Purification of recombinant proteins by chemical removal of the affinity tag.

The efficient removal of a N- or C-terminal purification tag from a fusion protein is necessary to obtain a protein in a pure and active form, ready for use in human or animal medicine. Current techniques based on enzymatic cleavage are expensive and result in the presence of additional amino acids at either end of the proteins, as well as contaminating proteases in the preparation. Here we evaluate an alternative method to the one-step affinity/protease purification process for large-scale purification. It is based upon the cyanogen bromide (CNBr) cleavage at a single methionine placed in between a histidine tag and a Plasmodium falciparum antigen. The C-terminal segment of the circumsporozoite polypeptide was expressed as a fusion protein with a histidine tag in Escherichia coli purified by Ni-NAT agarose column chromatography and subsequently cleaved by CNBr to obtain a polypeptide without any extraneous amino acids derived from the cleavage site or from the affinity purification tag. Thus, a recombinant protein is produced without the need for further purification, demonstrating that CNBr cleavage is a precise, efficient, and low-cost alternative to enzymatic digestion, and can be applied to large-scale preparations of recombinant proteins.

The Lincoln Highway Association’s “Object Lesson:” The Seedling Mile in Linn County Iowa,

The Seedling Mile in Linn County, Iowa, was part of the Lincoln Highway Association’s so-called “object lesson” program that sought to graphically demonstrate, in the paving of selected one-mile demonstration sections, the benefits of concrete paving to improving road travel across the nation. Constructed in 1918-19, this Seedling Mile became much more than an object lesson and served as something of a battleground between two municipalities—Marion and Cedar Rapids—in their struggle over the county seat and their place on the Lincoln Highway. The Seedling Mile eventually became part of a continuously paved section of the Lincoln Highway between Chicago and Cedar Rapids, with the whole of the Lincoln Highway in Iowa paved in some fashion by the 1930s. In 2002, Linn County reconstructed Mt. Vernon Road from the City of Mt. Vernon to the west end of the Seedling Mile impacting the historic road section. An agreement between concerned government agencies resulted in this publication in partial mitigation of the impact to this historic road section under the guidelines of the National Historic Preservation Act.

Violence as a research object and intervention in the health field: an analysis from the production of the Research Group on Gender, Health and Nursingi

The study aimed to describe how violence is revealed in the production of the Research Group on Gender, Health and Nursing. This is a historical research of qualitative approach, which evaluated the production of the Research Group, through content analysis. The results show gender as a central category in determining violence and health practices. This aspect determines limitations on professional practices of coping, such as the invisibility of the problem. The female autonomy, the use of alcohol and drugs and social vulnerability play an important relation with the phenomenon and the bond is revealed as potentiality of health practices to address the problem. Conclusion: The gender perspective in nursing research is an innovative field and counter-hegemonic, a possibility to assume a meaning of praxis by transforming potential of understanding and modes of intervention in the phenomenon of gender violence.

