bluuebunny/crossref_metadata_2025 huggingface dataset:
Solid‐Phase Synthesis of Oligonucleotide 5′‐(α‐P‐Thio)triphosphates and 5′‐(α‐P‐Thio)(β,γ‐methylene)triphosphates
Authors: Thillier Yann , Sallamand Corinne , Baraguey Carine , Vasseur Jean‐Jacques , Debart Françoise
Published: December 2014

A robust solid‐phase synthesis was developed to obtain original oligonucleotides (ONs) functionalized at their 5′ end with modified triphosphate (TP) moieties, in which a nonbridging oxygen atom of the α phosphorus atom was replaced by a sulfur atom and the labile P–O–P linkage was changed into a methylene bridge between the β and γ phosphorus atoms. The efficient method is based on solid‐supported ON assembly followed by 5′‐H‐phosphonylation, oxidation to the thiophosphate subsequently activated as a phosphoanhydride with diphenyl phosphoryl chloride, then nucleophilic substitution with the alkylammonium salt of pyrophosphate or its β,γ‐methylene analogue. After deprotection and release from the solid support under basic conditions, 5′‐(α‐P‐thio)TP and 5′‐(α‐P‐thio)(β,γ‐methylene)TP oligonucleotides were obtained in satisfactory yields, and they were isolated with high purity. These hydrolysis‐resistant 5′‐TP ONs will be useful in biological research to elucidate the mechanism of enzymes involved in mRNA processing and maturation.
Presence and predictors of persistent pain among persons who sustained an injury in a road traffic crash
Authors: Gopinath B. , Jagnoor J. , Nicholas M. , Blyth F. , Harris I.A. , Casey P. , Cameron I.D.
Published: December 2014

Background

There is a paucity of prospective studies with long follow‐up that have examined a wide range of correlates associated with persistent pain outcomes in persons who sustained a mild or moderate injury in a road traffic crash. This study aimed to establish the independent predictors of pain severity over 24 months.

Methods

A total of 364, 284 and 252 persons with mild/moderate musculoskeletal injuries sustained in a vehicle‐related crash participated in telephone interviews in the subacute phase, and at 12 and 24 months, respectively. The numeric rating scale (NRS) assessed pain severity. Pain‐Related Self‐Statements Scale‐Catastrophizing (PRSS‐Catastrophizing) and the Short Form Orebro Musculoskeletal Pain Screening Questionnaire (OMPSQ) were also administered.

Results

After multivariable adjustment, each 1 SD increase in Short Form‐12 Physical Component Score (SF‐12 PCS) in the subacute phase was associated with 0.73 (p = 0.002) and 1.11 (p < 0.0001) decrease in NRS scores after 12 and 24 months, respectively. Each unit increase in the PRSS‐Catastrophizing score in the subacute phase was associated with 0.54 (p = 0.001) and 0.43 (p = 0.03) increase in NRS scores 12 and 24 months later, respectively. Subacute phase OMPSQ scores were positively associated with NRS scores at 12‐ and 24‐month follow‐ups (p < 0.0001).

Conclusions

Self‐perceived physical well‐being, pain‐related work disability and pain catastrophizing could play a role in determining long‐term pain‐related outcomes following traffic‐related injuries.
Optimization of Os‐Complex Modified Redox Polymers for Improving Biocatalysis of PQQ‐sGDH Based Electrodes
Authors: Pinyou Piyanut , Pöller Sascha , Chen Xingxing , Schuhmann Wolfgang
Published: December 2014

A variety of Os‐complex modified redox polymers were synthesized and optimized for an improved bioelectrochemical communication with PQQ‐sGDH. The properties of the polymers were varied by changing the monomers composition leading to improved immobilization and increased enzyme loading. Three Os‐complexes were designed exhibiting different ligands but similar redox potentials adapted for efficient electron transfer with PQQ‐sGDH. Effects of the ligand sphere, polymer backbone and length of the tether chain between Os‐complex and polymer backbone as well as the impact of different bifunctional crosslinkers on the catalytic current were evaluated. The optimized bioelectrodes exhibited significantly improved catalytic currents and stability.
On‐Site Determination of Carbendazim, Cathecol and Hydroquinone in Tap Water Using a Homemade Batch Injection Analysis Cell for Screen Printed Electrodes
Authors: Caramit Ricardo Pini , Lucca Bruno Gabriel , Souza Ferreira Valdir , Abarza Munoz Rodrigo Alejandro , Richter Eduardo Mathias , da Silva Rodrigo Amorim Bezerra
Published: December 2014

