A polymerase chain reaction-based method for isolating clones from a complimentary DNA library in sheep

The sheep (Ovis aries) is favored by many musculoskeletal tissue engineering groups as a large animal model because of its docile temperament and ease of husbandry. The size and weight of sheep are comparable to humans, which allows for the use of implants and fixation devices used in human clinical practice. The construction of a complimentary DNA (cDNA) library can capture the expression of genes in both a tissue- and time-specific manner. cDNA libraries have been a consistent source of gene discovery ever since the technology became commonplace more than three decades ago. Here, we describe the construction of a cDNA library using cells derived from sheep bones based on the pBluescript cDNA kit. Thirty clones were picked at random and sequenced. This led to the identification of a novel gene, C12orf29, which our initial experiments indicate is involved in skeletal biology. We also describe a polymerase chain reaction-based cDNA clone isolation method that allows the isolation of genes of interest from a cDNA library pool. The techniques outlined here can be applied in-house by smaller tissue engineering groups to generate tools for biomolecular research for large preclinical animal studies and highlights the power of standard cDNA library protocols to uncover novel genes.

Isolating CP-violating gamma ZZ coupling in e(+)e(-) -> gamma Z with transverse beam polarizations

We revisit the process e(+)e(-) -> gamma Z at the ILC with transverse beam polarization in the presence of anomalous CP- violating gamma ZZ coupling lambda(1) and gamma gamma Z coupling lambda(2). We point out that if the final- state spins are resolved, then it becomes possible to fingerprint the anomalous coupling Re lambda(1). 90% confidence level limit on Re lambda(1) achievable at ILC with center- of- mass energy of 500 GeVor 800 GeV with realistic initial beam polarization and integrated luminosity is of the order of few times of 10(-2) when the helicity of Z is used and 10(-3) when the helicity of gamma is used. The resulting corrections at quadratic order to the cross section and its influence on these limits are also evaluated and are shown to be small. The benefits of such polarization programmes at the ILC are compared and contrasted for the process at hand. We also discuss possible methods by which one can isolate events with a definite helicity for one of the final- state particles.

Isolating the impacts of land use and climate change on streamflow

Quantifying the isolated and integrated impacts of land use (LU) and climate change on streamflow is challenging as well as crucial to optimally manage water resources in river basins. This paper presents a simple hydrologic modeling-based approach to segregate the impacts of land use and climate change on the streamflow of a river basin. The upper Ganga basin (UGB) in India is selected as the case study to carry out the analysis. Streamflow in the river basin is modeled using a calibrated variable infiltration capacity (VIC) hydrologic model. The approach involves development of three scenarios to understand the influence of land use and climate on streamflow. The first scenario assesses the sensitivity of streamflow to land use changes under invariant climate. The second scenario determines the change in streamflow due to change in climate assuming constant land use. The third scenario estimates the combined effect of changing land use and climate over the streamflow of the basin. Based on the results obtained from the three scenarios, quantification of isolated impacts of land use and climate change on streamflow is addressed. Future projections of climate are obtained from dynamically downscaled simulations of six general circulation models (GCMs) available from the Coordinated Regional Downscaling Experiment (CORDEX) project. Uncertainties associated with the GCMs and emission scenarios are quantified in the analysis. Results for the case study indicate that streamflow is highly sensitive to change in urban areas and moderately sensitive to change in cropland areas. However, variations in streamflow generally reproduce the variations in precipitation. The combined effect of land use and climate on streamflow is observed to be more pronounced compared to their individual impacts in the basin. It is observed from the isolated effects of land use and climate change that climate has a more dominant impact on streamflow in the region. The approach proposed in this paper is applicable to any river basin to isolate the impacts of land use change and climate change on the streamflow.

