bluuebunny/crossref_metadata_2025 huggingface dataset:
CRIg signals induce anti‐intracellular bacterial phagosome activity in a chloride intracellular channel 3‐dependent manner
Authors: Kim Kwang H. , Choi Beom K. , Song Keoung M. , Cha Ki W. , Kim Young H. , Lee Ho , Han In‐Seob , Kwon Byoung S.
Published: December 2012

Macrophages provide a first line of defense against bacterial infection by engulfing and killing invading bacteria, but intracellular bacteria such as Listeria monocytogenes (LM) can survive in macrophages by various mechanisms of evasion. Complement receptor of the immunoglobulin (CRIg), a C3b receptor, binds to C3b on opsonized bacteria and facilitates clearance of the bacteria by promoting their uptake. We found that CRIg signaling induced by agonistic anti‐CRIg mAb enhanced the killing of intracellular LM by macrophages, and that this occurred in LM‐containing phagosomes. Chloride intra‐cellular channel 3 CLIC3, an intracellular chloride channel protein, was essential for CRIg‐mediated LM killing by directly interacting with the cytoplasmic domain of CRIg, and the two proteins colocalized on the membranes of LM‐containing vacuoles. CLIC3−/− mice were as susceptible to LM as CRIg−/− mice. These findings identify a mechanism embedded in the process by which macrophages take up opsonized bacteria that prevents the bacteria from evading cell‐mediated killing.
Cytomegalovirus‐specific T cells are detectable in early childhood and allow assignment of the infection status in children with passive maternal antibodies
Authors: Ritter Marion , Schmidt Tina , Dirks Jan , Hennes Pia , Juhasz‐Böss Ingolf , Solomayer Erich F. , Gortner Ludwig , Gärtner BarbaraC. , Rohrer Tilman , Sester Urban , Sester Martina
Published: December 2012

Serological identification of the cytomegalovirus (CMV) status in children less than 18 months of age is complicated by the variable persistence of maternal antibodies. As T cells are not passively transferred, we attempted to assess whether CMV‐specific cellular immunity may be superior to determine the actual CMV status; we also performed a functional characterization of T‐cell immunity in childhood. Antibodies from 59 mothers and 168 children were determined, and specific CD4+ T cells were identified by induction of IFN‐γ, IL‐2, TNF‐α, IL‐4, and IL‐17 after CMV‐specific and polyclonal stimulation. Agreement between both tests was perfect for mothers and children more than 18 months. Among infants less than 18 months, 17/30 were concordantly negative. Interestingly, 8/13 seropositive children had detectable CMV‐specific T cells, whereas only 5/13 were T‐cell negative, indicating passive immunity. CMV‐specific T cells from young infants differed in cytokine profiles from that of older age groups, and polyclonal effector T‐cell frequencies were higher in young infants with detectable CMV‐specific T cells compared with those without. In conclusion, the majority of young infants with CMV‐specific antibodies show evidence of true infection, which indicates that passive immunity is overestimated. Our data may have important implications for improved risk stratification and CMV management in infants in the setting of transplantation.
Gel electrophoresis of DNA partially denatured at the ends: What are the dominant conformations?
Authors: Sean David , Slater Gary W.
Published: December 2012

Gel electrophoresis of a partially denatured dsDNA fragment is studied using Langevin Dynamics computer simulations. For simplicity, the denatured ssDNA sections are placed at the ends of the fragment in a symmetrical fashion. A squid‐like conformation is found to sometimes cause the fragment to completely block in the gel. In fact, this conformation is the principal cause of the steep reduction in mobility observed in the simulations. As the field is increased, it is found that the occurrence of this conformation dominates the migration dynamics. Although the squid conformation seems to be more stable at high fields, the field can eventually force the fragments to thread through the gel pores regardless. We qualitatively explore the behavior of this squid‐like conformation across a range of fields and degrees of denaturation, and we discuss the relevance of our findings for TGGE.
Ecological Risk‐O‐Meter: a risk assessor and manager software tool for better decision making in ecosystems
Authors: Sahinoglu Mehmet , Simmons Susan J. , Cahoon Lawrence B. , Morton Scott
Published: December 2012

Increased awareness of environmental issues and their effects on ecological systems and human health drive an interest in developing computational methods to reduce detrimental consequences. For example, there are concerns regarding chlorofluorocarbons and their impact on stratospheric ozone, radon and its effect on human health, coal mining and effects on habitat loss, as well as numerous other issues. However, these issues do not exist in a vacuum nor occur just one at a time. There is a need to assess social and ecological risks comprehensively and account for numerous, inter‐related potential risks. Given limited funds available for addressing these issues, how can spending for purposes of environmental and ecological mitigation be optimized? What is the magnitude of overall ecological risk for a given region? Novel software, the “Ecological Risk‐o‐Meter”, addresses these questions and concerns. The software tool not only assesses the current environmental and ecological risks, but also takes into account potential solutions and provides guidance as to how spending can be optimized to reducing overall environmental risk. We demonstrate this new tool and show how to optimize the costs of risk reduction in recursive cycles based on feedbacks. Copyright © 2012 John Wiley & Sons, Ltd.
Photocatalytic Decontamination of Sulfur Mustard Over Manganese Oxide Nanobelts
Authors: Ramacharyulu P.V.R.K. , Prasad G.K. , Kumar J. Praveen , Ganesan K. , Singh Beer , Dwivedi K.
Published: December 2012