In this work, we present a simple homemade batch‐injection analysis cell for screen‐printed electrodes (BIA‐SPE). The potential of the proposed system for on‐site analysis was demonstrated by the determination of carbendazim, catechol, and hydroquinone in tap water. The system provided reduced injection volume (30 µL), high analytical frequency (≈200 h−1) and low detection limits (nanomolar level). Moreover, the BIA‐SPE cell presented better stability (RSD≈0.4 %) than a conventional flow injection cell for SPE (RSD≈5.0 %) in organic media. The proposed homemade BIA‐SPE cell is very simple, inexpensive and can be easily constructed in any laboratory.
A Solid Film Electrode of Intermetallic Ag2Hg3.02 for the Direct Determination of Folic Acid in Fresh and Processed Fruit Juices
Authors: Garcia de Paula Cleide , Alves de Toledo Renata , Vaz Carlos Manoel Pedro , Marques Cordeiro Paulo Jorge , Batistuti José Paschoal , Aparecida Pastre Ieda , Shim Hojae , Fertonani Fernando Luís
Published: December 2014

AgSIE was used for the direct analysis of folic acid (FA), with a detection limit and lower level of quantitation of 6.8×10−10 mol L−1 and 2.3×10−8 mol L−1. The analysis in fresh and processed fruits was done without any sample pretreatment. In strawberry and acerola juices, FA concentration level values were below the method detection limit. FA was detectable in peach (77.7±0.4 µg L−1 and 64.4±0.5 µg L−1), Persian lime (45.4±0.7 µg L−1), pineapple Hawaii (66.2±0.4 µg L−1), pear pineapple (35.3±0.6 µg L−1), cashew (54.4±0.5 µg L−1), passion fruit (73.2±0.3 µg L−1), and apple (84.4±0.5 µg L−1).
High‐throughput DNA separation in nanofilter arrays
Authors: Choi Sungup , Kim Ju Min , Ahn Kyung Hyun , Lee Seung Jong
Published: December 2014

Electrophoresis 2014, 35, 2068–2077. DOI: 10.1002/elps.elps201400168

The non‐uniform field effect combined with geometric constraint plays a key role in nanofilter‐based DNA separation. DNA molecules in nanofilter arrays deterministically move along different electrophoretic streamlines according to their length, which is the origin of high‐throughput DNA separation.

image
Expanding metal mixture toxicity models to natural stream and lake invertebrate communities
Authors: Balistrieri Laurie S. , Mebane Christopher A. , Schmidt Travis S. , Keller Wendel (Bill)
Published: December 2014

A modeling approach that was used to predict the toxicity of dissolved single and multiple metals to trout is extended to stream benthic macroinvertebrates, freshwater zooplankton, and Daphnia magna. The approach predicts the accumulation of toxicants (H, Al, Cd, Cu, Ni, Pb, and Zn) in organisms using 3 equilibrium accumulation models that define interactions between dissolved cations and biological receptors (biotic ligands). These models differ in the structure of the receptors and include a 2-site biotic ligand model, a bidentate biotic ligand or 2-pKa model, and a humic acid model. The predicted accumulation of toxicants is weighted using toxicant-specific coefficients and incorporated into a toxicity function called Tox, which is then related to observed mortality or invertebrate community richness using a logistic equation. All accumulation models provide reasonable fits to metal concentrations in tissue samples of stream invertebrates. Despite the good fits, distinct differences in the magnitude of toxicant accumulation and biotic ligand speciation exist among the models for a given solution composition. However, predicted biological responses are similar among the models because there are interdependencies among model parameters in the accumulation–Tox models. To illustrate potential applications of the approaches, the 3 accumulation–Tox models for natural stream invertebrates are used in Monte Carlo simulations to predict the probability of adverse impacts in catchments of differing geology in central Colorado (USA); to link geology, water chemistry, and biological response; and to demonstrate how this approach can be used to screen for potential risks associated with resource development. Environ Toxicol Chem 2015;34:761–776. © 2015 SETAC
Estimation of herbicide species sensitivity distribution using single-species algal toxicity data and information on the mode of action
Authors: Nagai Takashi , Taya Kiyoshi
Published: December 2014