Isolating single primary rat hippocampal neurons and astrocytes on ultra-thin patterned parylene-C/silicon dioxide substrates

We report here the patterning of primary rat neurons and astrocytes from the postnatal hippocampus on ultra-thin parylene-C deposited on a silicon dioxide substrate, following observations of neuronal, astrocytic and nuclear coverage on strips of different lengths, widths and thicknesses. Neuronal and glial growth was characterized 'on', 'adjacent to' and 'away from' the parylene strips. In addition, the article reports how the same material combination can be used to isolate single cells along thin tracks of parylene-C. This is demonstrated with a series of high magnification images of the experimental observations for varying parylene strip widths and thicknesses. Thus, the findings demonstrate the possibility to culture cells on ultra-thin layers of parylene-C and localize single cells on thin strips. Such work is of interest and significance to the Neuroengineering and Multi-Electrode Array (MEA) communities, as it provides an alternative insulating material in the fabrication of embedded micro-electrodes, which can be used to facilitate single cell stimulation and recording in capacitive coupling mode. © 2010 Elsevier Ltd.

Isolating and recognizing crudeoiklegrading [sic] bacteria of Queshm Island shores for biodegradation of crude oil pollutions

This research was carried out for recognizing Natural Flora Bacteria of oil pollution in the coasts of Queshm island. In The First steps, The coasts of this Island were scrutinized as a Field of research and For knowing whether oil stains exist or not. It gets obvious That southern coasts of Queshm have got oil pollution which is created by oil tankers which carry oil of Iran continental shelf. Them oil stains were sampled from to certain stations. In The First step, primary isolation of exisiting bacteria in every oil sample was done and then purification of each bacterium was carried out. Then each purified bacterium that has got strong, recognized, typic growth was enriched oil sample of T5 station. And Bacterium C4 (gram—negative coccobacillus) was chosen as the second priority From oil sample of TA station and Bacterium B1 (gram—positive coccus) was chosen as The third priority From oil sample of TI station. All The above mentioned bacteria were biochemically, physiologically and morphologically experimented For specking The species. According To The tests done and comparing with The tests done and comparing with the reference Berge y' s, bacterium A5 Pelongs to the species pseudomonas sp and becterium C4 belongs to the species Aeromonas sp and bacterium BI belongs to The species micrococcus sp. In The Last stage, bacterium with The First priority (TA5 pseudomonas sp) was used in the planned microcosm. The sake of optimum and adapting to Laboratory conditions Each enriched and purified bacterium was given a code for station and a code For itself . Then This bacterium was studied and it was proved that it has potentiality For using oil as a source of carbon. From oil samples of 10 stations, 30 various Colonies of bacterium were Isolated, of which 20 bacteria had the highest potentiality of growth. And the other bacteria that has no typic growth were omitted From being studied. Since all of These 20 bacterium are able to use oil, a bacterium with maximum rate of growth in the presence of crude oil and Lack of other hydrocarbonic sources and with The code A5 ( gram — negative Bacillus ) was chosen as First priority From The mentioned microcosm contains sea water , suspension oil degrading bacterium , crude oil, azote and various concentrations of carbon and Incubated in 30°` and shook 150 PRA1 According to the results , index oil degrading bacterium (pseudomonas sp) belongs oil sample of T5 stations (east of sheeb draz Gulf) which growth best and have the potentiality of degrading oil in 25 glli malas and 50 glli cheese water and with 5 gill urea .

Isolating shallow foundations from seismic loading

Most modern design codes do not allow for movement between a shallow foundation and the underlying soil during seismic loading. Consequently, the full magnitude of seismic energy is transmitted from the soil to the foundation during an earthquake. This energy either has to be dissipated before reaching the superstructure via engineering solutions such as base isolation systems, or the structure itself must withstand the full impact of the earthquake resulting in high material usage and expensive design. However, the inherent hysteric behaviour of soil can be used to isolate a foundation from the underlying soil. As part of a study into the soil-structure-interaction of shallow foundations, methods to optimise foundation isolation were investigated. In this paper the results from centrifuge tests investigating two of these methods are compared to results when no special foundation layout was implemented and the impact of the proposed isolation methods is discussed. © 2010 Taylor & Francis Group, London.

Influence of thermal isolating grooves on the performance of the Mach-Zehnder interferometer-type thermo-optic variable optical attenuator

Two types of silicon-on-insulator thermo-optic variable optical attenuators (VOAs) based on a Mach-Zehnder interferometer and a multimode-interference coupler are fabricated, one with thermal isolating grooves to improve heating efficiency and the other without Comparison of optical and electrical properties, such as insertion losses, the maximum attenuation levels and the corresponding power consumptions, and the response times, is carried out between the two types of VOAs. The comparison results Indicate that use of thermal isolating grooves leads to better values for most characteristics and is an effective way to improve the performance of Mach-Zehnder interferometer-type thermo-optic devices. (c) 2005 Society of Photo-Optical Instrumentation Engineers.