Photocatalytic decontamination of sulfur mustard was studied on manganese oxide nanobelts (MON) in the presence of Sun light and UV‐A light. Data obtained from the experiments conducted under the irradiation of Sun light and UV‐A light was compared with that of the obtained without irradiation of light. Sulfur mustard and the products of reaction were studied using gas chromatography, gas chromatography‐mass spectrometry techniques (GC‐MS). 100% of sulfur mustard was found to be decontaminated on the surface of MON with irradiation of Sun light in 8 h. Whereas without irradiation only 46% of sulfur mustard was found to be decontaminated. Superior Sun light activity of MON against sulfur mustard was attributed to its high irradiance. GC‐MS data indicated the formation of acetaldehyde, carbon dioxide, sulfur mustard sulfoxide, chloroethyl vinyl sulfide, and so forth, due to photocatalysis of sulfur mustard. © 2012 American Institute of Chemical Engineers Environ Prog, 32: 1118–1123, 2013
Optimization of Biosorption Conditions for Color Removal by Taguchi DOE Methodology
Authors: Deniz Fatih
Published: December 2012

The biosorption conditions for the removal of Maxilon Red GRL (MR GRL) dye by walnut shell were optimized using the Taguchi technique. This study adopted an L9 orthogonal array based on the Taguchi method to conduct a series of experiments and statistically evaluated the experimental data by the analysis of variance (ANOVA). The main operating parameters including biosorbent dosage, initial dye concentration, and contact time were chosen to achieve the maximum dye removal. The best conditions were found as initial dye concentration of 80 mg L−1, contact time of 120 min and biosorbent dose of 1 g L−1 for the larger is better response category. The biosorbent dosage was found to be the most important factor on the dye removal. The optimal conditions predicted were further validated by the confirmation experiments. In this way, the Taguchi approach proved to bea reliable tool in the optimizing biosorption conditions. ©2012 American Institute of Chemical Engineers Environ Prog, 32: 1129–1133, 2013
Adsorption kinetics, mechanism, isotherm, and thermodynamic analysis of copper ions onto the surface modified agricultural waste
Authors: Senthil Kumar P. , Senthamarai C. , Durgadevi A.
Published: December 2012

The adsorption of copper ions onto the surface modified Strychnos potatorum seeds (SMSP) in a batch adsorption system has been studied. Kinetics, mechanism, isotherm, and thermodynamic parameters have been estimated. Adsorption kinetics of copper ions onto the SMSP follows a pseudo‐second order kinetic model. Adsorption mechanism was explained with the intraparticle diffusion model, Boyd kinetic model (BKM), and Shrinking core model (SCM). Adsorption process was found to be controlled by both intraparticle diffusion and film diffusion. The diffusivity values were estimated from the BKM and SCM. Adsorption isotherm data agreed well with the Freundlich adsorption isotherm model, which indicates the multilayer adsorption of copper ions onto the SMSP. The maximum monolayer adsorption capacity of the SMSP was found to be 248 mg of copper ions/g of SMSP. Adsorption thermodynamic studies show that the adsorption of copper ions onto the SMSP was spontaneous and exothermic in nature. © 2012 American Institute of Chemical Engineers Environ Prog, 33: 28–37, 2014
The application of COSI‐Corr to determine dune system dynamics in the southern Namib Desert using ASTER data
Authors: Scheidt Stephen P. , Lancaster Nick
Published: December 2012

The Coregistration of Optically Sensed Images and Correlation (COSI‐Corr) algorithm was used to estimate dune migration and sand flux rates from a series of remotely‐sensed optical image pairs. Several areas of barchan and transverse dunes transport sand along definite pathways, feeding the southern part of the Namib Sand Sea from beaches and deflationary basins in the Sperrgebiet. We give a detailed description of the pre‐ and post‐processing routines used for our COSI‐Corr analysis, and we evaluate the sub‐pixel dune migration results at the dune and dune‐field scales. The best set of parameters for the application of the algorithm was systematically derived, resulting in methodological refinements of the cross‐correlation windows. The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) sensor aboard the Terra satellite provided the remote sensing imagery. Few comparisons have been made between dune migration measurements using COSI‐Corr and other types of change detection analysis, and few dune fields have served as test cases for COSI‐Corr. We suggest that analysis and interpretation of COSI‐Corr output from ASTER data is more valuable if combined with other image analysis techniques, such as manual digitization of dunes in a geographic information system (GIS), processing of write memory function insertion (WMFI) imagery and multispectral image (MSI) analysis of composition. The dune migrations estimated from COSI‐Corr results were validated using GIS and WMFI, and later MSI analysis added an important regional, contextual framework of sand transport pathways. The synthesis of these works lends more confidence to understanding the Namib dune system's dynamics and provides a basis for future comparisons to other dune fields. Copyright © 2013 John Wiley & Sons, Ltd.
Towards petascale spectral simulations for transition analysis in wall bounded flow
Authors: Montagnier Julien , Cadiou Anne , Buffat Marc , Penven Lionel Le
Published: December 2012

