both single strand products followed by an assembly reaction to construct a complete scFv gene. Using this method, we produced 6-fold more scFv encoding DNA than the commonly used splicing by overlap extension PCR (SOE-PCR) approach. The scFv gene produced by this method also proved to be efficient in generating a diverse scFv phage display library. From this scFv library, we obtained phages that bound several non-related antigens, including recombinant proteins and rotavirus particles. PMID:22692130

Genomic prediction in a nuclear population of layers using single-step models.

PubMed

Yan, Yiyuan; Wu, Guiqin; Liu, Aiqiao; Sun, Congjiao; Han, Wenpeng; Li, Guangqi; Yang, Ning

2018-02-01

Single-step genomic prediction method has been proposed to improve the accuracy of genomic prediction by incorporating information of both genotyped and ungenotyped animals. The objective of this study is to compare the prediction performance of single-step model with a 2-step models and the pedigree-based models in a nuclear population of layers. A total of 1,344 chickens across 4 generations were genotyped by a 600 K SNP chip. Four traits were analyzed, i.e., body weight at 28 wk (BW28), egg weight at 28 wk (EW28), laying rate at 38 wk (LR38), and Haugh unit at 36 wk (HU36). In predicting offsprings, individuals from generation 1 to 3 were used as training data and females from generation 4 were used as validation set. The accuracies of predicted breeding values by pedigree BLUP (PBLUP), genomic BLUP (GBLUP), SSGBLUP and single-step blending (SSBlending) were compared for both genotyped and ungenotyped individuals. For genotyped females, GBLUP performed no better than PBLUP because of the small size of training data, while the 2 single-step models predicted more accurately than the PBLUP model. The average predictive ability of SSGBLUP and SSBlending were 16.0% and 10.8% higher than the PBLUP model across traits, respectively. Furthermore, the predictive abilities for ungenotyped individuals were also enhanced. The average improvements of prediction abilities were 5.9% and 1.5% for SSGBLUP and SSBlending model, respectively. It was concluded that single-step models, especially the SSGBLUP model, can yield more accurate prediction of genetic merits and are preferable for practical implementation of genomic selection in layers. © 2017 Poultry Science Association Inc.

Multi-target-qubit unconventional geometric phase gate in a multi-cavity system

NASA Astrophysics Data System (ADS)

Liu, Tong; Cao, Xiao-Zhi; Su, Qi-Ping; Xiong, Shao-Jie; Yang, Chui-Ping

2016-02-01

Cavity-based large scale quantum information processing (QIP) may involve multiple cavities and require performing various quantum logic operations on qubits distributed in different cavities. Geometric-phase-based quantum computing has drawn much attention recently, which offers advantages against inaccuracies and local fluctuations. In addition, multiqubit gates are particularly appealing and play important roles in QIP. We here present a simple and efficient scheme for realizing a multi-target-qubit unconventional geometric phase gate in a multi-cavity system. This multiqubit phase gate has a common control qubit but different target qubits distributed in different cavities, which can be achieved using a single-step operation. The gate operation time is independent of the number of qubits and only two levels for each qubit are needed. This multiqubit gate is generic, e.g., by performing single-qubit operations, it can be converted into two types of significant multi-target-qubit phase gates useful in QIP. The proposal is quite general, which can be used to accomplish the same task for a general type of qubits such as atoms, NV centers, quantum dots, and superconducting qubits.

A high performance transparent resistive switching memory made from ZrO{sub 2}/AlON bilayer structure

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tsai, Tsung-Ling; Chang, Hsiang-Yu; Tseng, Tseung-Yuen, E-mail: tseng@cc.nctu.edu.tw

2016-04-11

In this study, the switching properties of an indium tin oxide (ITO)/zirconium oxide (ZrO{sub 2})/ITO single layer device and those of a device with an aluminum oxynitride (AlON) layer were investigated. The devices with highly transparent characteristics were fabricated. Compared with the ITO/ZrO{sub 2}/ITO single layer device, the ITO/ZrO{sub 2}/AlON/ITO bilayer device exhibited a larger ON/OFF ratio, higher endurance performance, and superior retention properties by using a simple two-step forming process. These substantial improvements in the resistive switching properties were attributed to the minimized influence of oxygen migration through the ITO top electrode (TE), which can be realized by formingmore » an asymmetrical conductive filament with the weakest part at the ZrO{sub 2}/AlON interface. Therefore, in the ITO/ZrO{sub 2}/AlON/ITO bilayer device, the regions where conductive filament formation and rupture occur can be effectively moved from the TE interface to the interior of the device.« less

Multi-target-qubit unconventional geometric phase gate in a multi-cavity system.

PubMed

Liu, Tong; Cao, Xiao-Zhi; Su, Qi-Ping; Xiong, Shao-Jie; Yang, Chui-Ping

2016-02-22

Cavity-based large scale quantum information processing (QIP) may involve multiple cavities and require performing various quantum logic operations on qubits distributed in different cavities. Geometric-phase-based quantum computing has drawn much attention recently, which offers advantages against inaccuracies and local fluctuations. In addition, multiqubit gates are particularly appealing and play important roles in QIP. We here present a simple and efficient scheme for realizing a multi-target-qubit unconventional geometric phase gate in a multi-cavity system. This multiqubit phase gate has a common control qubit but different target qubits distributed in different cavities, which can be achieved using a single-step operation. The gate operation time is independent of the number of qubits and only two levels for each qubit are needed. This multiqubit gate is generic, e.g., by performing single-qubit operations, it can be converted into two types of significant multi-target-qubit phase gates useful in QIP. The proposal is quite general, which can be used to accomplish the same task for a general type of qubits such as atoms, NV centers, quantum dots, and superconducting qubits.

A single scan skeletonization algorithm: application to medical imaging of trabecular bone

NASA Astrophysics Data System (ADS)

Arlicot, Aurore; Amouriq, Yves; Evenou, Pierre; Normand, Nicolas; Guédon, Jean-Pierre

2010-03-01

Shape description is an important step in image analysis. The skeleton is used as a simple, compact representation of a shape. A skeleton represents the line centered in the shape and must be homotopic and one point wide. Current skeletonization algorithms compute the skeleton over several image scans, using either thinning algorithms or distance transforms. The principle of thinning is to delete points as one goes along, preserving the topology of the shape. On the other hand, the maxima of the local distance transform identifies the skeleton and is an equivalent way to calculate the medial axis. However, with this method, the skeleton obtained is disconnected so it is required to connect all the points of the medial axis to produce the skeleton. In this study we introduce a translated distance transform and adapt an existing distance driven homotopic algorithm to perform skeletonization with a single scan and thus allow the processing of unbounded images. This method is applied, in our study, on micro scanner images of trabecular bones. We wish to characterize the bone micro architecture in order to quantify bone integrity.

Comparative pharmacokinetics study of orientin in rat plasma by UHPLC-MS/MS after intravenous administration of single orientin and Trollius chinensis Bunge extract.

PubMed

Zhao, Nan; Sun, Qi; Song, Yang; Wang, Lin; Zhang, Tingjian; Meng, Fanhao

2018-04-01

Orientin showed a broad array of biological activities, and it is the major bioactive compound in the Trollius chinensis Bunge. The aim of this study was to investigate the comparative pharmacokinetics of orientin after intravenous administration of single orientin and T. chinensis Bunge extract. Sample preparation involved a simple one-step deproteinization procedure with acetonitrile. Chromatographic separation was achieved on a Waters BEH C 18 column with a mobile phase consisting of acetonitrile and water containing 0.1% formic acid in an isocratic elution way. The detection was accomplished in multiple reaction monitoring mode with positive electrospray ionization. The pharmacokinetic properties of orientin were compared after intravenous administrations of pure orientin and T. chinensis Bunge extract to rats with approximately the same dosage of 10 mg/kg. The results of the study indicate that the pharmacokinetics of orientin in rat plasma show significant differences between two groups. This is useful for the clinical uses of therapeutic dosing of orientin and T. chinensis Bunge. Copyright © 2017 John Wiley & Sons, Ltd.

Simple synthetic route to manganese-containing nanowires with the spinel crystal structure

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yu, Lei; Zhang, Yan; Hudak, Bethany M.

This report describes a new route to synthesize single-crystalline manganese-containing spinel nanowires (NWs) by a two-step hydrothermal and solid-state synthesis. Interestingly, a nanowire or nanorod morphology is maintained during conversion from MnO{sub 2}/MnOOH to CuMn{sub 2}O{sub 4}/Mg{sub 2}MnO{sub 4}, despite the massive structural rearrangement this must involve. Linear sweep voltammetry (LSV) curves of the products give preliminary demonstration that CuMn{sub 2}O{sub 4} NWs are catalytically active towards the oxygen evolution reaction (OER) in alkaline solution, exhibiting five times the magnitude of current density found with pure carbon black. - Highlights: • Synthesis of single-crystalline manganese-containing spinel nanowires. • Binary oxidemore » nanowire converted to ternary oxide wire through solid state reaction. • Approach to structure conversion with shape retention could be generally applicable. • Copper and Manganese display multiple oxidation states with potential for catalysis. • CuMn{sub 2}O{sub 4} nanowires show promise as catalysts for the oxygen evolution reaction.« less

Acceleration of high-pressure-ratio single-spool turbojet engine as determined from component performance characteristics I : effect of air bleed at compressor outlet

NASA Technical Reports Server (NTRS)

Rebeske, John J , Jr; Rohlik, Harold E

1953-01-01

An analytical investigation was made to determine from component performance characteristics the effect of air bleed at the compressor outlet on the acceleration characteristics of a typical high-pressure-ratio single-spool turbojet engine. Consideration of several operating lines on the compressor performance map with two turbine-inlet temperatures showed that for a minimum acceleration time the turbine-inlet temperature should be the maximum allowable, and the operating line on the compressor map should be as close to the surge region as possible throughout the speed range. Operation along such a line would require a continuously varying bleed area. A relatively simple two-step area bleed gives only a small increase in acceleration time over a corresponding variable-area bleed. For the modes of operation considered, over 84 percent of the total acceleration time was required to accelerate through the low-speed range ; therefore, better low-speed compressor performance (higher pressure ratios and efficiencies) would give a significant reduction in acceleration time.

Single-Run Single-Mask Inductively-Coupled-Plasma Reactive-Ion-Etching Process for Fabricating Suspended High-Aspect-Ratio Microstructures

NASA Astrophysics Data System (ADS)

Yang, Yao-Joe; Kuo, Wen-Cheng; Fan, Kuang-Chao

2006-01-01

In this work, we present a single-run single-mask (SRM) process for fabricating suspended high-aspect-ratio structures on standard silicon wafers using an inductively coupled plasma-reactive ion etching (ICP-RIE) etcher. This process eliminates extra fabrication steps which are required for structure release after trench etching. Released microstructures with 120 μm thickness are obtained by this process. The corresponding maximum aspect ratio of the trench is 28. The SRM process is an extended version of the standard process proposed by BOSCH GmbH (BOSCH process). The first step of the SRM process is a standard BOSCH process for trench etching, then a polymer layer is deposited on trench sidewalls as a protective layer for the subsequent structure-releasing step. The structure is released by dry isotropic etching after the polymer layer on the trench floor is removed. All the steps can be integrated into a single-run ICP process. Also, only one mask is required. Therefore, the process complexity and fabrication cost can be effectively reduced. Discussions on each SRM step and considerations for avoiding undesired etching of the silicon structures during the release process are also presented.