The effect of buried AlxGa1-xN isolating layers on the transport properties of GaN deposited on sapphire substrate by molecular beam epitaxy using NH3

It is believed that the highly dislocated region near the GaN/sapphire interface is a degenerate layer. In this paper a direct evidence for such a proposal is presented. By inserting a buried AlxGa1-xN (x > 0.5) isolating layer to separate the interface region from the bulk region, the background electron concentration can be significantly reduced, while care must be taken to guarantee that there is no degrading of Hall mobility when choosing the thickness of the isolating layer. (C) 1998 Elsevier Science B.V. All rights reserved.

Design considerations for a silicon-based p-i-n phase modulator in a double ridge waveguide with side isolating grooves

We present detail design considerations and simulation results of a forward biased carrier injection p-i-n modulator integrated on SOI rib waveguides. To minimize the free carrier absorption loss while keeping the comparatively small lateral dimensions of the modulator as required for high speed operation, we proposed two structural improvements, namely the double ridge (terrace ridge) structure and the isolating grooves at both sides of the double ridge. With improved carrier injection and optical confinement structure, the simulated modulator response time is in sub-ns range and absorption loss is minimized.

One-pot two-step mechanochemical synthesis: ligand and complex preparation without isolating intermediates

Although the use of ball milling to induce reactions between solids (mechanochemical synthesis) can provide lower-waste routes to chemical products by avoiding solvent during the reaction, there are further potential advantages in using one-pot multistep syntheses to avoid the use of bulk solvents for the purification of intermediates. We report here two-step syntheses involving formation of salen-type ligands from diamines and hydroxyaldehydes followed directly by reactions with metal salts to provide the corresponding metal complexes. Five salen-type ligands 2,2'-[1,2-ethanediylbis[(E)-nitrilomethylidyne]] bisphenol, ` salenH2', 1; 2,2'-[(+/-)-1,2-cyclohexanediylbis-[(E)-nitrilomethylidyne]] bis-phenol, 2; 2,2'-[1,2-phenylenebis( nitrilomethylidyne)]-bis-phenol, ` salphenH2' 3; 2-[[(2-aminophenyl) imino] methyl]-phenol, 4; 2,2'-[(+/-)-1,2-cyclohexanediylbis[(E)-nitrilomethylidyne]]-bis[4,6-bis(1,1-dimethylethyl)]-phenol, ` Jacobsen ligand', 5) were found to form readily in a shaker-type ball mill at 0.5 to 3 g scale from their corresponding diamine and aldehyde precursors. Although in some cases both starting materials were liquids, ball milling was still necessary to drive those reactions to completion because precipitation of the product and or intermediates rapidly gave in thick pastes which could not be stirred conventionally. The only ligand which required the addition of solvent was the Jacobsen ligand 5 which required 1.75 mol equivalents of methanol to go to completion. Ligands 1-5 were thus obtained directly in 30-60 minutes in their hydrated forms, due to the presence of water by-product, as free-flowing yellow powders which could be dried by heating to give analytically pure products. The one-armed salphen ligand 4 could also be obtained selectively by changing the reaction stoichiometry to 1 : 1. SalenH(2) 1 was explored for the onepot two-step synthesis of metal complexes. In particular, after in situ formation of the ligand by ball milling, metal salts (ZnO, Ni(OAc)2 center dot 4H(2)O or Cu(OAc)(2)center dot H2O) were added directly to the jar and milling continued for a further 30 minutes. Small amounts of methanol (0.4-1.1 mol equivalents) were needed for these reactions to run to completion. The corresponding metal complexes [M(salen)] (M = Zn, 6; Ni, 7; or Cu, 8) were thus obtained quantitatively after 30 minutes in hydrated form, and could be heated briefly to give analytically pure dehydrated products. The all-at-once ` tandem' synthesis of [Zn(salen)] 6 was also explored by milling ZnO, ethylene diamine and salicylaldehyde together in the appropriate mole ratio for 60 minutes. This approach also gave the target complex selectively with no solvent needing to be added. Overall, these syntheses were found to be highly efficient in terms of time and the in avoidance of bulk solvent both during the reaction and for the isolation of intermediates. The work demonstrates the applicability of mechanochemical synthesis to one-pot multi-step strategies.