An efficient parallel spectral method for direct numerical simulations of transitional and turbulent flows is described in this paper. The parallelization is classically based on a bidimensional domain decomposition, but has been specifically developed for a solenoidal Fourier–Chebyshev spectral approximation where in one Fourier direction, the number of modes is very large compared with the two other directions. The approach therefore differs from classical libraries developed for cubic Fourier boxes. The strategy uses message‐passing interface (MPI) for message‐passing among nodes and is fairly portable. One of the originalities of this paper is the use of an efficient hybrid programming with MPI for internodes communications and a coarse grain parallelism using OpenMP for core shared‐memory computation, instead of the classical hybrid programming with MPI and a fine granularity parallelism at the loop level with OpenMP directives. This hybrid parallelism has been tested on the recent generation of high‐performance parallel supercomputers involving a few tens of cores per node. Performances are evaluated on different low‐frequency and high‐frequency processors massively parallel platforms. We demonstrate that spectral methods, which are known to be inherently ill‐fitted for the new generation of high‐performance distributed‐memory computers, can be implemented efficiently using this hybrid programming with good scalability and a very fast wall‐clock time per iteration. New numerical experiments are therefore now accessible on petascale computers, while keeping the attractive features of spectral methods such as accuracy, exponential convergence, computational efficiency and conservative properties. This is illustrated by a direct numerical simulation of the transition of the boundary layers developing from the entrance section of a plane channel and interacting to merge into a fully turbulent flow. Copyright © 2012 John Wiley & Sons, Ltd.
Implementation of slip boundary conditions in the finite volume method: new techniques
Authors: Ferrás L.L. , Nóbrega J.M. , Pinho F.T.
Published: December 2012

Two different techniques for the implementation of the linear and nonlinear slip boundary conditions into a finite volume method based numerical code are presented. For the linear Navier slip boundary condition, an implicit implementation in the system of equations is carried out for which there is no need for any relaxation, especially when handling high slip coefficients. For three different nonlinear slip boundary conditions, two different methods are devised, one based on solving a transcendental equation for the boundary and the other on the linearization of the slip law. For assessment purposes, comparison is made between these new methods and the usual iterative process. With these new methods, the convergence difficulties, typical of the iterative procedure, are eliminated, and for some of the test cases, the convergence rate even increased with the slip velocity. The details of these implementations are given first for a simple geometry using orthogonal meshes and Cartesian coordinates followed by their generalization to non‐Cartesian coordinates and nonorthogonal meshes. The developed code was tested in the benchmark slip‐stick and 4:1 contraction flows, evidencing the robustness of the proposed procedures. Copyright © 2013 John Wiley & Sons, Ltd.
Sequence Kernel Association Test for Quantitative Traits in Family Samples
Authors: Chen Han , Meigs James B. , Dupuis Josée
Published: December 2012

A large number of rare genetic variants have been discovered with the development in sequencing technology and the lowering of sequencing costs. Rare variant analysis may help identify novel genes associated with diseases and quantitative traits, adding to our knowledge of explaining heritability of these phenotypes. Many statistical methods for rare variant analysis have been developed in recent years, but some of them require the strong assumption that all rare variants in the analysis share the same direction of effect, and others requiring permutation to calculate the P‐values are computer intensive. Among these methods, the sequence kernel association test (SKAT) is a powerful method under many different scenarios. It does not require any assumption on the directionality of effects, and statistical significance is computed analytically. In this paper, we extend SKAT to be applicable to family data. The family‐based SKAT (famSKAT) has a different test statistic and null distribution compared to SKAT, but is equivalent to SKAT when there is no familial correlation. Our simulation studies show that SKAT has inflated type I error if familial correlation is inappropriately ignored, but has appropriate type I error if applied to a single individual per family to obtain an unrelated subset. In contrast, famSKAT has the correct type I error when analyzing correlated observations, and it has higher power than competing methods in many different scenarios. We illustrate our approach to analyze the association of rare genetic variants using glycemic traits from the Framingham Heart Study.
SNP Prioritization Using a Bayesian Probability of Association
Authors: Thompson John R. , Gögele Martin , Weichenberger Christian X. , Modenese Mirko , Attia John , Barrett Jennifer H. , Boehnke Michael , De Grandi Alessandro , Domingues Francisco S. , Hicks Andrew A. , Marroni Fabio , Pattaro Cristian , Ruggeri Fabrizio , Borsani Giuseppe , Casari Giorgio , Parmigiani Giovanni , Pastore Andrea , Pfeufer Arne , Schwienbacher Christine , Taliun Daniel , Consortium CKDGen , Fox Caroline S. , Pramstaller Peter P. , Minelli Cosetta
Published: December 2012

Prioritization is the process whereby a set of possible candidate genes or SNPs is ranked so that the most promising can be taken forward into further studies. In a genome‐wide association study, prioritization is usually based on the P‐values alone, but researchers sometimes take account of external annotation information about the SNPs such as whether the SNP lies close to a good candidate gene. Using external information in this way is inherently subjective and is often not formalized, making the analysis difficult to reproduce. Building on previous work that has identified 14 important types of external information, we present an approximate Bayesian analysis that produces an estimate of the probability of association. The calculation combines four sources of information: the genome‐wide data, SNP information derived from bioinformatics databases, empirical SNP weights, and the researchers’ subjective prior opinions. The calculation is fast enough that it can be applied to millions of SNPS and although it does rely on subjective judgments, those judgments are made explicit so that the final SNP selection can be reproduced. We show that the resulting probability of association is intuitively more appealing than the P‐value because it is easier to interpret and it makes allowance for the power of the study. We illustrate the use of the probability of association for SNP prioritization by applying it to a meta‐analysis of kidney function genome‐wide association studies and demonstrate that SNP selection performs better using the probability of association compared with P‐values alone.
How ‘love’ and ‘hate’ differ from ‘sleep’: Using combined electro/magnetoencephalographic data to reveal the sources of early cortical responses to emotional words
Authors: Keuper Kati , Zwanzger Peter , Nordt Marisa , Eden Annuschka , Laeger Inga , Zwitserlood Pienie , Kissler Johanna , Junghöfer Markus , Dobel Christian
Published: December 2012

