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High Throughput (high + throughput)
Terms modified by High Throughput Selected AbstractsEquilibrium theory analysis of dual reflux PSA for separation of a binary mixtureAICHE JOURNAL, Issue 10 2004Armin D. Ebner A dual reflux (DR) PSA cycle that combines the features of a conventional (stripping reflux) PSA cycle with those of a new enriching reflux PSA cycle is analyzed to show its potential for separating gas mixtures. On the basis of isothermal equilibrium theory applied to linear isotherms, the ultimate separation is carried out where the binary feed is separated into two pure components with 100% recovery of each component. This very idealized analysis reveals that such a separation is possible over a wide range of conditions, even with pressure ratios as low as 1.1. This analysis also reveals that low throughputs and high heavy component recycle ratios are inherently associated with DR PSA cycles, both of which may be detrimental to the process economics. High throughputs and low heavy product recycle ratios are indeed achievable, but only when using low pressure ratios and less selective adsorbents, both counterintuitive results that make sense when considering the perfect separation is always being achieved. Although these trends may not carry over to actual practice, because the model developed here is overly simplified and invalid under certain conditions, this analysis shows that it may indeed be entirely feasible to separate a binary gas mixture into two relatively pure components with very high recoveries using a DR PSA cycle operating with a very low pressure ratio and, hence, expenditure of energy. © 2004 American Institute of Chemical Engineers AIChE J, 50: 2418,2429, 2004 [source] The impact of a supervised injecting facility on ambulance call-outs in Sydney, AustraliaADDICTION, Issue 4 2010Allison M. Salmon ABSTRACT Aims Supervised injecting facilities (SIFs) are effective in reducing the harms associated with injecting drug use among their clientele, but do SIFs ease the burden on ambulance services of attending to overdoses in the community? This study addresses this question, which is yet to be answered, in the growing body of international evidence supporting SIFs efficacy. Design Ecological study of patterns in ambulance attendances at opioid-related overdoses, before and after the opening of a SIF in Sydney, Australia. Setting A SIF opened as a pilot in Sydney's ,red light' district with the aim of accommodating a high throughput of injecting drug users (IDUs) for supervised injecting episodes, recovery and the management of overdoses. Measurements A total of 20 409 ambulance attendances at opioid-related overdoses before and after the opening of the Sydney SIF. Average monthly ambulance attendances at suspected opioid-related overdoses, before (36 months) and after (60 months) the opening of the Sydney Medically Supervised Injecting Centre (MSIC), in the vicinity of the centre and in the rest of New South Wales (NSW). Results The burden on ambulance services of attending to opioid-related overdoses declined significantly in the vicinity of the Sydney SIF after it opened, compared to the rest of NSW. This effect was greatest during operating hours and in the immediate MSIC area, suggesting that SIFs may be most effective in reducing the impact of opioid-related overdose in their immediate vicinity. Conclusions By providing environments in which IDUs receive supervised injection and overdose management and education SIF can reduce the demand for ambulance services, thereby freeing them to attend other medical emergencies within the community. [source] Determination of DNA methylation by COBRA: A comparative study of CGE with LIF detection and conventional gel electrophoresisELECTROPHORESIS, Issue 17 2009Simon Goedecke Abstract DNA methylation as an epigenetic modification of the human genome is under emphatic investigation. Several studies have demonstrated a role of DNA methylation in oncogenesis. In conjunction with histone modifications, DNA methylation may cause the formation of heterochromatin and thus mediate the inactivation of gene transcription. It is important to develop methods that allow for an accurate quantification of the amount of DNA methylation in particular DNA regions, to gain information concerning the threshold of methylation levels necessary for gene inactivation. In this article, a CGE method with on-column LIF detection using SYBR Green is compared with a conventional slab-gel electrophoresis. We thus investigate the validity to analyze DNA methylation in the samples of a combined bisulfite restriction analysis. It is demonstrated that CGE is superior to gel electrophoresis in means of linearity, precision, accuracy, automatization (high throughput), and sample consumption. However, gel electrophoresis is easier to perform (simple devices, no PC usage), and the running costs are comparatively low. A further advantage of CGE is the sparse use of toxic compounds (MeOH and SYBR Green), whereas gel electrophoresis is performed in polyacrylamide gels with ethidium bromide staining. [source] Process Chain for Tailoring the Refractive Index of Thermoplastic Optical Materials using Ceramic NanoparticlesADVANCED ENGINEERING MATERIALS, Issue 6 2005E. Ritzhaupt-Kleissl Interconnections between active optical devices like laser diodes and polymer optical fibres are a crucial factor of optical damping due to coupling losses at the interfaces. Tailoring the refractive index of thermoplastic polymers can diminish these damping losses. The use of modified thermoplastics in an injection moulding process allows a high throughput and therefore a cost effective method for manufacturing