A simple assay system to monitor the potential for contamination during different stages of cryopreservation.

PubMed

Morris, G J

2009-01-01

A simple assay to monitor the potential for contamination during different steps of cryopreservation is described. The assay is based on the contamination of liquid nitrogen using crystals of sucrose hemi-heptahydrate, these are stable in liquid nitrogen, nitrogen vapour and ambient air and can be monitored by a simple assay which allows contamination risks to be evaluated in a direct, rapid manner.

Errors in Postural Preparation Lead to Increased Choice Reaction Times for Step Initiation in Older Adults

PubMed Central

Nutt, John G.; Horak, Fay B.

2011-01-01

Background. This study asked whether older adults were more likely than younger adults to err in the initial direction of their anticipatory postural adjustment (APA) prior to a step (indicating a motor program error), whether initial motor program errors accounted for reaction time differences for step initiation, and whether initial motor program errors were linked to inhibitory failure. Methods. In a stepping task with choice reaction time and simple reaction time conditions, we measured forces under the feet to quantify APA onset and step latency and we used body kinematics to quantify forward movement of center of mass and length of first step. Results. Trials with APA errors were almost three times as common for older adults as for younger adults, and they were nine times more likely in choice reaction time trials than in simple reaction time trials. In trials with APA errors, step latency was delayed, correlation between APA onset and step latency was diminished, and forward motion of the center of mass prior to the step was increased. Participants with more APA errors tended to have worse Stroop interference scores, regardless of age. Conclusions. The results support the hypothesis that findings of slow choice reaction time step initiation in older adults are attributable to inclusion of trials with incorrect initial motor preparation and that these errors are caused by deficits in response inhibition. By extension, the results also suggest that mixing of trials with correct and incorrect initial motor preparation might explain apparent choice reaction time slowing with age in upper limb tasks. PMID:21498431

Prediction of work metabolism from heart rate measurements in forest work: some practical methodological issues.

PubMed

Dubé, Philippe-Antoine; Imbeau, Daniel; Dubeau, Denise; Auger, Isabelle; Leone, Mario

2015-01-01

Individual heart rate (HR) to workload relationships were determined using 93 submaximal step-tests administered to 26 healthy participants attending physical activities in a university training centre (laboratory study) and 41 experienced forest workers (field study). Predicted maximum aerobic capacity (MAC) was compared to measured MAC from a maximal treadmill test (laboratory study) to test the effect of two age-predicted maximum HR Equations (220-age and 207-0.7 × age) and two clothing insulation levels (0.4 and 0.91 clo) during the step-test. Work metabolism (WM) estimated from forest work HR was compared against concurrent work V̇O2 measurements while taking into account the HR thermal component. Results show that MAC and WM can be accurately predicted from work HR measurements and simple regression models developed in this study (1% group mean prediction bias and up to 25% expected prediction bias for a single individual). Clothing insulation had no impact on predicted MAC nor age-predicted maximum HR equations. Practitioner summary: This study sheds light on four practical methodological issues faced by practitioners regarding the use of HR methodology to assess WM in actual work environments. More specifically, the effect of wearing work clothes and the use of two different maximum HR prediction equations on the ability of a submaximal step-test to assess MAC are examined, as well as the accuracy of using an individual's step-test HR to workload relationship to predict WM from HR data collected during actual work in the presence of thermal stress.

Determining irrigation distribution uniformity and efficiency for nurseries

Treesearch

R. Thomas Fernandez

2010-01-01

A simple method for testing the distribution uniformity of overhead irrigation systems is described. The procedure is described step-by-step along with an example. Other uses of distribution uniformity testing are presented, as well as common situations that affect distribution uniformity and how to alleviate them.

Procedure for normalization of cDNA libraries

DOEpatents

Bonaldo, Maria DeFatima; Soares, Marcelo Bento

1997-01-01

This invention provides a method to normalize a cDNA library constructed in a vector capable of being converted to single-stranded circles and capable of producing complementary nucleic acid molecules to the single-stranded circles comprising: (a) converting the cDNA library in single-stranded circles; (b) generating complementary nucleic acid molecules to the single-stranded circles; (c) hybridizing the single-stranded circles converted in step (a) with complementary nucleic acid molecules of step (b) to produce partial duplexes to an appropriate Cot; (e) separating the unhybridized single-stranded circles from the hybridized single-stranded circles, thereby generating a normalized cDNA library.

Kinetics of Electrocatalytic Reactions from First-Principles: A Critical Comparison with the Ab Initio Thermodynamics Approach.

PubMed

Exner, Kai S; Over, Herbert

2017-05-16

Multielectron processes in electrochemistry require the stabilization of reaction intermediates (RI) at the electrode surface after every elementary reaction step. Accordingly, the bond strengths of these intermediates are important for assessing the catalytic performance of an electrode material. Current understanding of microscopic processes in modern electrocatalysis research is largely driven by theory, mostly based on ab initio thermodynamics considerations, where stable reaction intermediates at the electrode surface are identified, while the actual free energy barriers (or activation barriers) are ignored. This simple approach is popular in electrochemistry in that the researcher has a simple tool at hand in successfully searching for promising electrode materials. The ab initio TD approach allows for a rough but fast screening of the parameter space with low computational cost. However, ab initio thermodynamics is also frequently employed (often, even based on a single binding energy only) to comprehend on the activity and on the mechanism of an electrochemical reaction. The basic idea is that the activation barrier of an endergonic reaction step consists of a thermodynamic part and an additional kinetically determined barrier. Assuming that the activation barrier scales with thermodynamics (so-called Brønsted-Polanyi-Evans (BEP) relation) and the kinetic part of the barrier is small, ab initio thermodynamics may provide molecular insights into the electrochemical reaction kinetics. However, for many electrocatalytic reactions, these tacit assumptions are violated so that ab initio thermodynamics will lead to contradictions with both experimental data and ab initio kinetics. In this Account, we will discuss several electrochemical key reactions, including chlorine evolution (CER), oxygen evolution reaction (OER), and oxygen reduction (ORR), where ab initio kinetics data are available in order to critically compare the results with those derived from a simple ab initio thermodynamics treatment. We show that ab initio thermodynamics leads to erroneous conclusions about kinetic and mechanistic aspects for the CER over RuO 2 (110), while the kinetics of the OER over RuO 2 (110) and ORR over Pt(111) are reasonably well described. Microkinetics of an electrocatalyzed reaction is largely simplified by the quasi-equilibria of the RI preceding the rate-determining step (rds) with the reactants. Therefore, in ab initio kinetics the rate of an electrocatalyzed reaction is governed by the transition state (TS) with the highest free energy G rds # , defining also the rate-determining step (rds). Ab initio thermodynamics may be even more powerful, when using the highest free energy of an reaction intermediate G max (RI) rather than the highest free energy difference between consecutive reaction intermediates, ΔG loss , as a descriptor for the kinetics.

Science Opportunity Analyzer (SOA): Science Planning Made Simple

NASA Technical Reports Server (NTRS)

Streiffert, Barbara A.; Polanskey, Carol A.

2004-01-01

.For the first time at JPL, the Cassini mission to Saturn is using distributed science operations for developing their experiments. Remote scientists needed the ability to: a) Identify observation opportunities; b) Create accurate, detailed designs for their observations; c) Verify that their designs meet their objectives; d) Check their observations against project flight rules and constraints; e) Communicate their observations to other scientists. Many existing tools provide one or more of these functions, but Science Opportunity Analyzer (SOA) has been built to unify these tasks into a single application. Accurate: Utilizes JPL Navigation and Ancillary Information Facility (NAIF) SPICE* software tool kit - Provides high fidelity modeling. - Facilitates rapid adaptation to other flight projects. Portable: Available in Unix, Windows and Linux. Adaptable: Designed to be a multi-mission tool so it can be readily adapted to other flight projects. Implemented in Java, Java 3D and other innovative technologies. Conclusion: SOA is easy to use. It only requires 6 simple steps. SOA's ability to show the same accurate information in multiple ways (multiple visualization formats, data plots, listings and file output) is essential to meet the needs of a diverse, distributed science operations environment.

A rapid molecular diagnosis of cutaneous leishmaniasis by colorimetric malachite green-loop-mediated isothermal amplification (LAMP) combined with an FTA card as a direct sampling tool.

PubMed

Nzelu, Chukwunonso O; Cáceres, Abraham G; Guerrero-Quincho, Silvia; Tineo-Villafuerte, Edwin; Rodriquez-Delfin, Luis; Mimori, Tatsuyuki; Uezato, Hiroshi; Katakura, Ken; Gomez, Eduardo A; Guevara, Angel G; Hashiguchi, Yoshihisa; Kato, Hirotomo

2016-01-01

Leishmaniasis remains one of the world's most neglected diseases, and early detection of the infectious agent, especially in developing countries, will require a simple and rapid test. In this study, we established a quick, one-step, single-tube, highly sensitive loop-mediated isothermal amplification (LAMP) assay for rapid detection of Leishmania DNA from tissue materials spotted on an FTA card. An FTA-LAMP with pre-added malachite green was performed at 64°C for 60min using a heating block and/or water bath and DNA amplification was detected immediately after incubation. The LAMP assay had high detection sensitivity down to a level of 0.01 parasites per μl. The field- and clinic-applicability of the colorimetric FTA-LAMP assay was demonstrated with 122 clinical samples collected from patients suspected of having cutaneous leishmaniasis in Peru, from which 71 positives were detected. The LAMP assay in combination with an FTA card described here is rapid and sensitive, as well as simple to perform, and has great potential usefulness for diagnosis and surveillance of leishmaniasis in endemic areas. Copyright © 2015 Elsevier B.V. All rights reserved.

Feasibility of FT–Raman spectroscopy for rapid screening for DON toxin in ground wheat and barley.

PubMed

Liu, Y; Delwiche, S R; Dong, Y

2009-10-01

Rapid detection of deoxynivalenol (DON) in cereal-based food and feed has long been the goal of regulators and manufacturers. As non-destructive approaches, infrared (IR) and near-infrared (NIR) spectroscopic techniques have been used for the prediction and classification of contaminated single-kernel and ground grain without any DON extraction steps. These methods, however, are hindered by the intense and broad spectral bands attributed to naturally occurring moisture. Raman spectroscopy could be an alternative to IR and NIR due to its insensitivity to water and fewer overlapped bands. This study explored the feasibility of the Raman technique for rapid and non-destructive screening of DON-contaminated wheat and barley meal. The advantages of this technique include the use of a 1064-nm NIR excitation laser that reduces interference from fluorescence of biological compounds in wheat and barley, the use of a simple intensity-intensity algorithm at two unique frequencies, plus the technique's ease of sample preparation. The results indicate that the simple algorithm, as well as principal component analysis applied to the Raman spectra, can be used to classify low from high DON grain.

A simple method for finding the scattering coefficients of quantum graphs

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cottrell, Seth S.