Emotional words—as symbols for biologically relevant concepts—are preferentially processed in brain regions including the visual cortex, frontal and parietal regions, and a corticolimbic circuit including the amygdala. Some of the brain structures found in functional magnetic resonance imaging are not readily apparent in electro‐ and magnetoencephalographic (EEG; MEG) measures. By means of a combined EEG/MEG source localization procedure to fully exploit the available information, we sought to reduce these discrepancies and gain a better understanding of spatiotemporal brain dynamics underlying emotional‐word processing. Eighteen participants read high‐arousing positive and negative, and low‐arousing neutral nouns, while EEG and MEG were recorded simultaneously. Combined current‐density reconstructions (L2‐minimum norm least squares) for two early emotion‐sensitive time intervals, the P1 (80–120 ms) and the early posterior negativity (EPN, 200–300 ms), were computed using realistic individual head models with a cortical constraint. The P1 time window uncovered an emotion effect peaking in the left middle temporal gyrus. In the EPN time window, processing of emotional words was associated with enhanced activity encompassing parietal and occipital areas, and posterior limbic structures. We suggest that lexical access, being underway within 100 ms, is speeded and/or favored for emotional words, possibly on the basis of an “emotional tagging” of the word form during acquisition. This gives rise to their differential processing in the EPN time window. The EPN, as an index of natural selective attention, appears to reflect an elaborate interplay of distributed structures, related to cognitive functions, such as memory, attention, and evaluation of emotional stimuli. Hum Brain Mapp 35:875–888, 2014. © 2012 Wiley Periodicals, Inc.
Spatial heterogeneity of the relation between resting‐state connectivity and blood flow: An important consideration for pharmacological studies
Authors: Khalili‐Mahani Najmeh , van Osch Matthias J. , de Rooij Mark , Beckmann Christian F. , van Buchem Mark A. , Dahan Albert , van Gerven Johannes M. , Rombouts Serge A.R.B.
Published: December 2012

Resting state fMRI (RSfMRI) and arterial spin labeling (ASL) provide the field of pharmacological Neuroimaging tool for investigating states of brain activity in terms of functional connectivity or cerebral blood flow (CBF). Functional connectivity reflects the degree of synchrony or correlation of spontaneous fluctuations—mostly in the blood oxygen level dependent (BOLD) signal—across brain networks; but CBF reflects mean delivery of arterial blood to the brain tissue over time. The BOLD and CBF signals are linked to common neurovascular and hemodynamic mechanisms that necessitate increased oxygen transportation to the site of neuronal activation; however, the scale and the sources of variation in static CBF and spatiotemporal BOLD correlations are likely different. We tested this hypothesis by examining the relation between CBF and resting‐state‐network consistency (RSNC)—representing average intranetwork connectivity, determined from dual regression analysis with eight standard networks of interest (NOIs)—in a crossover placebo‐controlled study of morphine and alcohol. Overall, we observed spatially heterogeneous relations between RSNC and CBF, and between the experimental factors (drug‐by‐time, time, drug and physiological rates) and each of these metrics. The drug‐by‐time effects on CBF were significant in all networks, but significant RSNC changes were limited to the sensorimotor, the executive/salience and the working memory networks. The post‐hoc voxel‐wise statistics revealed similar dissociations, perhaps suggesting differential sensitivity of RSNC and CBF to neuronal and vascular endpoints of drug actions. The spatial heterogeneity of RSNC/CBF relations encourages further investigation into the role of neuroreceptor distribution and cerebrovascular anatomy in predicting spontaneous fluctuations under drugs. Hum Brain Mapp 35:929–942, 2014. © 2012 Wiley Periodicals, Inc.
GENERAL BUDGET SUPPORT: HAS IT BENEFITED THE HEALTH SECTOR?
Authors: Fernandes Antunes Adelio , Xu Ke , James Chris D. , Saksena Priyanka , Van de Maele Nathalie , Carrin Guy , Evans David B.
Published: December 2012

There has been recent controversy about whether aid directed specifically to health has caused recipient governments to reallocate their own funds to non‐health areas. At the same time, general budget support (GBS) has been increasing. GBS allows governments to set their own priorities, but little is known about how these additional resources are subsequently used. This paper uses cross‐country panel data to assess the impact of GBS programmes on health spending in low‐income and middle‐income countries, using dynamic panel techniques to estimate unbiased coefficients in the presence of serial correlation. We found no clear evidence that GBS had any impact, positive or negative, on government health spending derived from domestic sources. GBS also had no observed impact on total government health spending from all sources (external as well as domestic). In contrast, health‐specific aid was associated with a decline in health expenditures from domestic sources, but there was not a full substitution effect. That is, despite this observed fungibility, health‐specific aid still increases total government health spending from all sources. Finally, increases in total government expenditure led to substantial increases in domestic government health expenditures. Copyright © 2012 John Wiley & Sons, Ltd.
Recent climatic trends and linkages to river discharge in Central Vietnam
Authors: Souvignet Maxime , Laux Patrick , Freer Jim , Cloke Hannah , Thinh Dang Quang , Thuc Tran , Cullmann Johannes , Nauditt Alexandra , Flügel Wolfgang‐Albert , Kunstmann Harald , Ribbe Lars
Published: December 2012

In this study, change in rainfall, temperature and river discharge are analysed over the last three decades in Central Vietnam. Trends and rainfall indices are evaluated using non‐parametric tests at different temporal levels. To overcome the sparse locally available network, the high resolution APHRODITE gridded dataset is used in addition to the existing rain gauges. Finally, existing linkages between discharge changes and trends in rainfall and temperature are explored.