passive optical parts with improved properties. [source] Biochemical characterization and inhibitor discovery of shikimate dehydrogenase from Helicobacter pyloriFEBS JOURNAL, Issue 20 2006Cong Han Shikimate dehydrogenase (SDH) is the fourth enzyme involved in the shikimate pathway. It catalyzes the NADPH-dependent reduction of 3-dehydroshikimate to shikimate, and has been developed as a promising target for the discovery of antimicrobial agent. In this report, we identified a new aroE gene encoding SDH from Helicobacter pylori strain SS1. The recombinant H. pylori shikimate dehydrogenase (HpSDH) was cloned, expressed, and purified in Escherichia coli system. The enzymatic characterization of HpSDH demonstrates its activity with kcat of 7.7 s,1 and Km of 0.148 mm toward shikimate, kcat of 7.1 s,1 and Km of 0.182 mm toward NADP, kcat of 5.2 s,1 and Km of 2.9 mm toward NAD. The optimum pH of the enzyme activity is between 8.0 and 9.0, and the optimum temperature is around 60 °C. Using high throughput screening against our laboratory chemical library, five compounds, curcumin (1), 3-(2-naphthyloxy)-4-oxo-2-(trifluoromethyl)-4H -chromen-7-yl 3-chlorobenzoate (2), butyl 2-{[3-(2-naphthyloxy)-4-oxo-2-(trifluoromethyl)-4H -chromen-7-yl]oxy}propanoate (3), 2-({2-[(2-{[2-(2,3-dimethylanilino)-2-oxoethyl]sulfanyl}-1,3-benzothiazol-6-yl)amino]-2-oxoethyl}sulfanyl)- N -(2-naphthyl)acetamide (4), and maesaquinone diacetate (5) were discovered as HpSDH inhibitors with IC50 values of 15.4, 3.9, 13.4, 2.9, and 3.5 µm, respectively. Further investigation indicates that compounds 1, 2, 3, and 5 demonstrate noncompetitive inhibition pattern, and compound 4 displays competitive inhibition pattern with respect to shikimate. Compounds 1, 4, and 5 display noncompetitive inhibition mode, and compounds 2 and 3 show competitive inhibition mode with respect to NADP. Antibacterial assays demonstrate that compounds 1, 2, and 5 can inhibit the growth of H. pylori with MIC of 16, 16, and 32 µg·mL,1, respectively. This current work is expected to favor better understanding the features of SDH and provide useful information for the development of novel antibiotics to treat H. pylori -associated infection. [source] Determination of the mutation spectrum of the EXT1/EXT2 genes in British Caucasian patients with multiple osteochondromas, and exclusion of six candidate genes in EXT negative cases,,HUMAN MUTATION, Issue 11 2006Lorne Lonie Abstract We describe here the spectrum and distribution of mutations in the EXT1 and EXT2 genes in the largest reported British Caucasian multiple osteochondromas (MO) population. Furthermore, we report for the first time the screening of the EXT1 and EXT2 promoters, 5,UTRs, and 3,UTRs, and exclude six potential MO candidate genes in individuals without a detectable mutation within the coding region of EXT1 and EXT2. The coding exons of EXT1 and EXT2 were screened in 72 unrelated probands affected with MO. Forty-six different mutations were identified in 56 probands, of which 29 were novel. Mutation in the EXT1 and EXT2 genes each accounted for 50% of the mutations identified. Of the 72 probands, 42 were of British Caucasian descent, which when added to the 41 British Caucasian families previously reported from our total cohort, gave a total of 83 families. This cohort's proportional frequency for EXT1/EXT2 mutation was 53%/47%. We also validated the technique of high-resolution melting analysis in a blind study using 27 unique EXT1 or EXT2 mutations. This technique was found to be sensitive with a detection rate of 100% regarding heterozygote detection for EXT mutation scanning. Furthermore, this technique has a very high throughput and is very cost-effective. © 2006 Wiley-Liss, Inc. [source] Scanning Probe Parallel Nanolithography with Multiprobe Cantilever Array Fabricated by Bulk Silicon MicromachiningIEEJ TRANSACTIONS ON ELECTRICAL AND ELECTRONIC ENGINEERING, Issue 3 2008Hensy Gandjar Non-member Abstract This work describes a scanning probe parallel nanolithography (SPNL) technique for high throughput in nanometric patterning on single-crystal silicon (SCS) substrates. Two types of multiprobe cantilever arrays used for SPNL were fabricated by conventional micromachining. All the probes mounted on the free end of each cantilever were made of quasitrihedral pyramidal shape composed of (311) and (411) planes using the originally designed mask. Negative and positive types of nanolithography were performed on the basis of field-enhanced anodization and self-assembled monolayer mask techniques, respectively, and they succeeded in drawing a number of nanometric patterns of silicon dioxide (SiO2) on SCS substrates. After anisotropic wet etching of the SCS substrates using the SiO2 films as the mask material, we were also able to fabricate nanowires and nanogrooves. The effects of the applied voltage and scan time of cantilever arrays on wire and groove dimensions were systematically examined by atomic force microscopy (AFM) observations. An optimum condition for the parallel SPNL is proposed on the basis of this research. © 2008 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc. [source] Measured average cell rate-based congestion avoidance schemeINTERNATIONAL JOURNAL OF COMMUNICATION SYSTEMS, Issue 1 2001Hyun M. Choi Abstract Techniques for congestion control of available bit-rate (ABR) traffic in ATM (asynchronous transfer mode) networks remain an important issue. Several congestion control schemes have been proposed to adjust the cell rates of sources with a modified or mean allowed cell rate. To make these schemes work effectively in practice, the modified or mean allowed cell rate must converge under all conditions. However, it is not easy to obtain an accurate value, and