Although species sensitivity distribution (SSD) is a key concept for quantitative ecological risk assessment, its application is limited owing to a lack of sufficient data for the analysis, especially on the toxicity of herbicides for primary producers. The authors developed a method of herbicide SSD estimation using single-species toxicity data and information on the herbicide mode of action. The authors' method was based on 2 assumptions: the slopes of the SSD of the same MOA herbicides are the same and the relative sensitivities of standard algae in the SSD of the same MOA herbicides are the same. The 2 parameters of log-normal SSD, mean sensitivity, and variation in sensitivity, for 92 herbicides were determined to establish the estimation model. Mean sensitivities were linearly correlated with logarithmic 50% effect concentrations (EC50) for standard algae. The average of variations in sensitivity significantly differed among MOA, and variations in sensitivity were constant independently of EC50 values for standard algae for the same MOA herbicides. These results were all consistent with the assumptions of the SSD estimation method. The outcome was validated by comparing the estimated SSDs using the proposed method with the generated SSDs using toxicity data which were independent of method development. These SSDs were consistent, and considering MOA information improved the accuracy of estimating SSD markedly. Environ Toxicol Chem 2015;34:677–684. © 2014 SETAC
Application of POCIS for exposure assessment of munitions constituents during constant and fluctuating exposure
Authors: Belden Jason B. , Lotufo Guilherme R. , Biedenbach James M. , Sieve Kristal K. , Rosen Gunther
Published: December 2014

The present study examined the potential use of polar organic chemical integrative samplers (POCIS) for exposure assessment of munitions constituents, including 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), and their breakdown products (aminodinitrotoluenes [ADNTs], diaminonitrotoluenes [DANTs], and hexahydro-1,3,5-trinitroso-1,3,5-triazine [TNX]). Loss of munitions constituents from the sorbent phase after uptake was observed for the “pesticide” POCIS configuration but not for the “pharmaceutical” configuration. Therefore, the latter was selected for further investigation. Under constant exposure conditions, TNT, ADNTs, DANT, RDX, and atrazine (a common environmental contaminant) accumulated at a linear rate for at least 14 d, with sampling rates between 34 mL/d and 215 mL/d. When POCIS were exposed to fluctuating concentrations, analyte accumulation values were similar to values found during constant exposure, indicating that the sampler was indeed integrative. In contrast, caffeine (a common polar contaminant) and TNX did not accumulate at a linear rate and had a reduction in accumulation of greater than 50% on the POCIS during fluctuating exposures, demonstrating that POCIS did not sample those chemicals in an integrative manner. Moreover, in a flow-through microcosm containing the explosive formulation Composition B, TNT and RDX were readily measured using POCIS, despite relatively high turnover rates and thus reduced water concentrations. Mean water concentrations estimated from POCIS were ± 37% of mean water concentrations measured by traditional grab sample collection. Thus, POCIS were found to have high utility for quantifying exposure to most munitions constituents evaluated (TNT, ADNTs, and RDX) and atrazine. Environ Toxicol Chem 2015;34:959–967. © 2014 SETAC
Metal Mixture Modeling Evaluation project: 3. Lessons learned and steps forward
Authors: Farley Kevin J. , Meyer Joseph S.
Published: December 2014

A comparison of 4 metal mixture toxicity models (that were based on the biotic ligand model [BLM] and the Windermere humic aqueous model using the toxicity function [WHAM-FTOX]) was presented in a previous paper. In the present study, a streamlined version of the 4 models was developed and applied to multiple data sets and test conditions to examine key assumptions and calibration strategies that are crucial in modeling metal mixture toxicity. Results show that 1) a single binding site on or in the organism was a useful and oftentimes sufficient framework for predicting metal toxicity; 2) a linear free energy relationship (LFER) for bidentate binding of metals and cations to the biotic ligand provided a good first estimate of binding coefficients; 3) although adjustments in metal binding coefficients or adjustments in chemical potency factors can both be used in model calibration for single-metal exposures, changing metal binding coefficients or chemical potency factors had different effects on model predictions for metal mixtures; and 4) selection of a mixture toxicity model (based on concentration addition or independent action) was important in predicting metal mixture toxicity. Moving forward, efforts should focus on reducing uncertainties in model calibration, including development of better methods to characterize metal binding to toxicologically active binding sites, conducting targeted exposure studies to advance the understanding of metal mixture toxicity, and further developing LFERs and other tools to help constrain the model calibration. Environ Toxicol Chem 2015;34:821–832. © 2014 SETAC
SDN‐based 5G mobile networks: architecture, functions, procedures and backward compatibility
Authors: Trivisonno R. , Guerzoni R. , Vaishnavi I. , Soldani D.
Published: December 2014