2015-09-15

Quantum walks are roughly analogous to classical random walks, and similar to classical walks they have been used to find new (quantum) algorithms. When studying the behavior of large graphs or combinations of graphs, it is useful to find the response of a subgraph to signals of different frequencies. In doing so, we can replace an entire subgraph with a single vertex with variable scattering coefficients. In this paper, a simple technique for quickly finding the scattering coefficients of any discrete-time quantum graph will be presented. These scattering coefficients can be expressed entirely in terms of the characteristic polynomial ofmore » the graph’s time step operator. This is a marked improvement over previous techniques which have traditionally required finding eigenstates for a given eigenvalue, which is far more computationally costly. With the scattering coefficients we can easily derive the “impulse response” which is the key to predicting the response of a graph to any signal. This gives us a powerful set of tools for rapidly understanding the behavior of graphs or for reducing a large graph into its constituent subgraphs regardless of how they are connected.« less

Oscillatory magnetic tweezers based on ferromagnetic beads and simple coaxial coils

NASA Astrophysics Data System (ADS)

Trepat, Xavier; Grabulosa, Mireia; Buscemi, Lara; Rico, Fèlix; Fabry, Ben; Fredberg, Jeffrey J.; Farré, Ramon

2003-09-01

We report the design and validation of simple magnetic tweezers for oscillating ferromagnetic beads in the piconewton and nanometer scales. The system is based on a single pair of coaxial coils operating in two sequential modes: permanent magnetization of the beads through a large and brief pulse of magnetic field and generation of magnetic gradients to produce uniaxial oscillatory forces. By using this two step method, the magnetic moment of the beads remains constant during measurements. Therefore, the applied force can be computed and varies linearly with the driving signal. No feedback control is required to produce well defined force oscillations over a wide bandwidth. The design of the coils was optimized to obtain high magnetic fields (280 mT) and gradients (2 T/m) with high homogeneity (5% variation) within the sample. The magnetic tweezers were implemented in an inverted optical microscope with a videomicroscopy-based multiparticle tracking system. The apparatus was validated with 4.5 μm magnetite beads obtaining forces up to ˜2 pN and subnanometer resolution. The applicability of the device includes microrheology of biopolymer and cell cytoplasm, molecular mechanics, and mechanotransduction in living cells.

Inkjet printing of single-crystal films.

PubMed

Minemawari, Hiromi; Yamada, Toshikazu; Matsui, Hiroyuki; Tsutsumi, Jun'ya; Haas, Simon; Chiba, Ryosuke; Kumai, Reiji; Hasegawa, Tatsuo

2011-07-13

The use of single crystals has been fundamental to the development of semiconductor microelectronics and solid-state science. Whether based on inorganic or organic materials, the devices that show the highest performance rely on single-crystal interfaces, with their nearly perfect translational symmetry and exceptionally high chemical purity. Attention has recently been focused on developing simple ways of producing electronic devices by means of printing technologies. 'Printed electronics' is being explored for the manufacture of large-area and flexible electronic devices by the patterned application of functional inks containing soluble or dispersed semiconducting materials. However, because of the strong self-organizing tendency of the deposited materials, the production of semiconducting thin films of high crystallinity (indispensable for realizing high carrier mobility) may be incompatible with conventional printing processes. Here we develop a method that combines the technique of antisolvent crystallization with inkjet printing to produce organic semiconducting thin films of high crystallinity. Specifically, we show that mixing fine droplets of an antisolvent and a solution of an active semiconducting component within a confined area on an amorphous substrate can trigger the controlled formation of exceptionally uniform single-crystal or polycrystalline thin films that grow at the liquid-air interfaces. Using this approach, we have printed single crystals of the organic semiconductor 2,7-dioctyl[1]benzothieno[3,2-b][1]benzothiophene (C(8)-BTBT) (ref. 15), yielding thin-film transistors with average carrier mobilities as high as 16.4 cm(2) V(-1) s(-1). This printing technique constitutes a major step towards the use of high-performance single-crystal semiconductor devices for large-area and flexible electronics applications.

Single Molecule Stepping and Structural Dynamics of Myosin X

PubMed Central

Sun, Yujie; Sato, Osamu; Ruhnow, Felix; Arsenault, Mark E.; Ikebe, Mitsuo; Goldman, Yale E.

2010-01-01

Myosin X is an unconventional myosin with puzzling motility properties. We studied the motility of dimerized myosin X using single molecule fluorescence techniques – polTIRF, FIONA, and Parallax to measure rotation angles and 3-dimensional position of the molecule during its walk. It was found that Myosin X steps processively in a hand-over-hand manner following a left-handed helical path along both single actin filaments and bundles. Its step size and velocity are smaller on actin bundles than individual filaments, suggesting myosin X often steps onto neighboring filaments in a bundle. The data suggest that a previously postulated single α-helical domain mechanically extends the 3-IQ motif lever arm and either the neck-tail hinge or the tail is flexible. These structural features, in conjunction with the membrane and microtubule binding domains, enable myosin X to perform multiple functions on varied actin structures in cells. PMID:20364131

Using Algorithms in Solving Synapse Transmission Problems.

ERIC Educational Resources Information Center

Stencel, John E.

1992-01-01

Explains how a simple three-step algorithm can aid college students in solving synapse transmission problems. Reports that all of the students did not completely understand the algorithm. However, many learn a simple working model of synaptic transmission and understand why an impulse will pass across a synapse quantitatively. Students also see…

Modeling ultrasound propagation through material of increasing geometrical complexity.

PubMed

Odabaee, Maryam; Odabaee, Mostafa; Pelekanos, Matthew; Leinenga, Gerhard; Götz, Jürgen

2018-06-01

Ultrasound is increasingly being recognized as a neuromodulatory and therapeutic tool, inducing a broad range of bio-effects in the tissue of experimental animals and humans. To achieve these effects in a predictable manner in the human brain, the thick cancellous skull presents a problem, causing attenuation. In order to overcome this challenge, as a first step, the acoustic properties of a set of simple bone-modeling resin samples that displayed an increasing geometrical complexity (increasing step sizes) were analyzed. Using two Non-Destructive Testing (NDT) transducers, we found that Wiener deconvolution predicted the Ultrasound Acoustic Response (UAR) and attenuation caused by the samples. However, whereas the UAR of samples with step sizes larger than the wavelength could be accurately estimated, the prediction was not accurate when the sample had a smaller step size. Furthermore, a Finite Element Analysis (FEA) performed in ANSYS determined that the scattering and refraction of sound waves was significantly higher in complex samples with smaller step sizes compared to simple samples with a larger step size. Together, this reveals an interaction of frequency and geometrical complexity in predicting the UAR and attenuation. These findings could in future be applied to poro-visco-elastic materials that better model the human skull. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Selective disulfide reduction for labeling and enhancement of Fab antibody fragments

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kirley, Terence L., E-mail: terry.kirley@uc.edu; Greis, Kenneth D.; Norman, Andrew B.

Many methods have been developed for chemical labeling and enhancement of the properties of antibodies and their common fragments, including the Fab and F(ab’){sub 2} fragments. Somewhat selective reduction of some antibody disulfide bonds has been previously achieved, yielding antibodies and antibody fragments that can be labeled at defined sites, enhancing their utility and properties. Selective reduction of the two hinge disulfide bonds present in F(ab’){sub 2} fragments using mild reduction has been useful. However, such reduction is often not quantitative and results in the reduction of multiple disulfide bonds, and therefore subsequent multiple labeling or conjugation sites are neithermore » homogenous nor stoichiometric. Here, a simple and efficient selective reduction of the single disulfide bond linking the partial heavy chain and the intact light chain which compose the Fab fragment is accomplished utilizing tris(2-carboxyethyl)phosphine (TCEP) immobilized on agarose beads. The resultant reduced cysteine residues were labeled with several cysteine-selective fluorescent reagents, as well as by cysteine-directed PEGylation. These two cysteine residues can also be re-ligated by means of a bifunctional cysteine cross-linking agent, dibromobimane, thereby both restoring a covalent linkage between the heavy and light chains at this site, far removed from the antigen binding site, and also introducing a fluorescent probe. There are many other research and clinical uses for these selectively partially reduced Fab fragments, including biotinylation, toxin and drug conjugation, and incorporation of radioisotopes, and this technique enables simple generation of very useful Fab fragment derivatives with many potential applications. - Highlights: • TCEP agarose is effective for selective reduction of a single Fab disulfide bond. • This disulfide is solvent accessible and distant from the antigen binding site. • A variety of buffers of varying pHs can be used, simplifying subsequent steps. • The methods used are simple, easily verifiable, reproducible, and quantitative. • The selectively reduced Fab has many experimental and clinical applications.« less

Effects of Simple Leaching of Crushed and Powdered Materials on High-precision Pb Isotope Analyses

NASA Astrophysics Data System (ADS)

Todd, E.; Stracke, A.

2013-12-01

We present new results of simple leaching experiments on the Pb isotope composition of USGS standard reference material powders and on ocean island basalt whole rock splits and powders. Rock samples were leached with 6N HCl in two steps, first hot and then in an ultrasonic bath, and washed with ultrapure H2O before conventional sample digestion and chromatographic purification of Pb. Pb isotope analyses were determined with Tl-doped MC-ICP-MS. Intra- and inter-session analytical reproducibility of repeated analyses of both synthetic Pb solutions and Pb from single digests of chemically processed natural samples were generally < 100 ppm (2 S.D.). The comparison of leached and unleached samples shows that leaching reliably removes variable amounts of different contaminants for different starting materials. For repeated digests of a single sample, the leached samples reproduce better than the unleached ones, showing that leaching effectively removes heterogeneously distributed extraneous Pb. However, the reproducibility of repeated digests of variably contaminated natural samples is up to an order of magnitude worse than the analytical reproducibility of ca. 100 ppm. More complex leaching methods (e.g., Nobre Silva et al., 2009) yield Pb isotope ratios within error of and with similar reproducibility to our method, showing that the simple leaching method is reliable. The remaining Pb isotope heterogeneity of natural samples, which typically exceeds 100 ppm, is thus attributed to inherent isotopic sample heterogeneity. Tl-doped MC-ICP-MS Pb ratio determination is therefore a sufficiently precise method for Pb isotope analyses in natural rocks. More precise Pb double- or triple-spike methods (e.g., Galer, 1999; Thirlwall, 2000), may exploit their full potential only in cases where natural isotopic sample heterogeneity is demonstrably negligible. References: Galer, S., 1999, Chem. Geol. 157, 255-274. Nobre Silva, et al. 2009, Geochemistry Geophysics Geosystems 10, Q08012. Thirlwall, M.F., 2000, Chem. Geol. 163, 299-322.

Improved Aerogel Vacuum Thermal Insulation

NASA Technical Reports Server (NTRS)

Ruemmele, Warren P.; Bue, Grant C.

2009-01-01

An improved design concept for aerogel vacuum thermal-insulation panels calls for multiple layers of aerogel sandwiched between layers of aluminized Mylar (or equivalent) poly(ethylene terephthalate), as depicted in the figure. This concept is applicable to both the rigid (brick) form and the flexible (blanket) form of aerogel vacuum thermal-insulation panels. Heretofore, the fabrication of a typical aerogel vacuum insulating panel has involved encapsulation of a single layer of aerogel in poly(ethylene terephthalate) and pumping of gases out of the aerogel-filled volume. A multilayer panel according to the improved design concept is fabricated in basically the same way: Multiple alternating layers of aerogel and aluminized poly(ethylene terephthalate) are assembled, then encapsulated in an outer layer of poly(ethylene terephthalate), and then the volume containing the multilayer structure is evacuated as in the single-layer case. The multilayer concept makes it possible to reduce effective thermal conductivity of a panel below that of a comparable single-layer panel, without adding weight or incurring other performance penalties. Implementation of the multilayer concept is simple and relatively inexpensive, involving only a few additional fabrication steps to assemble the multiple layers prior to evacuation. For a panel of the blanket type, the multilayer concept, affords the additional advantage of reduced stiffness.