an inaccurate value could result in network performance degradation such as severe oscillations and considerable unfairness. Therefore, we propose a measured average cell rate-based congestion avoidance for ABR traffic in ATM networks. The scheme has high throughput and achieves shorter queue lengths without congestion. With measured average cell rate, the scheme provides fast convergence to a start-up virtual connection (VC) and rate of equalization from different initial conditions of the sources. Thus, this scheme provides better fairness among connections. Copyright © 2001 John Wiley & Sons, Ltd. [source] Optimization of multicomponent photopolymer formulations using high-throughput analysis and kinetic modelingAICHE JOURNAL, Issue 5 2010Peter M. Johnson Abstract While high throughput and combinatorial techniques have played an instrumental role in materials development and implementation, numerous problems in materials science and engineering are too complex and necessitate a prohibitive number of experiments, even when considering high throughput and combinatorial approaches, for a comprehensive approach to materials design. Here, we propose a unique combination of high throughput experiments focused on binary formulations that, in combination with advanced modeling, has the potential to facilitate true materials design and optimization in ternary and more complex systems for which experiments are never required. Extensive research on the development of photopolymerizable monomer formulations has produced a vast array of potential monomer/comonomer, initiator and additive combinations. This array dramatically expands the range of material properties that are achievable; however, the vast number of potential formulations has eliminated any possibility of comprehensive materials design or optimization. This limitation is addressed by maximizing the benefits and unique capabilities of high throughput experimentation coupled with predictive models for material behavior and properties. The high throughput experimentation-model combination is useful to collect a limited amount of data from as few as 11 experiments on binary combinations of 10 analyzed monomers, and then use this limited data set to predict and optimize formulation properties in ternary resins that would have necessitated at least 1000 high throughput experiments and several orders of magnitude greater numbers of traditional experiments. A data analysis approach is demonstrated, and the model development and implementation for one model application in which a range of material properties are prescribed, and an optimal formulation that meets those properties is predicted and evaluated. © 2009 American Institute of Chemical Engineers AIChE J, 2010 [source] Millisecond catalytic wall reactors: I. Radiant burnerAICHE JOURNAL, Issue 5 2001J. M. Redenius Short-contact-time reactors have potential for high throughput in reactors much smaller than their traditional counterparts. While they operate adiabatically, heat can be exchanged at short contact time by integrating heat exchange into the reactor. Hot effluent of exothermic reaction systems can be redirected over feed gases to recuperate a portion of the sensible heat. Placing catalyst directly on reactor walls eliminates the resistance to heat transfer in the thermal boundary layer so that heat released by combustion can be effectively coupled to an emitter, such as in a radiant burner. A radiant heater was constructed, operated, and simulated incorporating short contact time, energy recuperation, and a catalytic wall. This burner operated stably for many hours at a firing rate from ,50 to > 160 kW/m2 at a radiant temperature of 950 to 1,150 K at a radiant efficiency of ,60% with a residence time in the reacting zone of ,10 ms. This reactor was modeled using 2-D Navier-Stokes equations including detailed models for chemistry and heat transport. Temperature and compositions predicted agreed well with experimental measurements. [source] Performance of electrospun nanofibers for SPE of drugs from aqueous solutionsJOURNAL OF SEPARATION SCIENCE, JSS, Issue 18 2008Xue-jun Kang Abstract A novel extraction technique was reported. The solid phase material, nanofiber, was prepared by electrospinning using polystyrene. Twenty different drugs (10 ,g/L in water) were extracted using 1 mg of nanofibers within 5 min. The analytes can be desorpted from the fibers with 50 ,L of the methanol and then monitored by LC coupled to a UV detector. Packed-fiber SPE (PFSPE) provide high recoveries (>50%) for some relatively non-polar drugs (log P >1.5) (n -octanol-to-water partition ratio), and relatively low recoveries (9.9,39.8%) for the drugs within the log P window below 1. Experimental optimization of the technique has been carried out using seven representative drugs, edaravone, cinchonine, quinine, voriconazole, chlordiazepoxide, verapamil, and rutonding. Except for edaravone, the maximum yields of seven drugs (0.2 ,g/L) from water samples were approximately 100%, and were 33.7,88.2% from human plasma. The advantageous aspect of the technique encompasses high throughput, high sensitivity, simplicity, low cost, and green chemistry. [source] Analysis of flavor and perfume using an internally cooled coated fiber deviceJOURNAL OF SEPARATION SCIENCE, JSS, Issue 7 2007Yong Chen Abstract A miniaturized internally cooled coated fiber device was applied for the analysis of flavors and fragrances from various matrices. Its integration with a CTC CombiPAL autosampler enabled high throughput for the analysis of analytes in complex matrices that required simultaneous heating of the matrices and cooling of the fiber coating to achieve high extraction efficiency. It was found that up to