Sex differences: From mental rotation to object location memory, an ERP study

Sex differences in cognition have been largely investigated. The most consistent sex differences favoring females are observed in object location memory involving the left hemisphere whereas the most consistent sex differences favoring males are observed in tasks that require mental rotation involving the right hemisphere. Here we used a task involving these two abilities to see the impact of mental rotation on object location memory. To that end we used a combination of behavioral and event-related potential (ERP) electroencephalography (EEG) measures.A computer screen displayed a square frame of 4 pairs of images (a "teddy" bear, a shoe, an umbrella and a lamp) randomly arranged around a central fixation cross. After a 10-second interval for memorization, images disappeared and were replaced by a test frame with no image but a random pair of two locations marked in black. In addition, this test frame was randomly displayed either in the original orientation (0° rotation) or in the rotated one (90° clockwise - CW - or 90° counterclockwise - CCW). Preceding the test frame, an arrow indicating the presence or the absence of rotation of the frame was displayed on the screen. The task of the participants (15 females and 15 males) was to determine if two marked locations corresponded or not to a pair of identical images. Each response was followed by feedback.Findings showed no significant sex differences in the performance of the original orientation. In comparison with this position, the rotation of the frame produced an equal decrease of male and female performance. In addition, this decrease was significantly higher when the rotation of the frame was in a CCW direction. We further assessed the ERP when the arrow indicated the direction of rotation as stimulus-onset, during four time windows representing major components C1, P1, N1 and N2. Although no sex differences were observed in performance, brain activities differed according to sex. Enhanced amplitudes were found for the CCW compared to CW rotation over the right posterior areas for the P1, N1 and N2 components for men as well as for women. Major topographical differences related to sex were measured for the CW rotation condition as marked lateralized amplitude: left-hemisphere amplitude larger than right one was measured during P1 time range for men. These similar patterns prolonged from P1 to N1 for women. Early distinctions were found in interaction with sex between CCW and CW waveform amplitudes, expressing over anterior electrode sites during C1 time range (0-50 ms post-stimulus).In conclusion (i) women do not outperform men in object location memory in this study (absence of rotation condition); (ii) mental rotation, in particular the direction of rotation, influences performance on object location memory; (iii) CCW rotation is associated with activity in the right parietal hemisphere whereas the CW rotation involves the left parietal hemisphere; (iv) this last effect is less pronounced in males, which could explain why greater involvement of right parietal areas in men and of bilateral posterior areas in women is generally reported in mental rotation tasks; and (v) the early distinctions between both directions of rotation located over anterior sites could be related to sex differences in their respective involvement of control mechanisms.

Implementing interactive computing in an object-oriented environment

Statistical computing when input/output is driven by a Graphical User Interface is considered. A proposal is made for automatic control ofcomputational flow to ensure that only strictly required computationsare actually carried on. The computational flow is modeled by a directed graph for implementation in any object-oriented programming language with symbolic manipulation capabilities. A complete implementation example is presented to compute and display frequency based piecewise linear density estimators such as histograms or frequency polygons.

Development and use of a high-fidelity simulator for fetal endotracheal balloon occlusion (FETO) insertion and removal.

Objectives The objective of this article is to describe the development of an anatomically accurate simulator in order to aid the training of a perinatal team in the insertion and removal of a fetal endoscopic tracheal occlusion (FETO) balloon in the management of prenatally diagnosed congenital diaphragmatic hernia. Methods An experienced perinatal team collaborated with a medical sculptor to design a fetal model for the FETO procedure. Measurements derived from 28-week fetal magnetic resonance imaging were used in the development of an anatomically precise simulated airway within a silicone rubber preterm fetal model. Clinician feedback was then used to guide multiple iterations of the model with serial improvements in the anatomic accuracy of the simulator airway. Results An appropriately sized preterm fetal mannequin with a high-fidelity airway was developed. The team used this model to develop surgical skills with balloon insertion, and removal, and to prepare the team for an integrated response to unanticipated delivery with the FETO balloon still in situ. Conclusions This fetal mannequin aided in the ability of a fetal therapy unit to offer the FETO procedure at their center for the first time. This model may be of benefit to other perinatal centers planning to offer this procedure.

Effects of the removal of shelterwood on the foliar nutrient conceentrations of Norway spruce (Picea abies(L.)Karst.) on drained peatlands

Tiivistelmä: Neulasten pääravinnepitoisuuksien muutokset turvekankaan alikasvoskuusikossa ylispuuhakkuun jälkeen

Effects of convection on the removal of the multiplicity of stable states in one-dimensional vertical models

Here I develop a model of a radiative-convective atmosphere with both radiative and convective schemes highly simplified. The atmospheric absorption of radiation at selective wavelengths makes use of constant mass absorption coefficients in finite width spectral bands. The convective regime is introduced by using a prescribed lapse rate in the troposphere. The main novelty of the radiative-convective model developed here is that it is solved without using any angular approximation for the radiation field. The solution obtained in the purely radiation mode (i. e. with convection ignored) leads to multiple equilibria of stable states, being very similar to some results recently found in simple models of planetary atmospheres. However, the introduction of convective processes removes the multiple equilibria of stable states. This shows the importance of taking convective processes into account even for qualitative analyses of planetary atmosphere

Identification and removal of low-complexity sites in allele-specific analysis of ChIP-seq data.