In this paper, we describe an SDN‐based plastic architecture for 5G networks, designed to fulfill functional and performance requirements of new generation services and devices. The 5G logical architecture is presented in detail, and key procedures for dynamic control plane instantiation, device attachment, and service request and mobility management are specified. Key feature of the proposed architecture is flexibility, needed to support efficiently a heterogeneous set of services, including Machine Type Communication, Vehicle to X and Internet of Things traffic. These applications are imposing challenging targets, in terms of end‐to‐end latency, dependability, reliability and scalability. Additionally, backward compatibility with legacy systems is guaranteed by the proposed solution, and Control Plane and Data Plane are fully decoupled. The three levels of unified signaling unify Access, Non‐access and Management strata, and a clean‐slate forwarding layer, designed according to the software defined networking principle, replaces tunneling protocols for carrier grade mobility. Copyright © 2014 John Wiley & Sons, Ltd.
Karyoevelution on the genus Onosma L. (Boraginaceae): quantification of chromosome number heterogeneity
Authors: Almasi Maryam , Ranjbar Massoud
Published: December 2014

In this survey, chromosome counts of different species belonging to the genus Onosma are summarized and then karyological patterns available including frequency of cytotype occurrence, percentage of taxa with particular basic chromosome number and rate of polyploidy in the genus are evaluated. Quantitative parameters have been used to characterize chromosome number (CN) variation. In order to verify if variation patterns differ between three groups of Onosma, Index of CN Heterogeneity (ICNH) was quantified. In addition, meiotic chromosome numbers of 14 populations belonging to 11 species growing in Iran, namely Onosma araratica (2n = 2x = 16), O. asperrima (2n = 2x = 16), O. bulbotricha (2n = 2x = 18), O. kotschyi (2n = 2x = 16), O. microcarpa (2n = 2x = 16), O. nigricaulis (2n = 2x = 16), O. nervosa (2n = 2x = 16), O. obtusifolia (2n = 2x = 16), O. pachypoda (2n = 2x = 16), O. stenosiphon (2n = 2x = 20) and O. subsericea (2n = 2x = 16), were determined. With the exception of O. microcarpa and O. bulbotricha, all chromosome counts are reported for the first time. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
A high‐order discontinuous Galerkin method for all‐speed flows
Authors: Renda S. M. , Hartmann R. , De Bartolo C. , Wallraff M.
Published: December 2014

In this article, we present a discontinuous Galerkin (DG) method designed to improve the accuracy and efficiency of steady solutions of the compressible fully coupled Reynolds‐averaged Navier–Stokes and k − ω turbulence model equations for solving all‐speed flows. The system of equations is iterated to steady state by means of an implicit scheme. The DG solution is extended to the incompressible limit by implementing a low Mach number preconditioning technique. A full preconditioning approach is adopted, which modifies both the unsteady terms of the governing equations and the dissipative term of the numerical flux function by means of a new preconditioner, on the basis of a modified version of Turkel's preconditioning matrix. At sonic speed the preconditioner reduces to the identity matrix thus recovering the non‐preconditioned DG discretization. An artificial viscosity term is added to the DG discretized equations to stabilize the solution in the presence of shocks when piecewise approximations of order of accuracy higher than 1 are used. Moreover, several rescaling techniques are implemented in order to overcome ill‐conditioning problems that, in addition to the low Mach number stiffness, can limit the performance of the flow solver. These approaches, through a proper manipulation of the governing equations, reduce unbalances between residuals as a result of the dependence on the size of elements in the computational mesh and because of the inherent differences between turbulent and mean‐flow variables, influencing both the evolution of the Courant Friedrichs Lewy (CFL) number and the inexact solution of the linear systems. The performance of the method is demonstrated by solving three turbulent aerodynamic test cases: the flat plate, the L1T2 high‐lift configuration and the RAE2822 airfoil (Case 9). The computations are performed at different Mach numbers using various degrees of polynomial approximations to analyze the influence of the proposed numerical strategies on the accuracy, efficiency and robustness of a high‐order DG solver at different flow regimes. Copyright © 2014 John Wiley & Sons, Ltd.
Influences of a DRD2 polymorphism on updating of long‐term memory representations and caudate BOLD activity: Magnification in aging
Authors: Persson Jonas , Rieckmann Anna , Kalpouzos Grégoria , Fischer Håkan , Bäckman Lars
Published: December 2014