ten times increase of extraction efficiencies was observed when the device was used to extract flavor compounds in water, even when limited sample temperatures were used to preserve the integrity of target compounds. The extraction of the flavor compounds in water with the device was reproducible, with RSD not larger than 15%. The lower limits of the linear ranges were in the low ppb range, which was about one order of magnitude smaller than those obtained with the commercialized 100 ,m PDMS fibers. Exhaustive extraction of some perfume ingredients from a complex matrix (shampoo) was realized. All achieved recoveries were not less than 80%. The repeatability of the extraction of the perfume compounds from shampoo was better than 10%. The linear ranges were about 1,3000 ,g/g, and the LOD was about 0.2,1 ,g/g. The automated internally cooled coated fiber device was demonstrated to be a powerful sample preparation tool in flavor and fragrance analysis. [source] A twelve-analyzer detector system for high-resolution powder diffractionJOURNAL OF SYNCHROTRON RADIATION, Issue 5 2008Peter L. Lee A dedicated high-resolution high-throughput X-ray powder diffraction beamline has been constructed at the Advanced Photon Source (APS). In order to achieve the goals of both high resolution and high throughput in a powder instrument, a multi-analyzer detector system is required. The design and performance of the 12-analyzer detector system installed on the powder diffractometer at the 11-BM beamline of APS are presented. [source] Bayesian classification of tumours by using gene expression dataJOURNAL OF THE ROYAL STATISTICAL SOCIETY: SERIES B (STATISTICAL METHODOLOGY), Issue 2 2005Bani K. Mallick Summary., Precise classification of tumours is critical for the diagnosis and treatment of cancer. Diagnostic pathology has traditionally relied on macroscopic and microscopic histology and tumour morphology as the basis for the classification of tumours. Current classification frameworks, however, cannot discriminate between tumours with similar histopathologic features, which vary in clinical course and in response to treatment. In recent years, there has been a move towards the use of complementary deoxyribonucleic acid microarrays for the classi-fication of tumours. These high throughput assays provide relative messenger ribonucleic acid expression measurements simultaneously for thousands of genes. A key statistical task is to perform classification via different expression patterns. Gene expression profiles may offer more information than classical morphology and may provide an alternative to classical tumour diagnosis schemes. The paper considers several Bayesian classification methods based on reproducing kernel Hilbert spaces for the analysis of microarray data. We consider the logistic likelihood as well as likelihoods related to support vector machine models. It is shown through simulation and examples that support vector machine models with multiple shrinkage parameters produce fewer misclassification errors than several existing classical methods as well as Bayesian methods based on the logistic likelihood or those involving only one shrinkage parameter. [source] 1H-HRMAS NMR study of smoked Atlantic salmon (Salmo salar)MAGNETIC RESONANCE IN CHEMISTRY, Issue 9 2010David Castejón Abstract High-resolution magic angle spinning (HRMAS) NMR spectroscopic data of smoked Atlantic salmon (Salmo salar) were fully assigned by combination of one- and two-dimensional-HRMAS experiments. Complete representative spectra, obtained after few minutes of analysis time, revealed a large number of minor and major compounds in the sample. The methodology is limited by the low sensitivity of NMR, and therefore HRMAS only enables the determination of the most relevant components. These were fatty acids (FAs), carbohydrates, nucleoside derivatives, osmolytes, amino acids, dipeptides and organic acids. For the first time, spectra were resolved sufficiently to allow semiquantitative determination in intact muscle of the highly polyunsaturated FA 22:6 ,-3. Additionally, the feasibility of 1H-HRMAS NMR metabolite profiling was tested to identify some bioactive compounds during storage. This profiling was carried out by the non-destructive and direct analysis (i.e. without requiring sample preparation and multiple step procedures) of intact salmon muscle. The proposed procedure can be applied to a large number of samples with high throughput due to the short time of analysis and quick evaluation of the data. Copyright © 2010 John Wiley & Sons, Ltd. [source] Biochemical, immunological and clinical characterization of a cross-reactive nonspecific lipid transfer protein 1 from mulberryALLERGY, Issue 5 2010M. A. Ciardiello To cite this article: Ciardiello MA, Palazzo P, Bernardi ML, Carratore V, Giangrieco I, Longo V, Melis M, Tamburrini M, Zennaro D, Mari A, Colombo P. Biochemical, immunological and clinical characterization of a cross-reactive nonspecific lipid transfer protein 1 from mulberry. Allergy 2010; 65: 597,605. Abstract Background:, Mulberry (Morus spp.) is a genus comprising several species of deciduous trees whose fruits are commonly eaten in southern Europe. Subjects with severe systemic reaction have been described. The aim of this study was to isolate the allergens of this species. Methods:, A nonspecific lipid transfer protein 1 (ns-LTP1) was purified from black mulberry by ion exchange and reverse phase high-performance liquid chromatography, and the primary structure was elucidated by direct protein sequencing. Its allergenic activity was evaluated in vivo by skin prick test and in vitro by Western Blot, CD203c basophil activation assay and high throughput multiplex inhibition method on immunosolid-phase