MOTIVATION: High-throughput sequencing technologies enable the genome-wide analysis of the impact of genetic variation on molecular phenotypes at unprecedented resolution. However, although powerful, these technologies can also introduce unexpected artifacts. Results: We investigated the impact of library amplification bias on the identification of allele-specific (AS) molecular events from high-throughput sequencing data derived from chromatin immunoprecipitation assays (ChIP-seq). Putative AS DNA binding activity for RNA polymerase II was determined using ChIP-seq data derived from lymphoblastoid cell lines of two parent-daughter trios. We found that, at high-sequencing depth, many significant AS binding sites suffered from an amplification bias, as evidenced by a larger number of clonal reads representing one of the two alleles. To alleviate this bias, we devised an amplification bias detection strategy, which filters out sites with low read complexity and sites featuring a significant excess of clonal reads. This method will be useful for AS analyses involving ChIP-seq and other functional sequencing assays.

Where sound position influences sound object representations: a 7-T fMRI study.

Evidence from human and non-human primate studies supports a dual-pathway model of audition, with partially segregated cortical networks for sound recognition and sound localisation, referred to as the What and Where processing streams. In normal subjects, these two networks overlap partially on the supra-temporal plane, suggesting that some early-stage auditory areas are involved in processing of either auditory feature alone or of both. Using high-resolution 7-T fMRI we have investigated the influence of positional information on sound object representations by comparing activation patterns to environmental sounds lateralised to the right or left ear. While unilaterally presented sounds induced bilateral activation, small clusters in specific non-primary auditory areas were significantly more activated by contra-laterally presented stimuli. Comparison of these data with histologically identified non-primary auditory areas suggests that the coding of sound objects within early-stage auditory areas lateral and posterior to primary auditory cortex AI is modulated by the position of the sound, while that within anterior areas is not.

Effluent particle removal by microirrigation system filters

The aim of this work was to determine whether the filters used in microirrigation systems can remove potentially emitter-clogging particles. The particle size and volume distributions of different effluents and their filtrates were established, and the efficiency of the removal of these particles and total suspended solids by screen, disc and sand filters determined. In most of the effluents and filtrates, the number of particles with a diameter > 20 μm was minimal. By analysing the particle volume distribution it was found that particles larger than the disc and screen filter pores appeared in the filtrates. However, the sand filter was able to retain particles larger than the pore size. The filtration efficiency depended more on the type of effluent than on the filter. It was also found that the particle size distribution followed a potential law. Analysis of the β exponents showed that the filters did not significantly modify the particle size distribution of the effluents

Analysis of warm season thunderstorms using an object-oriented tracking method based on radar and total lightning data

Monitoring thunderstorms activity is an essential part of operational weather surveillance given their potential hazards, including lightning, hail, heavy rainfall, strong winds or even tornadoes. This study has two main objectives: firstly, the description of a methodology, based on radar and total lightning data to characterise thunderstorms in real-time; secondly, the application of this methodology to 66 thunderstorms that affected Catalonia (NE Spain) in the summer of 2006. An object-oriented tracking procedure is employed, where different observation data types generate four different types of objects (radar 1-km CAPPI reflectivity composites, radar reflectivity volumetric data, cloud-to-ground lightning data and intra-cloud lightning data). In the framework proposed, these objects are the building blocks of a higher level object, the thunderstorm. The methodology is demonstrated with a dataset of thunderstorms whose main characteristics, along the complete life cycle of the convective structures (development, maturity and dissipation), are described statistically. The development and dissipation stages present similar durations in most cases examined. On the contrary, the duration of the maturity phase is much more variable and related to the thunderstorm intensity, defined here in terms of lightning flash rate. Most of the activity of IC and CG flashes is registered in the maturity stage. In the development stage little CG flashes are observed (2% to 5%), while for the dissipation phase is possible to observe a few more CG flashes (10% to 15%). Additionally, a selection of thunderstorms is used to examine general life cycle patterns, obtained from the analysis of normalized (with respect to thunderstorm total duration and maximum value of variables considered) thunderstorm parameters. Among other findings, the study indicates that the normalized duration of the three stages of thunderstorm life cycle is similar in most thunderstorms, with the longest duration corresponding to the maturity stage (approximately 80% of the total time).