A number of genetic polymorphisms are related to individual differences in cognitive performance. Striatal dopamine (DA) functions, associated with cognitive performance, are linked to the TaqIA polymorphism of the DRD2/ANKK1 gene. In humans, presence of an A1 allele of the DRD2/ANKK1‐TaqIA polymorphism is related to reduced density of striatal DA D2 receptors. The resource‐modulation hypothesis assumes that aging‐related losses of neurochemical and structural brain resources modulate the extent to which genetic variations affect cognitive functioning. Here, we tested this hypothesis using functional MRI during long‐term memory (LTM) updating in younger and older carriers and noncarriers of the A1‐allele of the TaqIa polymorphism. We demonstrate that older A1‐carriers have worse memory performance, specifically during LTM updating, compared to noncarriers. Moreover, A1‐carriers exhibited less blood oxygen level‐dependent (BOLD) activation in left caudate nucleus, a region critical to updating. This effect was only seen in older adults, suggesting magnification of genetic effects on functional brain activity in aging. Further, a positive relationship between caudate BOLD activation and updating performance among non‐A1 carriers indicated that caudate activation was behaviorally relevant. These results demonstrate a link between the DRD2/ANKK1‐TaqIA polymorphism and neurocognitive deficits related to LTM updating, and provide novel evidence that this effect is magnified in aging. Hum Brain Mapp 36:1325–1334, 2015. © 2014 Wiley Periodicals, Inc.
Submillisecond unmasked subliminal visual stimuli evoke electrical brain responses
Authors: Sperdin Holger F. , Spierer Lucas , Becker Robert , Michel Christoph M. , Landis Theodor
Published: December 2014

Subliminal perception is strongly associated to the processing of meaningful or emotional information and has mostly been studied using visual masking. In this study, we used high density 256‐channel EEG coupled with an liquid crystal display (LCD) tachistoscope to characterize the spatio‐temporal dynamics of the brain response to visual checkerboard stimuli (Experiment 1) or blank stimuli (Experiment 2) presented without a mask for 1 ms (visible), 500 µs (partially visible), and 250 µs (subliminal) by applying time‐wise, assumption‐free nonparametric randomization statistics on the strength and on the topography of high‐density scalp‐recorded electric field. Stimulus visibility was assessed in a third separate behavioral experiment. Results revealed that unmasked checkerboards presented subliminally for 250 µs evoked weak but detectable visual evoked potential (VEP) responses. When the checkerboards were replaced by blank stimuli, there was no evidence for the presence of an evoked response anymore. Furthermore, the checkerboard VEPs were modulated topographically between 243 and 296 ms post‐stimulus onset as a function of stimulus duration, indicative of the engagement of distinct configuration of active brain networks. A distributed electrical source analysis localized this modulation within the right superior parietal lobule near the precuneus. These results show the presence of a brain response to submillisecond unmasked subliminal visual stimuli independently of their emotional saliency or meaningfulness and opens an avenue for new investigations of subliminal stimulation without using visual masking. Hum Brain Mapp 36:1470–1483, 2015. © 2014 Wiley Periodicals, Inc.
Adenoid cystic carcinoma: A review of recent advances, molecular targets, and clinical trials
Authors: Dillon Patrick M. , Chakraborty Samhita , Moskaluk Christopher A. , Joshi Prashant J. , Thomas Christopher Y.
Published: December 2014

Background

Adenoid cystic carcinoma (ACC) is a rare tumor of secretory glands. In this study, recent advances in molecular characterization and in therapeutics are reviewed.

Methods

A search of articles in PubMed and of abstracts from national meetings was performed regarding ACC.

Results

Recent genetic analyses found that recurrent chromosome 6:9 translocations in ACC generate an MYB:NFIB gene fusion resulting in overexpression of the MYB oncoprotein. Several other frequent mutations are recently published that may be relevant for drug development. Several trials of targeted drugs are reviewed. Some agents delay tumor progression, but tumor responses remain rare.