allergen chip (ISAC). Results:, Mulberry ns-LTP (Mor n 3) comprises 91 amino acids producing a molecular mass of 9246 Da. This protein shows high sequence identity with several allergenic ns-LTP1. Immunoblot analysis and CD203c activation assay demonstrated its allergenic activity in symptomatic subjects and in ns-LTP allergic patients who are not mulberry consumers. Immunological co-recognition was studied in vivo on a selected group of well-characterized ns-LTP allergic patients showing a high percentage of nMor n 3+ subjects (88.46%) even in patients who have never eaten mulberry before. IgE inhibition on ISAC micro-array demonstrated an almost complete cross-reactivity to nArt v 3, rCor a 8 and a very high percentage of inhibition to nPru p 3. Conclusions:, Mor n 3 is the first allergen isolated in black mulberry and immunologically characterized. It displayed allergenic activity among symptomatic and nonconsumer patients and a pattern of cross-reactivity to other plant-derived LTPs. [source] A fast and inexpensive DNA extraction/purification protocol for brown macroalgaeMOLECULAR ECOLOGY RESOURCES, Issue 2 2007GALICE HOARAU Abstract Here we describe a rapid method for extracting DNA from dried brown algae material using a microtitre plate system in conjunction with a milling instrument. The method allows the preparation of nuclear and organelle DNA of quality suitable for polymerase chain reaction amplification. It combines high throughput with low cost per sample: DNA from 192 samples can be extracted in c. 3 h for < ,0.40 per sample, nearly tenfold cheaper than commercially available kits. Furthermore, by using microtitre plates, efficient storage and downstream processing is facilitated. [source] Genotyping of pantophysin I (Pan I) of Atlantic cod (Gadus morhua L.) by allele-specific PCRMOLECULAR ECOLOGY RESOURCES, Issue 1 2006JŘRGEN STENVIK Abstract The two main allelic variants of the Atlantic cod (Gadus morhua L.) pantophysin I (Pan I) locus have different frequencies within different cod stocks. The Dra I polymorphism which distinguishes the two alleles can thus be used for discrimination of coastal and offshore cod populations. We present a new method for Pan I genotyping using fluorescent allele-specific duplex polymerase chain reaction (PCR). This method is more rapid, reliable and cost-effective than the previously published method and it is not affected by DNA source and quality. This improvement is important for studies demanding high throughput and accuracy of Pan I genotyping [source] Development of biosensor based on imaging ellipsometryPHYSICA STATUS SOLIDI (A) APPLICATIONS AND MATERIALS SCIENCE, Issue 4 2008G. Jin Abstract The concept of biosensor based on imaging ellipsometry was proposed ten years ago. Its principle and the methodology as well as some solutions to problems which have to be faced during the development are mentioned. Its properties of phase sensitive, high throughput and fast sampling, as well as label-free, sensitivity better than 1 ng/ml for Immunoglobulin G, and real-time analysis for protein interaction process, etc. provide a potential for applications in biomedicine field. The recent biosensing development with total internal reflection imaging ellipsometry is presented also. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Quantifying the three main components of salinity tolerance in cerealsPLANT CELL & ENVIRONMENT, Issue 3 2009KARTHIKA RAJENDRAN ABSTRACT Salinity stress is a major factor inhibiting cereal yield throughout the world. Tolerance to salinity stress can be considered to contain three main components: Na+ exclusion, tolerance to Na+ in the tissues and osmotic tolerance. To date, most experimental work on salinity tolerance in cereals has focused on Na+ exclusion due in part to its ease of measurement. It has become apparent, however, that Na+ exclusion is not the sole mechanism for salinity tolerance in cereals, and research needs to expand to study osmotic tolerance and tissue tolerance. Here, we develop assays for high throughput quantification of Na+ exclusion, Na+ tissue tolerance and osmotic tolerance in 12 Triticum monococcum accessions, mainly using commercially available image capture and analysis equipment. We show that different lines use different combinations of the three tolerance mechanisms to increase their total salinity tolerance, with a positive correlation observed between a plant's total salinity tolerance and the sum of its proficiency in Na+ exclusion, osmotic tolerance and tissue tolerance. The assays developed in this study can be easily adapted for other cereals and used in high throughput, forward genetic experiments to elucidate the molecular basis of these components of salinity tolerance. [source] A high throughput membrane BIO-PCR technique for ultra-sensitive detection of Pseudomonas syringae pv. phaseolicolaPLANT PATHOLOGY, Issue 1 2007N. W. Schaad Molecular-based methods such as PCR have greatly improved detection of bacteria in environmental samples. However, the sensitivity of PCR is not high when compared to agar plating assays, and inhibitors from plants are often a problem. Pre-enriching bacteria on agar media (BIO-PCR) can increase the sensitivity of PCR by more than 100% and reduce the effects of inhibitors. To further increase the sensitivity and also reduce the labour needed for BIO-PCR, a high throughput 96-well membrane BIO-PCR technique is described for ultra-sensitive detection of Pseudomonas syringae pv. phaseolicola (PSP) (syn. P. phaseolicola) in washings of seeds and leaves of Phaseolus vulgaris, using available classical PCR primers and newly designed real-time primers and probe. The primers and probe, designed from a tox-argK chromosomal cluster