Exploiting the superior protein resistance of polymer brushes to control single cell adhesion and polarisation at the micron scale

PubMed Central

Gautrot, Julien E.; Trappmann, Britta; Oceguera-Yanez, Fabian; Connelly, John; He, Ximin; Watt, Fiona M.; Huck, Wilhelm T.S.

2010-01-01

The control of the cell microenvironment on model patterned substrates allows the systematic study of cell biology in well defined conditions, potentially using automated systems. The extreme protein resistance of poly(oligo(ethylene glycol methacrylate)) (POEGMA) brushes is exploited to achieve high fidelity patterning of single cells. These coatings can be patterned by soft lithography on large areas (a microscope slide) and scale (substrates were typically prepared in batches of 200). The present protocol relies on the adsorption of extra-cellular matrix (ECM) proteins on unprotected areas using simple incubation and washing steps. The stability of POEGMA brushes, as examined via ellipsometry and SPR, is found to be excellent, both during storage and cell culture. The impact of substrate treatment, brush thickness and incubation protocol on ECM deposition, both for ultra-thin gold and glass substrates, is investigated via fluorescence microscopy and AFM. Optimised conditions result in high quality ECM patterns at the micron scale, even on glass substrates, that are suitable for controlling cell spreading and polarisation. These patterns are compatible with state-of-the-art technologies (fluorescence microscopy, FRET) used for live cell imaging. This technology, combined with single cell analysis methods, provides a platform for exploring the mechanisms that regulate cell behaviour. PMID:20347135

Integration of strained and relaxed silicon thin films on silicon wafers via engineered oxide heterostructures: Experiment and theory

NASA Astrophysics Data System (ADS)

Seifarth, O.; Dietrich, B.; Zaumseil, P.; Giussani, A.; Storck, P.; Schroeder, T.

2010-10-01

Strained and relaxed single crystalline Si on insulator systems is an important materials science approach for future Si-based nanoelectronics. Layer transfer techniques are the dominating global integration approach over the whole wafer system but are difficult to scale down for local integration purposes limited to the area of the future device. In this respect, the heteroepitaxy approach by two simple subsequent epitaxial deposition steps of the oxide and the Si thin film is a promising way. We introduce tailored (Pr2O3)1-x(Y2O3)x oxide heterostructures on Si(111) as flexible heteroepitaxy concept for the integration of either strained or fully relaxed single crystalline Si thin films. Two different buffer concepts are explored by a combined experimental and theoretical study. First, the growth of fully relaxed single crystalline Si films is achieved by the growth of mixed PrYO3 insulators on Si(111) whose lattice constant is matched to Si. Second, isomorphic oxide-on-oxide epitaxy is exploited to grow strained Si films on lattice mismatched Y2O3/Pr2O3/Si(111) support systems. A thickness dependent multilayer model, based on Matthew's approach for strain relaxation by misfit dislocations, is presented to describe the experimental data.

Single-step solvothermal synthesis of mesoporous Ag-TiO2-reduced graphene oxide ternary composites with enhanced photocatalytic activity

NASA Astrophysics Data System (ADS)

Arif Sher Shah, Md. Selim; Zhang, Kan; Park, A. Reum; Kim, Kwang Su; Park, Nam-Gyu; Park, Jong Hyeok; Yoo, Pil J.

2013-05-01

With growing interest in the photocatalytic performance of TiO2-graphene composite systems, the ternary phase of TiO2, graphene, and Ag is expected to exhibit improved photocatalytic characteristics because of the improved recombination rate of photogenerated charge carriers and potential contribution of the generation of localized surface plasmon resonance at Ag sites on a surface of the TiO2-graphene binary matrix. In this work, Ag-TiO2-reduced graphene oxide ternary nanocomposites were successfully synthesized by a simple solvothermal process. In a single-step synthetic procedure, the reduction of AgNO3 and graphene oxide and the hydrolysis of titanium tetraisopropoxide were spontaneously performed in a mixed solvent system of ethylene glycol, N,N-dimethylformamide and a stoichiometric amount of water without resorting to the use of typical reducing agents. The nanocomposites were characterized by X-ray diffraction, X-ray photoelectron spectroscopy, along with different microscopic and spectroscopic techniques, enabling us to confirm the successful reduction of AgNO3 and graphite oxide to metallic Ag and reduced graphene oxide, respectively. Due to the highly facilitated electron transport of well distributed Ag nanoparticles, the synthesized ternary nanocomposite showed enhanced photocatalytic activity for degradation of rhodamine B dye under visible light irradiation.With growing interest in the photocatalytic performance of TiO2-graphene composite systems, the ternary phase of TiO2, graphene, and Ag is expected to exhibit improved photocatalytic characteristics because of the improved recombination rate of photogenerated charge carriers and potential contribution of the generation of localized surface plasmon resonance at Ag sites on a surface of the TiO2-graphene binary matrix. In this work, Ag-TiO2-reduced graphene oxide ternary nanocomposites were successfully synthesized by a simple solvothermal process. In a single-step synthetic procedure, the reduction of AgNO3 and graphene oxide and the hydrolysis of titanium tetraisopropoxide were spontaneously performed in a mixed solvent system of ethylene glycol, N,N-dimethylformamide and a stoichiometric amount of water without resorting to the use of typical reducing agents. The nanocomposites were characterized by X-ray diffraction, X-ray photoelectron spectroscopy, along with different microscopic and spectroscopic techniques, enabling us to confirm the successful reduction of AgNO3 and graphite oxide to metallic Ag and reduced graphene oxide, respectively. Due to the highly facilitated electron transport of well distributed Ag nanoparticles, the synthesized ternary nanocomposite showed enhanced photocatalytic activity for degradation of rhodamine B dye under visible light irradiation. Electronic supplementary information (ESI) available: XRD of GO, EDX analysis of AgTG composites, tables show the size of graphitic domains and peak area analysis, dye adsorption plot, UV-visible absorption spectra of dye at different times, and plots of ln(Ct/C0) vs. time with the corresponding fitting curves of different samples. See DOI: 10.1039/c3nr00579h

Comparison of the DNA extraction methods for polymerase chain reaction amplification from formalin-fixed and paraffin-embedded tissues.

PubMed

Sato, Y; Sugie, R; Tsuchiya, B; Kameya, T; Natori, M; Mukai, K

2001-12-01

To obtain an adequate quality and quantity of DNA from formalin-fixed and paraffin-embedded tissue, six different DNA extraction methods were compared. Four methods used deparaffinization by xylene followed by proteinase K digestion and phenol-chloroform extraction. The temperature of the different steps was changed to obtain higher yields and improved quality of extracted DNA. The remaining two methods used microwave heating for deparaffinization. The best DNA extraction method consisted of deparaffinization by microwave irradiation, protein digestion with proteinase K at 48 degrees C overnight, and no further purification steps. By this method, the highest DNA yield was obtained and the amplification of a 989-base pair beta-globin gene fragment was achieved. Furthermore, DNA extracted by means of this procedure from five gastric carcinomas was successfully used for single strand conformation polymorphism and direct sequencing assays of the beta-catenin gene. Because the microwave-based DNA extraction method presented here is simple, has a lower contamination risk, and results in a higher yield of DNA compared with the ordinary organic chemical reagent-based extraction method, it is considered applicable to various clinical and basic fields.

An approximation method for improving dynamic network model fitting.

PubMed

Carnegie, Nicole Bohme; Krivitsky, Pavel N; Hunter, David R; Goodreau, Steven M

There has been a great deal of interest recently in the modeling and simulation of dynamic networks, i.e., networks that change over time. One promising model is the separable temporal exponential-family random graph model (ERGM) of Krivitsky and Handcock, which treats the formation and dissolution of ties in parallel at each time step as independent ERGMs. However, the computational cost of fitting these models can be substantial, particularly for large, sparse networks. Fitting cross-sectional models for observations of a network at a single point in time, while still a non-negligible computational burden, is much easier. This paper examines model fitting when the available data consist of independent measures of cross-sectional network structure and the duration of relationships under the assumption of stationarity. We introduce a simple approximation to the dynamic parameters for sparse networks with relationships of moderate or long duration and show that the approximation method works best in precisely those cases where parameter estimation is most likely to fail-networks with very little change at each time step. We consider a variety of cases: Bernoulli formation and dissolution of ties, independent-tie formation and Bernoulli dissolution, independent-tie formation and dissolution, and dependent-tie formation models.

Impaired protein conformational landscapes as revealed in anomalous Arrhenius prefactors.

PubMed

Nagel, Zachary D; Dong, Ming; Bahnson, Brian J; Klinman, Judith P

2011-06-28

A growing body of data supports a role for protein motion in enzyme catalysis. In particular, the ability of enzymes to sample catalytically relevant conformational substates has been invoked to model kinetic and spectroscopic data. However, direct experimental links between rapidly interconverting conformations and the chemical steps of catalysis remain rare. We report here on the kinetic analysis and characterization of the hydride transfer step catalyzed by a series of mutant thermophilic alcohol dehydrogenases (ht-ADH), presenting evidence for Arrhenius prefactor values that become enormously elevated above an expected value of approximately 10(13) s(-1) when the enzyme operates below its optimal temperature range. Restoration of normal Arrhenius behavior in the ht-ADH reaction occurs at elevated temperatures. A simple model, in which reduced temperature alters the ability of the ht-ADH variants to sample the catalytically relevant region of conformational space, can reproduce the available data. These findings indicate an impaired landscape that has been generated by the combined condition of reduced temperature and mutation at a single, active-site hydrophobic side chain. The broader implication is that optimal enzyme function requires the maintenance of a relatively smooth landscape that minimizes low energy traps.

Impaired protein conformational landscapes as revealed in anomalous Arrhenius prefactors

PubMed Central

Nagel, Zachary D.; Dong, Ming; Bahnson, Brian J.; Klinman, Judith P.