Results are indicative of an intensification of rainfall (+15%/decade), with more extreme and longer events. A significant increase in winter rainfall and a decrease in consecutive dry days provides strong evidence for a lengthening wet season in Central Vietnam. In addition, trends based on APHRODITE suggest a strong orographic signal in winter and annual trends. These results underline the local variability in the impacts of climatic change at the global scale. Consequently, it is important that change detection investigations are conducted at the local scale. A very weak signal is detected in the trend of minimum temperature (+0.2°C/decade). River discharge trends show an increase in mean discharge (31 to 35%/decade) over the last decades. Between 54 and 74% of this increase is explained by the increase in precipitation. The maximum discharge also responds significantly to precipitation changes leading to a lengthened wet season and an increase in extreme rainfall events. Such trends can be linked with a likely increase in floods in Central Vietnam, which is important for future adaptation planning and management and flood preparedness in the region. Copyright © 2012 John Wiley & Sons, Ltd.
Towards hydrological model calibration and validation: simulation of stable water isotopes using the isoWATFLOOD model
Authors: Stadnyk T. A. , Delavau C. , Kouwen N. , Edwards T. W. D.
Published: December 2012

Calibration and validation of hydrological models is a challenge, particularly in remote regions that are minimally gauged. This paper develops a novel methodology for large‐scale (>1000 km2) hydrological model calibration and validation using stable water isotopes founded on the rigorous constraints imposed by the need to conserve both water mass and stable isotopes simultaneously. The isoWATFLOOD model is applied to five basins within the Fort Simpson, Northwest Territories region of northern Canada to simulate stream discharge and oxygen‐18 signals over a 3‐year period. The isotopic variation of river discharge, runoff components, and evaporative fractionation are successfully simulated on both a seasonal and continual basis over the watershed domain to demonstrate the application of isotope tracers to regional hydrologic calibration. The intended application of this research is to remote, large‐scale basins, showing promise for improving predictions in minimally gauged basins and climate change research where traditional, rigorous approaches to constraining parameter uncertainty may be impractical. This coupled isotope‐hydrological (i.e. iso‐hydrological) approach to modelling reduces the number of possible parameterizations, resulting in potentially more physically‐based hydrological predictions. isoWATFLOOD provides a tool for water resource managers and utilities to use operationally for water use, allocation, and runoff generation estimations. Copyright © 2012 John Wiley & Sons, Ltd.
Effect of CoCl2 on the content of different metals and a relative activity of DNA‐hydrolyzing abzymes in the blood plasma of mice
Authors: Legostaeva Galina A. , Zaksas Nataliya P. , Gluhcheva Yordanka G. , Sedykh Sergey E. , Madzharova Maria E. , Atanassova Nina N. , Buneva Valentina N. , Nevinsky Georgy A.
Published: December 2012

Cobalt is a transition metal and an essential trace element that is required for vitamin B12 biosynthesis, enzyme activation, and so on but is toxic in high concentrations. It was shown that the content of different elements in the plasma of 2‐month‐old BALB/c mice (control group) decreased in the following order: Ca > Mg > Si > Fe > Zn > Cu ≥ Al ≥ B. The treatment of mice with CoCl2 did not appreciably change the relative content of Ca, Cu, and Zn, but a significant increase in the content of B (2.3‐fold), Mg (1.5‐fold), Al and Fe (2.1‐fold), and Si (3.4‐fold) was found. The treatment of mice led to a 2.2‐fold decrease in the concentration of the total blood protein and a 1.7 ± 0.2‐fold decrease of total immunoglobulin Gs (IgGs). Deoxyribonuclease IgGs corresponding to mice treated (t‐IgGs) and non‐treated (nt‐IgGs) with CoCl2 contained intrinsically bound metal ions; these IgGs hydrolyzed DNA with very low activity but were not active in the presence of ethylenediaminetetraacetic acid or after Ab dialysis against ethylenediaminetetraacetic acid. The average RAs of deoxyribonuclease nt‐IgGs increased after addition of external metal ions in the following order: Zn2+ < Ca2+ < Cu2+ < Fe2+ < Mn2+ < Mg2+ < Co2+ < Ni2+. Interestingly, t‐IgGs demonstrated lower activities than those for nt‐IgGs either in the absence of external metal ions (2.7‐fold) or in the presence of Cu2+ (9.5‐fold) > Co2+ (5.6‐fold) > Zn2+ (5.1‐fold) > Mg2+ (4.1‐fold) > Ca2+ (3.0‐fold) > Fe2+ (1.3‐fold). However, the RAs of t‐IgGs were remarkably more active than nt‐IgGs in the presence of best activators of t‐IgGs Ni2+ (1.4‐fold) and especially Mn2+ (2.2‐fold). The data may be useful for an understanding of Co toxicity, its effect on the concentration of other metal ions, and a change of metal‐dependent specificity of Abzs. Copyright © 2012 John Wiley & Sons, Ltd.
Glutathione S‐transferase π complexes with and stimulates Na+,K+‐ATPase
Authors: Ochiai Hideo , Eguchi Hiroshi , Noguchi Shunsuke , Hayashi Yutaro , Nishino Hideaki , Kawamura Masaru , Wu Chau H.
Published: December 2012