of the PSP-specific phaseolotoxin gene, were confirmed to be specific to PSP. Samples (1·2 mL) were filtered under vacuum in 96-well membrane plates. After incubating on soft agar medium for 48,52 h, each well is washed with 200 µL of sterile water and used immediately for nested (two-step) PCR or real-time PCR or stored at ,20°C. Results of assaying spiked seed washings showed that classical PCR was unable to detect PSP at mean concentrations of 40 colony forming units (cfu) mL,1. BIO-PCR detected PSP in five out of six samples at 40 mean cfu mL,1 but none at mean concentrations of 4·2 and 0·4 mean cfu mL,1. In contrast, membrane BIO-PCR detected the bacterium in all six samples tested containing as few as 0·4 mean cfu mL,1. The sensitivity of detection from leaf washings was lower but the results were similar, classical and BIO-PCR were negative from all three levels of inoculum while membrane BIO-PCR detected three out of three samples at 80 mean cfu mL,1 and one out of three at 40 mean cfu mL,1. [source] Thin film solar modules: the low cost, high throughput and versatile alternative to Si wafersPROGRESS IN PHOTOVOLTAICS: RESEARCH & APPLICATIONS, Issue 5 2006S. Hegedus Abstract Thin film solar cells (TFSC) have passed adolescence and are ready to make a substantial contribution to the world's electricity generation. They can have advantages over c-Si solar modules in ease of large area, lower cost manufacturing and in several types of applications. Factors which limit TFSC module performance relative to champion cell performance are discussed along with the importance of increased throughput and yield. The consensus of several studies is that all TFSC can achieve costs below 1,$/W if manufactured at sufficiently large scale >100,MW using parallel lines of cloned equipment with high material utilization and spray-on encapsulants. There is significant new commercial interest in TFSC from small investors and large corporations, validating the thin film approach. Unique characteristics are discussed which give TFSC an advantage over c-Si in two specific markets: small rural solar home systems and building integrated photovoltaic installations. TFSC have outperformed c-Si in annual energy production (kWhrs/kW), have demonstrated outdoor durability comparable to c-Si and are being used in MW scale installations worldwide. The merits of the thin film approach cannot be judged on the basis of efficiency alone but must also account for module performance and potential for low cost. TFSC advocates should promote their unique virtues compared to c-Si: lower cost, higher kWhr/kW output, higher battery charging current, attractive visual appearance, flexible substrates, long-term stability comparable to c-Si, and multiple pathways for deposition with room for innovation and evolutionary improvement. There is a huge market for TFSC even at today's efficiency if costs can be reduced. A brief window of opportunity exists for TFSC over the next few years due the Si shortage. The demonstrated capabilities and advantages of TFSC must be proclaimed more persistently to funding decision-makers and customers without minimizing the remaining challenges. Copyright © 2006 John Wiley & Sons, Ltd. [source] High-quality surface passivation of silicon solar cells in an industrial-type inline plasma silicon nitride deposition systemPROGRESS IN PHOTOVOLTAICS: RESEARCH & APPLICATIONS, Issue 1 2004Jens D. Moschner Abstract We have studied the surface passivation of silicon by deposition of silicon nitride (SiN) in an industrial-type inline plasma-enhanced chemical vapor deposition (PECVD) reactor designed for the continuous coating of silicon solar cells with high throughput. An optimization study for the passivation of low-resistivity p -type silicon has been performed exploring the dependence of the film quality on key deposition parameters of the system. With the optimized films, excellent passivation properties have been obtained, both on undiffused p -type silicon and on phosphorus-diffused n+ emitters. Using a simple design, solar cells with conversion efficiencies above 20% have been fabricated to prove the efficacy of the inline PECVD SiN. The passivation properties of the films are on a par with those of high-quality films prepared in small-area laboratory PECVD reactors. Copyright © 2004 John Wiley & Sons, Ltd. [source] Serum protein profiling by solid phase extraction and mass spectrometry: A future diagnostics tool?PROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 6 2009Anne K. Callesen Abstract Serum protein profiling by MS is a promising method for early detection of disease. Important characteristics for serum protein profiling are preanalytical factors, analytical reproducibility and high throughput. Problems related to preanalytical factors can be overcome by using standardized and rigorous sample collection and sample handling protocols. The sensitivity of the MS analysis relies on the quality of the sample; consequently, the blood sample preparation step is crucial to obtain pure and concentrated samples and enrichment of the proteins and peptides of interest. This review focuses on the serum sample preparation step prior to protein profiling by MALDI MS analysis, with particular focus on various SPE methods. The application of SPE techniques with different chromatographic properties such as RP, ion exchange, or affinity binding to isolate specific subsets of molecules (subproteomes) is advantageous for increasing resolution and sensitivity in the subsequent MS analysis. In addition, several of the SPE sample preparation methods are simple and scalable and have proven easy to automate for higher reproducibility and throughput, which is important in a clinical proteomics