Isolating Informational Masking in Both Pure and Complex Tone Sequences

Objectives: Interference between a target and simultaneous maskers occurs both at the cochlear level through energetic masking and more centrally through informational masking (IM). Hence, quantifying the amount of IM requires a strict control of the energetic component. Presenting target and maskers on different sides (i.e., dichotically) reduces energetic masking but provides listeners with important lateralization cues that also drastically reduce IM. The main purpose of this study (Experiment 1) was to evaluate a "switch" manipulation aiming at restoring most of the IM despite dichotic listening. Experiment 2 was designed to investigate the source of the difficulty induced by this switching dichotic condition.

Design: In Experiment 1, the authors presented 60 normal-hearing young adults with a detection task in which a regularly repeating target was embedded in a randomly varying background masker. The authors evaluated spatial masking release induced by three different dichotic listening conditions in comparison with a diotic baseline. Dichotic stimuli were presented in either a nonswitching or a switching condition. In the latter case, the presentation sides of dichotic target and maskers alternated several times throughout 10 sec sequences. The impact of the number of switches on IM was investigated parametrically, with both pure and complex tone sequences. In Experiment 2, the authors compared performance of 13 young, normal-hearing listeners in a monotic and dichotic version of the rapidly switching condition, using pure-tone sequences.

Results: When target and maskers switched rapidly within sequences, IM was significantly stronger than in nonswitching dichotic sequences and was comparable with the masking effect induced by diotic sequences. Furthermore, Experiment 2 suggests that rapidly switching target and maskers prevent listeners from relying on lateralization cues inherent to the dichotic condition, hence preserving important amounts of IM.

Conclusions: This paradigm thus provides an original tool to isolate IM in signal and maskers having overlapping spectra.

Isolating the signal of ocean global warming

Identifying the signature of global warming in the world's oceans is challenging because low frequency circulation changes can dominate local temperature changes. The IPCC fourth assessment reported an average ocean heating rate of 0.21 ± 0.04 Wm−2 over the period 1961–2003, with considerable spatial, interannual and inter-decadal variability. We present a new analysis of millions of ocean temperature profiles designed to filter out local dynamical changes to give a more consistent view of the underlying warming. Time series of temperature anomaly for all waters warmer than 14°C show large reductions in interannual to inter-decadal variability and a more spatially uniform upper ocean warming trend (0.12 Wm−2 on average) than previous results. This new measure of ocean warming is also more robust to some sources of error in the ocean observing system. Our new analysis provides a useful addition for evaluation of coupled climate models, to the traditional fixed depth analyses.

Isolating the 'farmer' effect as a component of the advantage of growing genetically modified varieties in developing countries: a Bt cotton case study from Jalgaon, India

The present paper explores the 'farmer' effect in economic advantages often claimed for Bt cotton varieties (those with the endotoxin gene from Bacillus thuringiensis conferring resistance to some insect pests) compared to non-Bt varieties. Critics claim that much of the yield advantage of Bt cotton could be due to the fact that farmers adopting the technology are in a better position to provide inputs and management and so much of any claimed Bt advantage is an artefact rather than reflecting a real advantage of the variety per se. The present paper provides an in-depth analysis of 63 non-adopting and 94 adopting households of Bt cotton in Jalgaon, Maharashtra State, India, spanning the seasons 2002 and 2003. Results suggest that Bt adopters are indeed different from non-adopters in a number of ways. Adopters appear to specialize more on cotton (at least in terms of the land area they devote to the crop), spend more money on irrigation and grow well-performing non-Bt varieties of cotton (Bunny). Taking gross margin as the basis for comparison, Bt plots had 2.5 times the gross margin of non-Bt plots in both seasons. If only adopters are considered then the gross margin advantage of Bt plots reduces to 1.6 times that of non-Bt plots. This is still a significant advantage and could well explain the popularity of Bt in Maharashtra. However, it is clear that great care needs to be taken with such comparative studies.