Glutathione S‐transferase (GST) was found to complex with the Na+,K+‐ATPase as shown by binding assay using quartz crystal microbalance. The complexation was obstructed by the addition of antiserum to the α‐subunit of the Na+,K+‐ATPase, suggesting the specificity of complexation between GST and the Na+,K+‐ATPase. Co‐immunoprecipitation experiments, using the anti‐α‐subunit antiserum to precipitate the GST‐Na+,K+‐ATPase complex and then using antibodies specific to an isoform of GST to identify the co‐precipitated proteins, revealed that GSTπ was complexed with the Na+,K+‐ATPase. GST stimulated the Na+,K+‐ATPase activity up to 1.4‐fold. The level of stimulation exhibited a saturable dose–response relationship with the amount of GST added, although the level of stimulation varied depending on the content of GSTπ in the lots of GST received from supplier. The stimulation was also obtained when recombinant GSTπ was used, confirming the results. When GST was treated with reduced glutathione, GST activity was greatly stimulated, whereas the level of stimulation of the Na+,K+‐ATPase activity was similar to that when untreated GST was added. When GST was treated with H2O2, GST activity was greatly diminished while the stimulation of the Na+,K+‐ATPase activity was preserved. The results suggest that GSTπ complexes with the Na+,K+‐ATPase and stimulates the latter independent of its GST activity. Copyright © 2012 John Wiley & Sons, Ltd.
Characterization of selenium‐containing glutathione transferase zeta1–1 with high GPX activity prepared in eukaryotic cells
Authors: Yin Li , Song Jian , Board Philip G. , Yu Yang , Han Xiao , Wei Jingyan
Published: December 2012

Accumulating evidence shows that glutathione peroxidase (GPX, EC.1.11.1.9), one of the most important antioxidant selenoenzymes, plays an essential role in protecting cells and tissues against oxidative damage by catalyzing the reduction of hydrogen peroxide by glutathione. Unfortunately, because of the limited availability and poor stability of GPX, it has not been used clinically to protect against oxidative stress. To overcome these problems, it is necessary to generate mimics of GPX. In this study, we have used directed mutagenesis and the inclusion of a selenocysteine (Sec) insertion sequence to engineer the expression in eukaryotic cells of human glutathione transferase zeta1–1 (hGSTZ1–1) with Sec in the active site (seleno‐hGSTZ1–1). This modification converted hGSTZ1–1 into an active GPX and is the first time this has been achieved in eukaryotic cells. The GPX activity of seleno‐hGSTZ1–1 is higher than that of GPX from bovine liver, indicating Sec at the active site plays an important role in the determination of catalytic specificity and performance. Kinetic studies revealed that the ping–pong catalytic mechanism of Se‐hGSTZ1–1 is similar to that of the natural GPX. Copyright © 2012 John Wiley & Sons, Ltd.
Molecular recognition force spectroscopy study of the dynamic interaction between aptamer GBI‐10 and extracellular matrix protein tenascin‐C on human glioblastoma cell
Authors: Li Yongjun , Qiao Haiyan , Yan Wei , Zhang Jing , Xing Chunyan , Wang Hongda , Zhang Bailin , Tang Jilin
Published: December 2012

Molecular recognition force spectroscopy (MR‐FS) was applied to investigate the dynamic interaction between aptamer GBI‐10 and tenascin‐C (TN‐C) on human glioblastoma cell surface at single‐molecule level. The unbinding force between aptamer GBI‐10 and TN‐C was 39 pN at the loading rate of 0.3 nN sec−1. A series of kinetic parameters concerning interaction process such as the unbinding force fu, the association rate constant kon, dissociation rate constant at zero force koff, and dissociation constant KD for aptamer GBI‐10/TN‐C complexes were acquired. In addition, the interaction of aptamer GBI‐10 with TN‐C depended on the presence of Mg2+. This work demonstrates that MR‐FS can be used as an attractive tool for exploring the interaction forces and dynamic process of aptamer and ligand at the single‐molecule level. As a future perspective, MR‐FS may be used as a potential diagnostic and therapeutic tool by combining with other techniques. Copyright © 2012 John Wiley & Sons, Ltd.
Halilectin 1 (H‐1) and Halilectin 2 (H‐2): two new lectins isolated from the marine sponge Haliclona caerulea
Authors: Carneiro Rômulo Farias , de Melo Arthur Alves , Nascimento Fernando Edson Pessoa do , Simplicio Clareane Avelino , Nascimento Kyria Santiago do , Rocha Bruno Anderson Matias da , Saker‐Sampaio Silvana , Moura Raniere da Mata , Mota Sula Salani , Cavada Benildo Sousa , Nagano Celso Shiniti , Sampaio Alexandre Holanda
Published: December 2012