setting. [source] A novel approach and protocol for discovering extremely low-abundance proteins in serumPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 17 2006Yoshinori Tanaka Abstract The proteomic analysis of serum (plasma) has been a major approach to determining biomarkers essential for early disease diagnoses and drug discoveries. The determination of these biomarkers, however, is analytically challenging since the dynamic concentration range of serum proteins/peptides is extremely wide (more than 10,orders of magnitude). Thus, the reduction in sample complexity prior to proteomic analyses is essential, particularly in analyzing low-abundance protein biomarkers. Here, we demonstrate a novel approach to the proteomic analyses of human serum that uses an originally developed serum protein separation device and a sequentially linked 3-D-LC-MS/MS system. Our hollow-fiber-membrane-based serum pretreatment device can efficiently deplete high-molecular weight proteins and concentrate low-molecular weight proteins/peptides automatically within 1,h. Four independent analyses of healthy human sera pretreated using this unique device, followed by the 3-D-LC-MS/MS successfully produced 12,000,13,000 MS/MS spectra and hit around 1800,proteins (>95% reliability) and 2300,proteins (>80% reliability). We believe that the unique serum pretreatment device and proteomic analysis protocol reported here could be a powerful tool for searching physiological biomarkers by its high throughput (3.7,days per one sample analysis) and high performance of finding low abundant proteins from serum or plasma samples. [source] Rapid identification of differentiation markers from whole epithelial cells by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry and statistical analysisRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 8 2008Laure F. Marvin-Guy Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS) was applied to identify markers for cellular differentiation. The differentiation of a human colon epithelial carcinoma T84 cell line was monitored over a period of 28 days by transepithelial electrical resistance (TER) measurements, alkaline phosphatase (AP) assay, and MALDI-TOF mass spectral fingerprints combined with statistical analysis. MALDI-MS generated specific mass spectral fingerprints characteristic of cell differentiation. Twenty-two ions were selected as diagnostic signals of fully differentiated T84 cells. Ten protein ion signals, detected by MALDI-MS and validated by statistical analysis, were proposed as T84 cell differentiation markers. Among these signals, ubiquitin was identified as a T84 cell differentiation marker by nanospray liquid chromatography/tandem mass spectrometry (nanoLC/MS/MS). Moreover, depending on the concentration of the cells seeded on the growth support, it was possible to predict the timing of the exponential phase and of cellular differentiation by MALDI-MS-derived marker ions. MALDI-TOFMS was compared to other methods for the determination of cellular differentiation: TER measurements are rapid but yield limited information as to the cellular differentiation state. AP assays are more specific for the differentiation state but take more time. By contrast, MALDI-MS has been found to be a fast, sensitive and precise method for cell differentiation assessment and provides the opportunity for multiplexing and high throughput. Moreover, the consumable costs per assay are very low. Copyright © 2008 John Wiley & Sons, Ltd. [source] Use of flow injection atmospheric pressure photoionization quadrupole time-of-flight mass spectrometry for fast olive oil fingerprintingRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 8 2006J. L. Gómez-Ariza The recently introduced technique of an atmospheric pressure photoionization (APPI) source coupled to quadrupole time-of-flight mass spectrometry (QqTOFMS) has been applied to fast olive oil fingerprinting on the basis of the accurate mass measurements obtained with this instrumentation. The key compounds can be characterized as [M+H]+ (produced by proton transfer) or as [M]+. (by charge transfer) ions in the mass spectra. [M+H]+ ions, however, show higher abundance, especially for triacylglycerols. Other ions present in APPI-MS are the acylium ion [RiCO]+ and [RiCOH2O]+. This latter ion is absent in the electrospray ionization (ESI)-MS spectra, and this represents valuable complementary information. Several critical parameters in the APPI source were optimized such as LC eluent composition, ion spray voltage and, especially, declustering potential. APPI-QqTOFMS allows easy discrimination among different edible oils: olive, extra virgin olive, olive-pomace, hazelnut, sunflower, corn and several mixed oils, with high throughput (approximately 1,min per sample). Cluster analysis was applied to obtain the best experimental conditions for oil discrimination on the basis of declustering potential. Principal components analyses of these APPI-MS spectra show that the approach can be used for studies of olive oil adulteration with other oils, even in the case of hazelnut oil that exhibits a high chemical similarity with olive oil. Copyright © 2006 John Wiley & Sons, Ltd. [source] Development of a liquid chromatographic time-of-flight mass spectrometric method for the determination of unlabelled and deuterium-labelled , -tocopherol in blood componentsRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 24 2003Wendy L. Hall A method is described for the analysis of deuterated and undeuterated , -tocopherol in blood components using liquid chromatography coupled to an orthogonal acceleration time-of-flight (TOF) mass spectrometer. Optimal ionisation conditions for undeuterated (d0) and tri- and hexadeuterated (d3 or