2011-01-01

A growing body of data supports a role for protein motion in enzyme catalysis. In particular, the ability of enzymes to sample catalytically relevant conformational substates has been invoked to model kinetic and spectroscopic data. However, direct experimental links between rapidly interconverting conformations and the chemical steps of catalysis remain rare. We report here on the kinetic analysis and characterization of the hydride transfer step catalyzed by a series of mutant thermophilic alcohol dehydrogenases (ht-ADH), presenting evidence for Arrhenius prefactor values that become enormously elevated above an expected value of approximately 1013 s-1 when the enzyme operates below its optimal temperature range. Restoration of normal Arrhenius behavior in the ht-ADH reaction occurs at elevated temperatures. A simple model, in which reduced temperature alters the ability of the ht-ADH variants to sample the catalytically relevant region of conformational space, can reproduce the available data. These findings indicate an impaired landscape that has been generated by the combined condition of reduced temperature and mutation at a single, active-site hydrophobic side chain. The broader implication is that optimal enzyme function requires the maintenance of a relatively smooth landscape that minimizes low energy traps. PMID:21670258

Computational Phenotyping in Psychiatry: A Worked Example

PubMed Central

2016-01-01

Abstract Computational psychiatry is a rapidly emerging field that uses model-based quantities to infer the behavioral and neuronal abnormalities that underlie psychopathology. If successful, this approach promises key insights into (pathological) brain function as well as a more mechanistic and quantitative approach to psychiatric nosology—structuring therapeutic interventions and predicting response and relapse. The basic procedure in computational psychiatry is to build a computational model that formalizes a behavioral or neuronal process. Measured behavioral (or neuronal) responses are then used to infer the model parameters of a single subject or a group of subjects. Here, we provide an illustrative overview over this process, starting from the modeling of choice behavior in a specific task, simulating data, and then inverting that model to estimate group effects. Finally, we illustrate cross-validation to assess whether between-subject variables (e.g., diagnosis) can be recovered successfully. Our worked example uses a simple two-step maze task and a model of choice behavior based on (active) inference and Markov decision processes. The procedural steps and routines we illustrate are not restricted to a specific field of research or particular computational model but can, in principle, be applied in many domains of computational psychiatry. PMID:27517087

Core-shell TiO2@ZnO nanorods for efficient ultraviolet photodetection.

PubMed

Panigrahi, Shrabani; Basak, Durga

2011-05-01

Core-shell TiO(2)@ZnO nanorods (NRs) have been fabricated by a simple two step method: growth of ZnO NRs' array by an aqueous chemical technique and then coating of the NRs with a solution of titanium isopropoxide [Ti(OC(3)H(7))(4)] followed by a heating step to form the shell. The core-shell nanocomposites are composed of single-crystalline ZnO NRs, coated with a thin TiO(2) shell layer obtained by varying the number of coatings (one, three and five times). The ultraviolet (UV) emission intensity of the nanocomposite is largely quenched due to an efficient electron-hole separation reducing the band-to-band recombinations. The UV photoconductivity of the core-shell structure with three times TiO(2) coating has been largely enhanced due to photoelectron transfer between the core and the shell. The UV photosensitivity of the nanocomposite becomes four times larger while the photocurrent decay during steady UV illumination has been decreased almost by 7 times compared to the as-grown ZnO NRs indicating high efficiency of these core-shell structures as UV sensors. © The Royal Society of Chemistry 2011

Clean-up of aqueous acetone vegetable extracts by solid-matrix partition for pyrethroid residue determination by gas chromatography-electron-capture detection.

PubMed

Di Muccio, A; Barbini, D A; Generali, T; Pelosi, P; Ausili, A; Vergori, F; Camoni, I

1997-03-21

Disposable, ready-to-use cartridges filled with macroporous diatomaceous material are used to carry out a partition clean-up that, in a single step, is capable of transferring pesticide residues from aqueous acetone extracts into light petroleum-dichloromethane (75:25, v/v). This procedure takes the place of some functions (such as separatory-funnel partition, drying over anhydrous sodium sulphate and partial adsorption clean-up) usually performed by separate steps in classical schemes. Fourteen pyrethroid pesticides, including tefluthrin, tetramethrin, cyphenothrin, cyfluthrin, flucythrinate, tau-fluvalinate, deltamethrin, bioallethrin, fenpropathrin, lambda-cyhalothrin, permethrin, alpha-cypermethrin, esfenvalerate and tralomethrin were determined using the described procedure with satisfactory recoveries for most of them, at spiking levels ranging from 0.08 to 0.82 mg/kg for the different compounds. Crops subjected to the described procedure included strawberry, apple, and orange gave extracts containing a mass of co-extractives that was between 5 and 30 mg. Compared with classical schemes, the described procedure is simple, less labour intensive, allows parallel handling of several extracts and does not require the preparation and maintenance of equipment. Troublesome emulsions such as those frequently observed in separation funnel partitioning do not occur.

M13 bacteriophage purification using poly(ionic liquids) as alternative separation matrices.

PubMed

Jacinto, Maria João; Patinha, David J S; Marrucho, Isabel M; Gonçalves, João; Willson, Richard C; Azevedo, Ana M; Aires-Barros, M Raquel

2018-01-12

M13 is a filamentous, non-lytic bacteriophage that infects Escherichia coli via the F pilus. Currently, phage M13 is widely used in phage display technology and bio-nanotechnology, and is considered a possible antibacterial therapeutic agent, among other applications. Conventional phage purification involves 5-7 operational steps, with high operational costs and significant product loss (approximately 60%). In this work, we propose a scalable purification process for M13 bacteriophage using a novel stationary phase based on a polymeric ionic liquid (PIL) with a positively charged backbone structure. Poly (1-vinyl-3-ethyl imidazolium bis(trifluoromethylsulfonyl) imide) - poly(VEIM-TFSI) predominantly acted as an anion exchanger under binding-elution mode. This revealed to be a rapid and simple method for the recovery of phage M13 with an overall separation yield of over 70% after a single downstream step. To the best of our knowledge, PILs have never been used as separation matrices for biological products and the results obtained, together with the large number of cations and anions available to prepare PILs, illustrate well the large potential of the proposed methodology. Copyright © 2017 Elsevier B.V. All rights reserved.

Computational Phenotyping in Psychiatry: A Worked Example.

PubMed

Schwartenbeck, Philipp; Friston, Karl

2016-01-01

Computational psychiatry is a rapidly emerging field that uses model-based quantities to infer the behavioral and neuronal abnormalities that underlie psychopathology. If successful, this approach promises key insights into (pathological) brain function as well as a more mechanistic and quantitative approach to psychiatric nosology-structuring therapeutic interventions and predicting response and relapse. The basic procedure in computational psychiatry is to build a computational model that formalizes a behavioral or neuronal process. Measured behavioral (or neuronal) responses are then used to infer the model parameters of a single subject or a group of subjects. Here, we provide an illustrative overview over this process, starting from the modeling of choice behavior in a specific task, simulating data, and then inverting that model to estimate group effects. Finally, we illustrate cross-validation to assess whether between-subject variables (e.g., diagnosis) can be recovered successfully. Our worked example uses a simple two-step maze task and a model of choice behavior based on (active) inference and Markov decision processes. The procedural steps and routines we illustrate are not restricted to a specific field of research or particular computational model but can, in principle, be applied in many domains of computational psychiatry.

Purification of circular DNA using benzoylated naphthoylated DEAE-cellulose.

PubMed

Gamper, H; Lehman, N; Piette, J; Hearst, J E

1985-04-01

Un-nicked circular DNA can be separated from protein, RNA, and other DNA in a simple three-step protocol consisting of exonuclease III digestion, extraction with benzoylated naphthoylated DEAE-cellulose (BND cellulose) in 1 M NaCl, and alcohol precipitation of the remaining supercoiled DNA. Exonuclease III treatment introduces single-stranded regions into contaminating linear and nicked circular DNA. This DNA, together with most RNA and protein, is adsorbed onto BND cellulose leaving form I DNA in solution. The protocol can be used to purify analytical as well as preparative amounts of supercoiled DNA. This procedure is a substitute for cesium chloride-ethidium bromide gradient ultracentrifugation and gives a comparable yield of pure form I DNA. Other classes of DNA can be isolated by changing the pretreatment step. Selective digestion of linear DNA with lambda exonuclease permits the isolation of both nicked circular and supercoiled DNA while brief heat-induced or alkali-induced denaturation leads to the recovery of rapidly reannealing DNA. In large-scale purifications, the basic protocol is usually preceded by one or more BND cellulose extractions in 1 M NaCl to remove contaminants absorbing UV or inhibiting exonuclease III.

Balancing Chemical Equations: The Role of Developmental Level and Mental Capacity.

ERIC Educational Resources Information Center

Niaz, Mansoor; Lawson, Anton E.

1985-01-01

Tested two hypotheses: (1) formal reasoning is required to balance simple one-step equations; and (2) formal reasoning plus sufficient mental capacity are required to balance many-step equations. Independent variables included intellectual development, mental capacity, and degree of field dependence/independence. With 25 subjects, significance was…

Kitchen Microbiology: It's Easier Than You Think!

ERIC Educational Resources Information Center

Wilcoxson, Catherine; Shand, Stacey M.; Shand, Richard F.

1999-01-01

Offers a simple experiment that tests the effectiveness of cleaners that inhibit bacterial growth. Outlines a step-by-step method for preparing culture media that can be used for isolation and propagation of microorganisms. Concludes that kitchen medium is as efficient as the professionally-made medium in supporting colony growth. (CCM)

Voicing Support for the Arts.

ERIC Educational Resources Information Center

Arts Midwest, Minneapolis, MN.

This report proposes 40 specific steps that can be taken to strengthen arts advocacy efforts in the region of the United States. Some steps can be taken immediately; others may take time to realize. There are five primary areas of recommended activities in this strategy, with simple "how to get started" instructions: (1)…

Wilderness education: The ultimate commitment to quality wilderness stewardship

Treesearch

Gregory F. Hansen; Tom Carlson

2007-01-01

The effective planning, implementation, and monitoring of a wilderness education program will ultimately produce measurable results that can be instrumental in achieving wilderness management goals and objectives. This paper will provide a simple step-by-step overview of how to develop and maintain a successful wilderness education program through planning,...

Navigating the Grad School Application Process: A Training Schedule

ERIC Educational Resources Information Center

Swindlehurst, Garrett R.; Bullard, Lisa G.

2014-01-01

Through a simple step-by-step guide for navigating the graduate school application process, a graduate student who's been through the ringer and a faculty advisor who knows the ropes offer advice to walk prospective grad students through the process of successfully entering graduate school. A repeat printing.

Getting a Head Start on Prewriting Skills.

ERIC Educational Resources Information Center

Armfield, Marge

Although handwriting should not be formally taught in the preschool years, teachers can help children develop visual motor integration and other related skills required for writing. The activities suggested in this manual, while neither new nor different, are presented in a simple step-by-step form designed to facilitate success. Exercises focus…

The Practical Search

ERIC Educational Resources Information Center

Hamilton, John D.

2007-01-01

Although the five classic evidence-based medicine steps to make the literature useful in caring for patients sound simple, most clinicians who try this approach do not find it easy. Although the first step--creating an answerable question--is manageable for busy clinicians, tracking down "the best evidence" and critically appraising it for…

Apples, Bubbles, and Crystals: Your Science ABCs.

ERIC Educational Resources Information Center

Bennett, Andrea T.; Kessler, James H.

In this book a character named Archie and his friends teach science and reinforce alphabet skills. This approach combines reading and rhyming with simple science activities that begin with a list of everyday household materials and come with colorfully illustrated step-by-step instructions. Also provided are explanations of the science principles…

Influence of dose rate on the induction of simple and complex chromosome exchanges by gamma rays.