Two new lectins named Halilectin 1 (H‐1) and Halilectin 2 (H‐2) were isolated from the marine sponge Haliclona caerulea using a combination of affinity chromatography on stroma fixed onto Sephadex G‐25 and cation and anion exchange chromatography. H‐1 is a monomeric protein with a molecular mass of 40 kDa estimated using sodium dodecyl sulfate polyacrylamide gel electrophoresis and 15 kDa estimated using a TSK gel. Conversely, H‐2 is a homodimeric protein with 15 kDa monomers linked via weak interactions. H‐1 more effectively agglutinates trypsinized rabbit erythrocytes, whereas H‐2 more effectively agglutinates native rabbit erythrocytes. The hemagglutinating activity of H‐1 could be not inhibited by any tested sugars, but H‐2 was inhibited by orosomucoid and porcine stomach mucin. Neither lectin was dependent on divalent ions. H‐1 was stable at basic pH range and temperatures up to 50 °C, whereas H‐2 was stable at acid pH range and temperatures up to 80 °C. The H. caerulea lectins exhibited dose‐dependent toxicity against Artemia nauplii. Additionally, 76% of the primary structure of H‐2 was determined using tandem mass spectrometry to contain a unique amino acid sequence with no similarity to any members of the animal lectin family. Copyright © 2012 John Wiley & Sons, Ltd.
Comparison of oncogenic HPV type‐specific viral DNA load and E6/E7 mRNA detection in cervical samples: Results from a multicenter study
Authors: Broccolo Francesco , Fusetti Lisa , Rosini Sandra , Caraceni Donatella , Zappacosta Roberta , Ciccocioppo Lucia , Matteoli Barbara , Halfon Philippe , Malnati Mauro S. , Ceccherini‐Nelli Luca
Published: December 2012

High‐risk human papillomavirus (HR‐HPV) genotype viral load and E6/E7 mRNA detection are proposed as surrogate markers of malignant cervical lesion progression. Currently, the use of commercially available DNA‐based or mRNA‐based tests is under investigation. In this study, the viral DNA load and E6/E7 mRNA detection of the five most common HR‐HPV types detected in cervical cancer worldwide were compared in 308 cervical samples by using in‐house type‐specific quantitative real‐time PCR assays and PreTect HPV‐Proofer test, respectively. Sensitivity and negative predictive values were higher for the HPV‐DNA assays combined (95.0% and 96.0%, respectively) than the RNA assays (77.0% and 88.0%, respectively); conversely, the mRNA test showed a higher specificity and higher positive predictive value (81.7% and 66.9%, respectively) than the DNA test (58.6% and 52.5%, respectively) for detecting histology‐confirmed high‐grade cervical intraepithelial neoplasia. A significantly higher association between viral DNA load and severity of disease was observed for HPV 16 and 31 (γ = 0.62 and γ = 0.40, respectively) than for the other HPV types screened. A good degree of association between the two assays was found for detection of HPV 16 (k = 0.83), HPV 18 (k = 0.72), HPV 33 (k = 0.66), and HPV 45 (k = 0.60) but not for HPV 31 (k = 0.24). Sequence analysis in L1 and E6‐LCR regions of HPV 31 genotypes showed a high level of intra‐type variation. HR‐HPV viral DNA load was significantly higher in E6/E7 mRNA positive than negative samples (P < 0.001), except for HPV 31. These findings suggest that transcriptional and replicative activities can coexist within the same sample. J. Med. Virol. 85:472–482, 2013. © 2012 Wiley Periodicals, Inc.
Does high viral load of hepatitis E virus influence the severity and prognosis of acute liver failure during pregnancy?
Authors: Borkakoti Jayanta , Hazam Rajib Kishore , Mohammad Asim , Kumar Ashok , Kar Premashis
Published: December 2012

The incidence and mortality in pregnant women with acute liver failure caused by hepatitis E virus (HEV) is high. Data on the viral load of HEV during pregnancy are limited. The study was designed to determine the viral load of HEV and its association with the disease severity in patients with acute liver failure. A total of HEV related 163 patients with acute liver failure which included 105 pregnant, 46 non‐pregnant women and girls and 12 men and 730 patients with acute viral hepatitis which comprised of 220 pregnant women; 282 non‐pregnant women and girls and 228 men were included. Viral load was measured by real‐time PCR. Comparison was made between the pregnant and non‐pregnant women. HEV RNA was detectable in 265 patients (142 pregnant; 75 non‐pregnant and 48 men) and 104 patients with acute liver failure (64 pregnant, 34 non‐pregnant and 6 men). The viral load of HEV in pregnant women with acute liver failure and acute viral hepatitis was significantly higher 129,984.0 ± 103,104.17 and 768.92 ± 1,105.40 copies/ml, respectively compared to the non‐pregnant women which was 189.2 ± 225 and 12.73 ± 7.8 copies/ml (P < 0.0001). The viral load of HEV was also significantly higher in the pregnant patients with acute liver failure compared to the pregnant women with acute viral hepatitis and also men (P < 0.0001). High viral load of HEV during pregnancy could be one of the factors responsible for the severity of the infection during pregnancy. J. Med. Virol. 85:620–626, 2013. © 2012 Wiley Periodicals, Inc.
Dichlorvos‐induced cell cycle arrest and DNA damage repair activation in primary rat microglial cells
Authors: Sunkaria Aditya , Wani Willayat Yousuf , Sharma Deep Raj , Gill Kiran Dip
Published: December 2012