d6) , -tocopherol standards were found with negative ion mode electrospray ionisation. Each species produced an isotopically resolved single ion of exact mass. Calibration curves of pure standards were linear in the range tested (0,1.5,,M, 0,15,pmol injected). For quantification of d0 and d6 in blood components following a standard solvent extraction, a stable-isotope-labelled internal standard (d3- , -tocopherol) was employed. To counter matrix ion suppression effects, standard response curves were generated following identical solvent extraction procedures to those of the samples. Within-day and between-day precision were determined for quantification of d0- and d6-labelled , -tocopherol in each blood component and both averaged 3,10%. Accuracy was assessed by comparison with a standard high-performance liquid chromatography (HPLC) method, achieving good correlation (r2,=,0.94), and by spiking with known concentrations of , -tocopherol (98% accuracy). Limits of detection and quantification were determined to be 5 and 50,fmol injected, respectively. The assay was used to measure the appearance and disappearance of deuterium-labelled , -tocopherol in human blood components following deuterium-labelled (d6) RRR - , -tocopheryl acetate ingestion. The new LC/TOFMS method was found to be sensitive, required small sample volumes, was reproducible and robust, and was capable of high throughput when large numbers of samples were generated. Copyright © 2003 John Wiley & Sons, Ltd. [source] Phenotyping approaches for physiological breeding and gene discovery in wheatANNALS OF APPLIED BIOLOGY, Issue 3 2009M. Reynolds Abstract Conceptual models of drought-adaptive traits have been used in breeding to accumulate complementary physiological traits (PT) in selected progeny, resulting in distribution of advanced lines to rain-fed environments worldwide by the International Maize and Wheat Improvement Center (CIMMYT). Key steps in PT breeding at CIMMYT include characterisation of crossing block lines for stress adaptive mechanisms, strategic crossing among parents that encompass as many target traits as possible and early generation selection (EGS) of bulks for canopy temperature (CT). The approach has been successful using both elite × elite crosses as well as three way crosses involving stress adapted landraces. Other EGS techniques that are amenable to high throughput include measurement of spectral reflectance indices and stomatal aperture-related traits. Their genetic- and cost-effectiveness are supported by realisation of genetic yield gains in response to trait selection, and by economic analysis, respectively. Continual reselection within restricted gene pools is likely to lead to diminishing returns, however, exotic parents can be used to introduce new allelic diversity. Examples include landraces from the primary gene pool, and products of inter-specific hybridisation with the secondary gene pool consisting of closely related wheat genomes. Both approaches have been successful in introducing stress-adaptive traits. The main problem with knowing which genetic resource to use in wide-crossing is the uncertainty with which phenotypic expression can be extrapolated from one genome/genepool to another because of their unimproved or undomesticated genetic backgrounds. Nonetheless, their PT expression can be measured and used as a basis for investing in crossing or wide crossing. Discovering the genetic basis of PT is highly complex because putative QTLs may interact with environment and genetic background, including genes of major effect. Detection of QTLs was improved in mapping populations where flowering time was controlled, while new mapping populations have been designed by screening potential parents that do not contrast in the Rht, Ppd and Vrn alleles. Association genetics mapping is another approach that can be employed for gene discovery using exclusively agronomically improved material, thereby minimising the probability of identifying yield QTLs whose alleles have been already improved by conventional breeding. [source] Protein crystallization for genomics: towards high-throughput optimization techniquesACTA CRYSTALLOGRAPHICA SECTION D, Issue 6-2 2002Naomi E. Chayen Protein crystallization has gained a new strategic and commercial relevance in the next phase of the genome projects, in which X-ray crystallography will play a major role. Considerable advances have been made in the automation of protein preparation and also in the X-ray analysis and bioinformatics stages once diffraction-quality crystals are available. These advances have not yet been matched by equally good methods for the crystallization process itself. In the area of crystallization, the main effort and resources are currently being invested into the automation of screening procedures to identify potential crystallization conditions. However, in spite of the ability to generate numerous trials, so far only a small percentage of the proteins produced have led to structure determinations. This is because screening in itself is not usually enough; it has to be complemented by an equally important procedure in crystal production, namely crystal optimization. In the rush towards structural genomics, optimization techniques have been somewhat neglected, mainly because it was hoped that large-scale screening alone would produce the desired results. In addition, optimization has relied on particular individual methods that are often difficult to automate and to adapt to high throughput. This article addresses a major gap in the field of structural genomics by describing practical ways of automating individual optimization methods in order to adapt them to high-throughput techniques. [source] | ||||||||||||||||||||||