PubMed

Loucas, Bradford D; Eberle, Richard; Bailey, Susan M; Cornforth, Michael N

2004-10-01

Single-color painting of whole chromosomes, or protocols in which only a few chromosomes are distinctively painted, will always fail to detect a proportion of complex exchanges because they frequently produce pseudosimple painting patterns that are indistinguishable from those produced by bona fide simple exchanges. When 24-color multi-fluor FISH (mFISH) was employed for the purpose of distinguishing (truly) simple from pseudosimple exchanges, it was confirmed that the acute low-LET radiation dose-response relationship for simple exchanges lacked significant upward curvature. This result has been interpreted to indicate that the formation of simple exchanges requires only one chromosome locus be damaged (e.g. broken) by radiation to initiate an exchange-not two, as classical cytogenetic theory maintains. Because a one-lesion mechanism implies single-track action, it follows that the production of simple exchanges should not be influenced by changes in dose rate. To examine this prediction, we irradiated noncycling primary human fibroblasts with graded doses of (137)Cs gamma rays at an acute dose rate of 1.10 Gy/min and compared, using mFISH, the yield of simple exchanges to that observed after exposure to the same radiation delivered at a chronic dose rate of 0.08 cGy/min. The shape of the dose response was found to be quasi-linear for both dose rates, but, counter to providing support for a one-lesion mechanism, the yield of simple aberrations was greatly reduced by protracted exposure. Although chronic doses were delivered at rates low enough to produce damage exclusively by single-track action, this did not altogether eliminate the formation of complex aberrations, an analysis of which leads to the conclusion that a single track of low-LET radiation is capable of inducing complex exchanges requiring up to four proximate breaks for their formation. For acute exposures, the ratio of simple reciprocal translocations to simple dicentrics was near unity.

Differences in Lower Extremity and Trunk Kinematics between Single Leg Squat and Step Down Tasks

PubMed Central

Lewis, Cara L.; Foch, Eric; Luko, Marc M.; Loverro, Kari L.; Khuu, Anne

2015-01-01

The single leg squat and single leg step down are two commonly used functional tasks to assess movement patterns. It is unknown how kinematics compare between these tasks. The purpose of this study was to identify kinematic differences in the lower extremity, pelvis and trunk between the single leg squat and the step down. Fourteen healthy individuals participated in this research and performed the functional tasks while kinematic data were collected for the trunk, pelvis, and lower extremities using a motion capture system. For the single leg squat task, the participant was instructed to squat as low as possible. For the step down task, the participant was instructed to stand on top of a box, slowly lower him/herself until the non-stance heel touched the ground, and return to standing. This was done from two different heights (16cm and 24cm). The kinematics were evaluated at peak knee flexion as well as at 60° of knee flexion. Pearson correlation coefficients (r) between the angles at those two time points were also calculated to better understand the relationship between each task. The tasks resulted in kinematics differences at the knee, hip, pelvis, and trunk at both time points. The single leg squat was performed with less hip adduction (p ≤ 0.003), but more hip external rotation and knee abduction (p ≤ 0.030), than the step down tasks at 60° of knee flexion. These differences were maintained at peak knee flexion except hip external rotation was only significant in the 24cm step down task (p ≤ 0.029). While there were multiple differences between the two step heights at peak knee flexion, the only difference at 60° of knee flexion was in trunk flexion (p < 0.001). Angles at the knee and hip had a moderate to excellent correlation (r = 0.51–0.98), but less consistently so at the pelvis and trunk (r = 0.21–0.96). The differences in movement patterns between the single leg squat and the step down should be considered when selecting a single leg task for evaluation or treatment. The high correlation of knee and hip angles between the three tasks indicates that similar information about knee and hip kinematics was gained from each of these tasks, while pelvis and trunk angles were less well predicted. PMID:25955321

Synthesis of ST7612AA1, a Novel Oral HDAC Inhibitor, via Radical 
Thioacetic Acid Addition.

PubMed

Battistuzzi, Gianfranco; Giannini, Giuseppe

2016-12-01

In the expanding field of anticancer drugs, HDAC inhibitors are playing an increasingly important role. To date, four/five HDAC inhibitors have been approved by FDA. All these compounds fit the widely accepted HDAC inhibitors pharmacophore model characterized by a cap group, a linker chain and a zinc binding group (ZBG), able to bind the Zn 2+ ion in a pocket of the HDAC active site. Romidepsin, a natural compound, is the only thiol derivative. We have selected a new class of synthetic HDAC inhibitors, the thio-ω(lactam-carboxamide) derivatives, with ST7612AA1 as drug candidate, pan-inhibitor active in the range of single- to two-digit nanomolar concentrations. Preliminary results of a synthetic optimization attempt towards a fast scale-up process are here proposed. In the four steps of synthesis, from unsaturated amino acid intermediate to the final product, we explored different synthetic conditions in order to have a transferable process for a scale-up synthetic laboratory. In the first step, isobutyl chloroformate was used and, after a simple work up with 1M HCl, 2 (96% yield) was obtained as a white solid, which was used directly in the next step. For thioacetic acid addition to the double bond of intermediate 2 , two different routes were possible, with addition reaction in the first (D') or last step (D). Reactions of 2 to give 5 or of 4 to give ST7612AA1 were both performed in dioxane. Reactions were fast and did not need the usually advised radical quenching with cyclohexene. The corresponding products were obtained in good yields (step D', 89%; step D, 81%) after a flash chromatography. , a thiol derivative prodrug of ST7464AA1 , is the first of a new generation of HDAC inhibitors, very potent, orally administered, and well tolerated. Here, we have identified a synthetic route, competitive, versatile and easily transferable to industrial processes.

Transportable Maps Software. Volume I.

DTIC Science & Technology

1982-07-01

being collected at the beginning or end of the routine. This allows the interaction to be followed sequentially through its steps by anyone reading the...flow is either simple sequential , simple conditional (the equivalent of ’if-then-else’), simple iteration (’DO-loop’), or the non-linear recursion...input raster images to be in the form of sequential binary files with a SEGMENTED record type. The advantage of this form is that large logical records

Magnetically-focusing biochip structures for high-speed active biosensing with improved selectivity.

PubMed

Yoo, Haneul; Lee, Dong Jun; Kim, Daesan; Park, Juhun; Chen, Xing; Hong, Seunghun

2018-06-29

We report a magnetically-focusing biochip structure enabling a single layered magnetic trap-and-release cycle for biosensors with an improved detection speed and selectivity. Here, magnetic beads functionalized with specific receptor molecules were utilized to trap target molecules in a solution and transport actively to and away from the sensor surfaces to enhance the detection speed and reduce the non-specific bindings, respectively. Using our method, we demonstrated the high speed detection of IL-13 antigens with the improved detection speed by more than an order of magnitude. Furthermore, the release step in our method was found to reduce the non-specific bindings and improve the selectivity and sensitivity of biosensors. This method is a simple but powerful strategy and should open up various applications such as ultra-fast biosensors for point-of-care services.

The Domino Way to Heterocycles

PubMed Central

Padwa, Albert; Bur, Scott K.

2007-01-01

Sequential transformations enable the facile synthesis of complex target molecules from simple building blocks in a single preparative step. Their value is amplified if they also create multiple stereogenic centers. In the ongoing search for new domino processes, emphasis is usually placed on sequential reactions which occur cleanly and without forming by-products. As a prerequisite for an ideally proceeding one-pot sequential transformation, the reactivity pattern of all participating components has to be such that each building block gets involved in a reaction only when it is supposed to do so. The development of sequences that combine transformations of fundamentally different mechanisms broadens the scope of such procedures in synthetic chemistry. This mini review contains a representative sampling from the last 15 years on the kinds of reactions that have been sequenced into cascades to produce heterocyclic molecules. PMID:17940591

Removing ring artefacts from synchrotron radiation-based hard x-ray tomography data

NASA Astrophysics Data System (ADS)

Thalmann, Peter; Bikis, Christos; Schulz, Georg; Paleo, Pierre; Mirone, Alessandro; Rack, Alexander; Siegrist, Stefan; Cörek, Emre; Huwyler, Jörg; Müller, Bert

2017-09-01

In hard X-ray microtomography, ring artefacts regularly originate from incorrectly functioning pixel elements on the detector or from particles and scratches on the scintillator. We show that due to the high sensitivity of contemporary beamline setups further causes inducing inhomogeneities in the impinging wavefronts have to be considered. We propose in this study a method to correct the thereby induced failure of simple flatfield approaches. The main steps of the pipeline are (i) registration of the reference images with the radiographs (projections), (ii) integration of the flat-field corrected projection over the acquisition angle, (iii) high-pass filtering of the integrated projection, (iv) subtraction of filtered data from the flat-field corrected projections. The performance of the protocol is tested on data sets acquired at the beamline ID19 at ESRF using single distance phase tomography.

Magnetically-focusing biochip structures for high-speed active biosensing with improved selectivity

NASA Astrophysics Data System (ADS)

Yoo, Haneul; Lee, Dong Jun; Kim, Daesan; Park, Juhun; Chen, Xing; Hong, Seunghun

2018-06-01

We report a magnetically-focusing biochip structure enabling a single layered magnetic trap-and-release cycle for biosensors with an improved detection speed and selectivity. Here, magnetic beads functionalized with specific receptor molecules were utilized to trap target molecules in a solution and transport actively to and away from the sensor surfaces to enhance the detection speed and reduce the non-specific bindings, respectively. Using our method, we demonstrated the high speed detection of IL-13 antigens with the improved detection speed by more than an order of magnitude. Furthermore, the release step in our method was found to reduce the non-specific bindings and improve the selectivity and sensitivity of biosensors. This method is a simple but powerful strategy and should open up various applications such as ultra-fast biosensors for point-of-care services.

Rapid Synthesis of 68Ga-labeled macroaggregated human serum albumin (MAA) for routine application in perfusion imaging using PET/CT.

PubMed

Mueller, D; Kulkarni, Harshad; Baum, Richard P; Odparlik, Andreas

2017-04-01

99m Tc-labeled MAA is commonly used for single photon emission computed tomography SPECT. In contrast, positron emission tomography/CT (PET/CT) delivers images with significantly higher resolution. The generator produced radionuclide 68 Ga is widely used for PET/CT imaging agents and 68 Ga-labeled MAA represents an attractive alternative to 99m Tc-labeled MAA. We report a simple and rapid NaCl based labeling procedure for the labeling of MAA with 68 Ga using a commercially available MAA labeling kit for 99m Tc. The procedure delivers 68 Ga-labeled MAA with a high specific activity and a high labeling efficiency (>99%). The synthesis does not require a final step of separation or the use of organic solvents. Copyright © 2017 Elsevier Ltd. All rights reserved.

Photochemical Approaches to Complex Chemotypes: Applications in Natural Product Synthesis

PubMed Central

2016-01-01

The use of photochemical transformations is a powerful strategy that allows for the formation of a high degree of molecular complexity from relatively simple building blocks in a single step. A central feature of all light-promoted transformations is the involvement of electronically excited states, generated upon absorption of photons. This produces transient reactive intermediates and significantly alters the reactivity of a chemical compound. The input of energy provided by light thus offers a means to produce strained and unique target compounds that cannot be assembled using thermal protocols. This review aims at highlighting photochemical transformations as a tool for rapidly accessing structurally and stereochemically diverse scaffolds. Synthetic designs based on photochemical transformations have the potential to afford complex polycyclic carbon skeletons with impressive efficiency, which are of high value in total synthesis. PMID:27120289

MemStar: a one-shot Escherichia coli-based approach for high-level bacterial membrane protein production.