Dichlorvos, an organophosphate (OP), is known to cause oxidative stress in the central nervous system (CNS). Previously we have shown that dichlorvos treatment promoted the levels of proinflammatory molecules and ultimately induced apoptotic cell death in primary microglial cells. Here we studied the effect of dichlorvos on crucial cell cycle regulatory proteins and the DNA damage sensor ataxia‐telangiectasia mutated (ATM). We found a significant increase in p53 and its downstream target, p21, levels in dichlorvos‐treated microglial cells compared with control cells. Moreover, dichlorvos exposure promoted the levels of different cell cycle regulatory proteins. These results along with flow cytometry results suggested that primary microglial cells were arrested at G1 and G2/M phase after dichlorvos exposure. We have shown in a previous study that dichlorvos can induce DNA damage in microglia; here we found that microglial cells also tried to repair this damage by inducing a DNA repair enzyme, i.e., ATM. We observed a significant increase in the levels of ATM after dichlorvos treatment compared with control. © 2012 Wiley Periodicals, Inc.
Abbreviated exposure to cuprizone is sufficient to induce demyelination and oligodendrocyte loss
Authors: Doan Vivian , Kleindienst Amber M. , McMahon Eileen J. , Long Brian R. , Matsushima Glenn K. , Taylor Lorelei C.
Published: December 2012

Cuprizone intoxication is one of several animal models used to study demyelination and remyelination. Early treatment protocols exposed mice to cuprizone for 6 weeks to induce demyelination; however, more recent reports have varied exposure times from 4 to 5 weeks. The goal of this study was to determine the minimal exposure of cuprizone in C57BL/6 mice that would induce a pathology of robust demyelination and gliosis similar to that described for a 5‐ or 6‐week treatment. We found that an abbreviated insult of only 2 weeks of exposure to cuprizone induced significant demyelination 3 weeks later (5‐week time point) but was somewhat variable. Three weeks of exposure to cuprizone produced extensive demyelination by week 5, equivalent to that observed with 5 weeks of exposure. The depletion of mature oligodendrocytes, as well as microglia and astrocyte accumulation, showed trends similar to those with 5‐week exposure to cuprizone. Once mature oligodendrocytes are perturbed after a 3‐week treatment, the progression to demyelination occurs without requiring further exposure. Furthermore, the early removal of cuprizone did not accelerate remyelination, suggesting that other sequences of events must follow before repair can occur. Thus, a short, “hit and run” CNS insult triggers a cascade of events leading to demyelination 2–3 weeks later. © 2012 Wiley Periodicals, Inc.
Electronic magnification and perceived contrast of video
Authors: Haun Andrew M. , Peli Eli , Woods Russell L.
Published: December 2012

Electronic magnification of an image results in a decrease in its perceived contrast. The decrease in perceived contrast could be due to a perceived blur or to limited sampling of the range of contrasts in the original image. We measured the effect on perceived contrast of magnification in two contexts: either a small video was enlarged to fill a larger area or a portion of a larger video was enlarged to fill the same area as the original. Subjects attenuated the source video contrast to match the perceived contrast of the magnified videos, with the effect increasing with magnification and decreasing with viewing distance. These effects are consistent with expectations based on both the contrast statistics of natural images and the contrast sensitivity of the human visual system. We demonstrate that local regions within videos usually have lower physical contrast than the whole and that this difference accounts for a minor part of the perceived differences. Instead, visibility of “missing content” (blur) in a video is misinterpreted as a decrease in contrast. We detail how the effects of magnification on perceived contrast can be measured while avoiding confounding factors.
Removing the ambiguity of data processing methods: Optimizing the location of peak boundaries for accurate moment calculations
Authors: Stevenson Paul G. , Gao Hong , Gritti Fabrice , Guiochon Georges
Published: December 2012

The calculation of the first few moments of elution peaks is necessary to determine: the amount of component in the sample (peak area or zeroth moment), the retention factor (first moment), and the column efficiency (second moment). It is a time consuming and tedious task for the analyst to perform these calculations, thus data analysis is generally completed by data stations associated to modern chromatographs. However, data acquisition software is a black box which provides no information to chromatographers on how their data are treated. These results are too important to be accepted on blind faith. The location of the peak integration boundaries is most important. In this manuscript, we explore the relationships between the size of the integration area, the relative position of the peak maximum within this area, and the accuracy of the calculated moments. We found that relationships between these parameters do exist and that computers can be programmed with relatively simple routines to automatize the extraction of key peak parameters and to select acceptable integration boundaries. It was also found that the most accurate results are obtained when the S/N exceeds 200.
A rapid and simple separation and direct detection of glutathione by gold nanoparticles and graphene‐based MALDI–TOF‐MS
Authors: Wan Dan , Gao Mingxia , Wang Yuhua , Zhang Peng , Zhang Xiangmin
Published: December 2012

In this study, we present a rapid and simple method for the separation and direct detection of glutathione by combining gold nanoparticles and MALDI–TOF‐MS with graphene as matrix. Gold nanoparticles enable the selective capture of thiol‐containing compounds. Gold nanoparticles bound with analytes can be mixed with graphene matrix for direct analysis by MALDI–TOF‐MS, which can avoid sample loss and contamination during transfer process. Compared with a conventional matrix, α‐cyano‐4‐hydroxycinnamic acid, graphene exhibits an excellent desorption/ionization efficiency, thermal and mechanical properties. The use of graphene as matrix avoids the fragmentation of analytes. Stable analysis was achieved with less background interference even at the concentration of 0.625 ng/μL. To further confirm its efficiency, the optimized approach was applied to the separation and detection of glutathione in mouse liver extraction. This result showed the great potential of detection of biologically important thiols in biochemical and biomedical research.