PubMed

Lee, Chiara; Kang, Hae Joo; Hjelm, Anna; Qureshi, Abdul Aziz; Nji, Emmanuel; Choudhury, Hassanul; Beis, Konstantinos; de Gier, Jan-Willem; Drew, David

2014-10-16

Optimising membrane protein production yields in Escherichiacoli can be time- and resource-consuming. Here, we present a simple and effective Membrane protein Single shot amplification recipe: MemStar. This one-shot amplification recipe is based on the E. coli strain Lemo21(DE3), the PASM-5052 auto-induction medium and, contradictorily, an IPTG induction step. Using MemStar, production yields for most bacterial membrane proteins tested were improved to reach an average of 5 mg L(-1) per OD600 unit, which is significantly higher than yields obtained with other common production strategies. With MemStar, we have been able to obtain new structural information for several transporters, including the sodium/proton antiporter NapA. Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Isogenic transgenic homozygous fish induced by artificial parthenogenesis.

PubMed

Nam, Y K; Cho, Y S; Kim, D S

2000-12-01

As a model system for vertebrate transgenesis, fish have many attractive advantages, especially with respect to the characteristics of eggs, allowing us to produce isogenic, transgenic, homozygous vertebrates by combining with chromosome-set manipulation. Here, we describe the large-scale production of isogenic transgenic homozygous animals using our experimental organism, the mud loach Misgurnus mizolepis, by the simple process of artificial parthenogenesis in a single generation. These isogenic fish have retained transgenic homozygous status in a stable manner during the subsequent 5 years, and exhibited increased levels of transgene expression. Furthermore, their isogenic nature was confirmed by cloned transgenic homozygous offspring produced via another step of parthenogenic reproduction of the isogenic homozygous transgenic fish. These results demonstrate that a combination of transgenesis and artificial parthenogenesis will make the rapid utilization of genetically pure homozygous transgenic system in vertebrate transgenesis possible.

A Controlled-Phase Gate via Adiabatic Rydberg Dressing of Neutral Atoms

NASA Astrophysics Data System (ADS)

Keating, Tyler; Deutsch, Ivan; Cook, Robert; Biederman, Grant; Jau, Yuan-Yu

2014-05-01

The dipole blockade effect between Rydberg atoms is a promising tool for quantum information processing in neutral atoms. So far, most efforts to perform a quantum logic gate with this effect have used resonant laser pulses to excite the atoms, which makes the system particularly susceptible to decoherence through thermal motional effects. We explore an alternative scheme in which the atomic ground states are adiabatically ``dressed'' by turning on an off-resonant laser. We analyze the implementation of a CPHASE gate using this mechanism and find that fidelities of >99% should be possible with current technology, owing primarily to the suppression of motional errors. We also discuss how such a scheme could be generalized to perform more complicated, multi-qubit gates; in particular, a simple generalization would allow us to perform a Toffoli gate in a single step.

Surface modification of polyester synthetic leather with tetramethylsilane by atmospheric pressure plasma

NASA Astrophysics Data System (ADS)

Kan, C. W.; Kwong, C. H.; Ng, S. P.

2015-08-01

Much works have been done on synthetic materials but scarcely on synthetic leather owing to its surface structures in terms of porosity and roughness. This paper examines the use of atmospheric pressure plasma (APP) treatment for improving the surface performance of polyester synthetic leather by use of a precursor, tetramethylsilane (TMS). Plasma deposition is regarded as an effective, simple and single-step method with low pollution. Scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR) confirm the deposition of organosilanes on the sample's surface. The results showed that under a particular combination of treatment parameters, a hydrophobic surface was achieved on the APP treated sample with sessile drop static contact angle of 138°. The hydrophobic surface is stable without hydrophilic recovery 30 days after plasma treatment.

An infinite-order two-component relativistic Hamiltonian by a simple one-step transformation.

PubMed

Ilias, Miroslav; Saue, Trond

2007-02-14

The authors report the implementation of a simple one-step method for obtaining an infinite-order two-component (IOTC) relativistic Hamiltonian using matrix algebra. They apply the IOTC Hamiltonian to calculations of excitation and ionization energies as well as electric and magnetic properties of the radon atom. The results are compared to corresponding calculations using identical basis sets and based on the four-component Dirac-Coulomb Hamiltonian as well as Douglas-Kroll-Hess and zeroth-order regular approximation Hamiltonians, all implemented in the DIRAC program package, thus allowing a comprehensive comparison of relativistic Hamiltonians within the finite basis approximation.

Acceptance and introduction of disruptive technologies - simple steps to build a fully functional pulmonary valved stent.

PubMed

Huber, Christoph H; Marty, Bettina; von Segesser, Ludwig K

2007-08-01

Valved stents are new land for cardiac surgeons even though they are being used more frequently by interventional disciplines. This paper presents simple steps to build a patient-specific pulmonary valved stent and its delivery device. The design concept was tested by random participants at a med-tech meeting. The valved stent is constructed by linking an endoprosthetic graft with a valved-jugular-vein. The delivery device is made from a modified 5-ml syringe. Of 72 participants, 66 (92%) built and 60 participants implanted the device successfully into the targeted pulmonary position via a trans-infundibular access.

Facile generation of cell microarrays using vacuum degassing and coverslip sweeping.

PubMed

Wang, Min S; Luo, Zhen; Cherukuri, Sundar; Nitin, Nitin

2014-07-15

A simple method to generate cell microarrays with high-percentage well occupancy and well-defined cell confinement is presented. This method uses a synergistic combination of vacuum degassing and coverslip sweeping. The vacuum degassing step dislodges air bubbles from the microwells, which in turn enables the cells to enter the microwells, while the physical sweeping step using a glass coverslip removes the excess cells outside the microwells. This low-cost preparation method provides a simple solution to generating cell microarrays that can be performed in basic research laboratories and point-of-care settings for routine cell-based screening assays. Copyright © 2014 Elsevier Inc. All rights reserved.

A PDMS/paper/glass hybrid microfluidic biochip integrated with aptamer-functionalized graphene oxide nano-biosensors for one-step multiplexed pathogen detection.

PubMed

Zuo, Peng; Li, XiuJun; Dominguez, Delfina C; Ye, Bang-Ce

2013-10-07

Infectious pathogens often cause serious public health concerns throughout the world. There is an increasing demand for simple, rapid and sensitive approaches for multiplexed pathogen detection. In this paper we have developed a polydimethylsiloxane (PDMS)/paper/glass hybrid microfluidic system integrated with aptamer-functionalized graphene oxide (GO) nano-biosensors for simple, one-step, multiplexed pathogen detection. The paper substrate used in this hybrid microfluidic system facilitated the integration of aptamer biosensors on the microfluidic biochip, and avoided complicated surface treatment and aptamer probe immobilization in a PDMS or glass-only microfluidic system. Lactobacillus acidophilus was used as a bacterium model to develop the microfluidic platform with a detection limit of 11.0 cfu mL(-1). We have also successfully extended this method to the simultaneous detection of two infectious pathogens - Staphylococcus aureus and Salmonella enterica. This method is simple and fast. The one-step 'turn on' pathogen assay in a ready-to-use microfluidic device only takes ~10 min to complete on the biochip. Furthermore, this microfluidic device has great potential in rapid detection of a wide variety of different other bacterial and viral pathogens.

A PDMS/paper/glass hybrid microfluidic biochip integrated with aptamer-functionalized graphene oxide nano-biosensors for one-step multiplexed pathogen detection

PubMed Central

Zuo, Peng; Dominguez, Delfina C.; Ye, Bang-Ce

2014-01-01

Infectious pathogens often cause serious public health concerns throughout the world. There is an increasing demand for simple, rapid and sensitive approaches for multiplexed pathogen detection. In this paper we have developed a polydimethylsiloxane (PDMS)/paper/glass hybrid microfluidic system integrated with aptamer-functionalized graphene oxide (GO) nano-biosensors for simple, one-step, multiplexed pathogen detection. The paper substrate used in this hybrid microfluidic system facilitated the integration of aptamer biosensors on the microfluidic biochip, and avoided complicated surface treatment and aptamer probe immobilization in a PDMS or glass-only microfluidic system. Lactobacillus acidophilus was used as a bacterium model to develop the microfluidic platform with a detection limit of 11.0 cfu mL−1. We have also successfully extended this method to the simultaneous detection of two infectious pathogens - Staphylococcus aureus and Salmonella enterica. This method is simple and fast. The one-step ‘turn on’ pathogen assay in a ready-to-use microfluidic device only takes ~10 min to complete on the biochip. Furthermore, this microfluidic device has great potential in rapid detection of a wide variety of different other bacterial and viral pathogens. PMID:23929394

Quenching of unincorporated amplification signal reporters in reverse-transcription loop-mediated isothermal amplification enabling bright, single-step, closed-tube, and multiplexed detection of RNA viruses

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ball, Cameron S.; Light, Yooli K.; Koh, Chung -Yan

Reverse-transcription-loop-mediated isothermal amplification (RT-LAMP) has frequently been proposed as an enabling technology for simplified diagnostic tests for RNA viruses. However, common detection techniques used for LAMP and RT-LAMP have drawbacks, including poor discrimination capability, inability to multiplex targets, high rates of false positives, and (in some cases) the requirement of opening reaction tubes postamplification. Here, we present a simple technique that allows closed-tube, target-specific detection, based on inclusion of a dye-labeled primer that is incorporated into a target-specific amplicon if the target is present. A short, complementary quencher hybridizes to unincorporated primer upon cooling down at the end of themore » reaction, thereby quenching fluorescence of any unincorporated primer. Our technique, which we term QUASR (for quenching of unincorporated amplification signal reporters, read “quasar”), does not significantly reduce the amplification efficiency or sensitivity of RT-LAMP. Equipped with a simple LED excitation source and a colored plastic gel filter, the naked eye or a camera can easily discriminate between positive and negative QUASR reactions, which produce a difference in signal of approximately 10:1 without background subtraction. We demonstrate that QUASR detection is compatible with complex sample matrices such as human blood, using a novel LAMP primer set for bacteriophage MS2 (a model RNA virus particle). As a result, we demonstrate single-tube duplex detection of West Nile virus (WNV) and chikungunya virus (CHIKV) RNA.« less

Quenching of unincorporated amplification signal reporters in reverse-transcription loop-mediated isothermal amplification enabling bright, single-step, closed-tube, and multiplexed detection of RNA viruses

DOE PAGES

Ball, Cameron S.; Light, Yooli K.; Koh, Chung -Yan; ...

2016-03-16

Reverse-transcription-loop-mediated isothermal amplification (RT-LAMP) has frequently been proposed as an enabling technology for simplified diagnostic tests for RNA viruses. However, common detection techniques used for LAMP and RT-LAMP have drawbacks, including poor discrimination capability, inability to multiplex targets, high rates of false positives, and (in some cases) the requirement of opening reaction tubes postamplification. Here, we present a simple technique that allows closed-tube, target-specific detection, based on inclusion of a dye-labeled primer that is incorporated into a target-specific amplicon if the target is present. A short, complementary quencher hybridizes to unincorporated primer upon cooling down at the end of themore » reaction, thereby quenching fluorescence of any unincorporated primer. Our technique, which we term QUASR (for quenching of unincorporated amplification signal reporters, read “quasar”), does not significantly reduce the amplification efficiency or sensitivity of RT-LAMP. Equipped with a simple LED excitation source and a colored plastic gel filter, the naked eye or a camera can easily discriminate between positive and negative QUASR reactions, which produce a difference in signal of approximately 10:1 without background subtraction. We demonstrate that QUASR detection is compatible with complex sample matrices such as human blood, using a novel LAMP primer set for bacteriophage MS2 (a model RNA virus particle). As a result, we demonstrate single-tube duplex detection of West Nile virus (WNV) and chikungunya virus (CHIKV) RNA.« less

Development and Evaluation of a Novel Loop-Mediated Isothermal Amplification Method for Rapid Detection of Severe Acute Respiratory Syndrome Coronavirus

PubMed Central

Thai, Hong Thi Cam; Le, Mai Quynh; Vuong, Cuong Duc; Parida, Manmohan; Minekawa, Harumi; Notomi, Tsugunori; Hasebe, Futoshi; Morita, Kouichi

2004-01-01

The development and evaluation of a one-step single-tube accelerated real-time quantitative reverse transcription (RT) loop-mediated isothermal amplification (LAMP) assay is reported for rapid detection of the severe acute respiratory syndrome coronavirus (SARS-CoV) replicase gene. A total of 49 samples (15 throat washes, 13 throat swabs, and 21 combined throat and nasal swabs) collected from patients admitted to the Hanoi-French and Ninhbinh hospitals in Vietnam during the SARS epidemic were evaluated and compared to conventional RT-PCR. The RT-LAMP assay demonstrated 100-fold-greater sensitivity, with a detection limit of 0.01 PFU. The sensitivity and specificity of RT-LAMP assay for detecting viral RNA in clinical specimens with regard to RT-PCR were 100 and 87%, respectively. The specificity of the RT-LAMP assay was further validated by restriction analysis as well as nucleotide sequencing of the amplified product. The concentration of virus in most of the clinical samples was 0.1 PFU (0.1 to 102 PFU), as determined from the standard curve of SARS RT-LAMP and based on the time of positivity. The assay procedure is quite simple, wherein the amplification is carried out in a single tube under isothermal conditions at 63°C, and the result can be obtained in less than 1 h (as early as 11 min). Thus, the RT-LAMP assay reported here has the advantages of rapid amplification, simple operation, and easy detection and will be useful for rapid and reliable clinical diagnosis of SARS-CoV in developing countries. PMID:15131154

Evidence integration in model-based tree search

PubMed Central

Solway, Alec; Botvinick, Matthew M.

2015-01-01

Research on the dynamics of reward-based, goal-directed decision making has largely focused on simple choice, where participants decide among a set of unitary, mutually exclusive options. Recent work suggests that the deliberation process underlying simple choice can be understood in terms of evidence integration: Noisy evidence in favor of each option accrues over time, until the evidence in favor of one option is significantly greater than the rest. However, real-life decisions often involve not one, but several steps of action, requiring a consideration of cumulative rewards and a sensitivity to recursive decision structure. We present results from two experiments that leveraged techniques previously applied to simple choice to shed light on the deliberation process underlying multistep choice. We interpret the results from these experiments in terms of a new computational model, which extends the evidence accumulation perspective to multiple steps of action. PMID:26324932

A simple heterogeneous one-step assay for screening estrogenic compounds.

PubMed

Huovinen, Tuomas; Rytkönen, Kalle; Lamminmäki, Urpo; Pellinen, Teijo

2013-01-01

Estrogen receptor (ER) modulators are a serious health issue but estrogenic compounds, especially antagonists of ER function, are widely screened for in search of novel therapeutics against hormonal diseases such as the breast cancer. Here we report a novel and a simple bioassay for estrogenic and anti-estrogenic compounds based on ligand-dependent recruitment of ER co-activator steroid receptor co-activator 1 (SRC-1) to purified Renilla luciferase-tagged ERα. In this assay, in vivo-biotinylated (E. coli) SRC-1, purified Renilla luciferase-ERα, and the analyte sample are mixed and incubated for 2 h in a streptavidin-coated microtiter wells, and after one washing step, luminescence is measured with a simple instrument. The assay does not require chemical labeling of the components and shows good sensitivity (25 pM E(2)) and wide dynamic range of more than four orders of magnitude.

Improved Autoassociative Neural Networks

NASA Technical Reports Server (NTRS)

Hand, Charles

2003-01-01

Improved autoassociative neural networks, denoted nexi, have been proposed for use in controlling autonomous robots, including mobile exploratory robots of the biomorphic type. In comparison with conventional autoassociative neural networks, nexi would be more complex but more capable in that they could be trained to do more complex tasks. A nexus would use bit weights and simple arithmetic in a manner that would enable training and operation without a central processing unit, programs, weight registers, or large amounts of memory. Only a relatively small amount of memory (to hold the bit weights) and a simple logic application- specific integrated circuit would be needed. A description of autoassociative neural networks is prerequisite to a meaningful description of a nexus. An autoassociative network is a set of neurons that are completely connected in the sense that each neuron receives input from, and sends output to, all the other neurons. (In some instantiations, a neuron could also send output back to its own input terminal.) The state of a neuron is completely determined by the inner product of its inputs with weights associated with its input channel. Setting the weights sets the behavior of the network. The neurons of an autoassociative network are usually regarded as comprising a row or vector. Time is a quantized phenomenon for most autoassociative networks in the sense that time proceeds in discrete steps. At each time step, the row of neurons forms a pattern: some neurons are firing, some are not. Hence, the current state of an autoassociative network can be described with a single binary vector. As time goes by, the network changes the vector. Autoassociative networks move vectors over hyperspace landscapes of possibilities.

Live-Cell Imaging of Early Steps of Single HIV-1 Infection.

PubMed

Francis, Ashwanth C; Melikyan, Gregory B

2018-05-19

Live-cell imaging of single HIV-1 entry offers a unique opportunity to delineate the spatio-temporal regulation of infection. Novel virus labeling and imaging approaches enable the visualization of key steps of HIV-1 entry leading to nuclear import, integration into the host genome, and viral protein expression. Here, we discuss single virus imaging strategies, focusing on live-cell imaging of single virus fusion and productive uncoating that culminates in HIV-1 infection.

SU-G-BRB-05: Automation of the Photon Dosimetric Quality Assurance Program of a Linear Accelerator

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lebron, S; Lu, B; Yan, G

Purpose: To develop an automated method to calculate a linear accelerator (LINAC) photon radiation field size, flatness, symmetry, output and beam quality in a single delivery for flattened (FF) and flattening-filter-free (FFF) beams using an ionization chamber array. Methods: The proposed method consists of three control points that deliver 30×30, 10×10 and 5×5cm{sup 2} fields (FF or FFF) in a step-and-shoot sequence where the number of monitor units is weighted for each field size. The IC Profiler (Sun Nuclear Inc.) with 5mm detector spacing was used for this study. The corrected counts (CCs) were calculated and the locations of themore » maxima and minima values of the first-order gradient determined data of each sub field. Then, all CCs for each field size are summed in order to obtain the final profiles. For each profile, the radiation field size, symmetry, flatness, output factor and beam quality were calculated. For field size calculation, a parameterized gradient method was used. For method validation, profiles were collected in the detector array both, individually and as part of the step-and-shoot plan, with 9.9cm buildup for FF and FFF beams at 90cm source-to-surface distance. The same data were collected with the device (plus buildup) placed on a movable platform to achieve a 1mm resolution. Results: The differences between the dosimetric quantities calculated from both deliveries, individually and step-and-shoot, were within 0.31±0.20% and 0.04±0.02mm. The differences between the calculated field sizes with 5mm and 1mm resolution were ±0.1mm. Conclusion: The proposed single delivery method proved to be simple and efficient in automating the photon dosimetric monthly and annual quality assurance.« less

Procedure for normalization of cDNA libraries

DOEpatents

Bonaldo, M.D.; Soares, M.B.

1997-12-30

This invention provides a method to normalize a cDNA library constructed in a vector capable of being converted to single-stranded circles and capable of producing complementary nucleic acid molecules to the single-stranded circles comprising: (a) converting the cDNA library in single-stranded circles; (b) generating complementary nucleic acid molecules to the single-stranded circles; (c) hybridizing the single-stranded circles converted in step (a) with complementary nucleic acid molecules of step (b) to produce partial duplexes to an appropriate Cot; (e) separating the unhybridized single-stranded circles from the hybridized single-stranded circles, thereby generating a normalized cDNA library. 1 fig.

Twisting and subunit rotation in single FOF1-ATP synthase

PubMed Central

Sielaff, Hendrik; Börsch, Michael

2013-01-01

FOF1-ATP synthases are ubiquitous proton- or ion-powered membrane enzymes providing ATP for all kinds of cellular processes. The mechanochemistry of catalysis is driven by two rotary nanomotors coupled within the enzyme. Their different step sizes have been observed by single-molecule microscopy including videomicroscopy of fluctuating nanobeads attached to single enzymes and single-molecule Förster resonance energy transfer. Here we review recent developments of approaches to monitor the step size of subunit rotation and the transient elastic energy storage mechanism in single FOF1-ATP synthases. PMID:23267178

A Systems Approach towards an Intelligent and Self-Controlling Platform for Integrated Continuous Reaction Sequences**

PubMed Central

Ingham, Richard J; Battilocchio, Claudio; Fitzpatrick, Daniel E; Sliwinski, Eric; Hawkins, Joel M; Ley, Steven V

2015-01-01

Performing reactions in flow can offer major advantages over batch methods. However, laboratory flow chemistry processes are currently often limited to single steps or short sequences due to the complexity involved with operating a multi-step process. Using new modular components for downstream processing, coupled with control technologies, more advanced multi-step flow sequences can be realized. These tools are applied to the synthesis of 2-aminoadamantane-2-carboxylic acid. A system comprising three chemistry steps and three workup steps was developed, having sufficient autonomy and self-regulation to be managed by a single operator. PMID:25377747

Pretreatment of corn stover using low-moisture anhydrous ammonia (LMAA) process.

PubMed

Yoo, Chang Geun; Nghiem, Nhuan P; Hicks, Kevin B; Kim, Tae Hyun

2011-11-01

A simple pretreatment method using anhydrous ammonia was developed to minimize water and ammonia inputs for cellulosic ethanol production, termed the low moisture anhydrous ammonia (LMAA) pretreatment. In this method, corn stover with 30-70% moisture was contacted with anhydrous ammonia in a reactor under nearly ambient conditions. After the ammoniation step, biomass was subjected to a simple pretreatment step at moderate temperatures (40-120°C) for 48-144 h. Pretreated biomass was saccharified and fermented without an additional washing step. With 3% glucan loading of LMAA-treated corn stover under best treatment conditions (0.1g-ammonia+1.0 g-water per g biomass, 80°C, and 84 h), simultaneous saccharification and cofermentation test resulted in 24.9 g/l (89% of theoretical ethanol yield based on glucan+xylan in corn stover). Copyright © 2011 Elsevier Ltd. All rights reserved.

The Role of the Global SOF Network in a Resource Constrained Environment

DTIC Science & Technology

2013-11-01

backed by actions. Step Two: Matching Skill Sets with Individuals: Who Needs to Know What? This step sounds simple enough in theory . You need to...Training Finally, step five is the recognition that, like physical training and condition- ing, cognitive skills need to be continuously refreshed. These...and maintaining the data needed , and completing and reviewing the collection of information. Send comments regarding this burden estimate or any other