Conclusion

ACCs have a characteristic chromosomal translocation, but also frequently pick up additional mutations. Clinical research is limited by the rarity and slow growth of ACC. Several ongoing trials are testing agents that inhibit fibroblast growth factor receptor signaling or other signaling pathways. Novel treatments based on the recently sequenced tumor genome are under development. © 2015 The Authors Head & Neck Published by Wiley Periodicals, Inc. 38: 620–627, 2016
Efficient, effective, safe procedure to identify nonrecurrent inferior laryngeal nerve during thyroid surgery
Authors: Watanabe Akihito , Taniguchi Masanobu , Kimura Yuki , Ito Suguru , Hosokawa Masao , Sasaki Shigeyuki
Published: December 2014

Background

The nonrecurrent inferior laryngeal nerve (NRILN) is always associated with the aberrant subclavian artery. CT images can detect this vascular anomaly, which predicts an NRILN. The purpose of this study was to report our procedure to identify the NRILN in patients with the aberrant subclavian artery.

Methods

Four of 730 patients undergoing thyroid operation in our hospital were preoperatively diagnosed with aberrant subclavian artery by CT of the neck. To avoid vocal cord paralysis, we approached the vagal nerve first before dissecting the paratracheal region to discover the separation point of the NRILN from the vagal nerve.

Results

The NRILN was identified without difficulty in all 4 patients. No patients showed vocal cord paralysis.

Conclusion

Approaching the vagal nerve first before dissecting the paratracheal region is an efficient, effective, and safe procedure to identify an NRILN in patients who are preoperatively diagnosed as having the aberrant subclavian artery. © 2015 Wiley Periodicals, Inc. Head Neck 38: 573–577, 2016
Extent of neck dissection after transoral robotic surgical resection of oropharyngeal squamous cell carcinoma: Report of a case and potential indications for inclusion of level I in a selective neck dissection
Authors: Noel Christopher W. , Foreman Andrew , Goldstein David P. , de Almeida John R.
Published: December 2014

Background

With new minimally invasive surgical techniques playing a role in the management of early oropharyngeal carcinomas, appropriate surgical management of the neck is an increasingly important consideration.

Methods

We present a case of a 47‐year‐old patient with a clinical T2N1M0 squamous cell carcinoma (SCC) of the right tonsil with anterior extension toward the floor of the mouth from the right glossotonsillar sulcus. Imaging revealed a solitary necrotic lymph node in level IIA. The decision was made to proceed with a transoral robotic pharyngectomy and selective neck dissection of levels II to IV, followed by adjuvant radiotherapy.

Results

A postoperative scan revealed progressive regional disease in the anterior aspect of level IB, and, as such, a completion neck dissection of level I was performed.

Conclusion

Special consideration must be given to level I, particularly in patients with glossotonsillar sulcus involvement and anterior extension toward the floor of the mouth. © 2015 Wiley Periodicals, Inc. Head Neck 37: E130–E133, 2015
Characterization of cytoplasmic polyadenylation element binding 2 protein expression and its RNA binding activity
Authors: Turimella Sada Lakshmi , Bedner Peter , Skubal Magdalena , Vangoor Vamshidhar Reddy , Kaczmarczyk Lech , Karl Kevin , Zoidl Georg , Gieselmann Volkmar , Seifert Gerald , Steinhäuser Christian , Kandel Eric , Theis Martin
Published: December 2014

Cytoplasmic polyadenylation element binding (CPEB) proteins are translational regulators that are involved in the control of cellular senescence, synaptic plasticity, learning, and memory. We have previously found all four known CPEB family members to be transcribed in the mouse hippocampus. Aside from a brief description of CPEB2 in mouse brain, not much is known about its biological role. Hence, this study aims to investigate CPEB2 expression in mouse brain. With reverse transcription polymerase chain reaction (RT‐PCR) of total mouse brain cDNA, we identified four distinct CPEB2 splice variants. Single‐cell RT‐PCR showed that CPEB2 is predominantly expressed in neurons of the juvenile and adult brain and that individual cells express different sets of splice variants. Staining of brain slices with a custom‐made CPEB2 antibody revealed ubiquitous expression of the protein in many brain regions, including hippocampus, striatum, thalamus, cortex, and cerebellum. We also found differential expression of CPEB2 protein in excitatory, inhibitory, and dopaminergic neurons. In primary hippocampal cultures, the subcellular localization of CPEB2 in neurons and astrocytes resembled that of CPEB1. Electrophoretic mobility shift assay and RNA coimmunoprecipitation revealed CPEB2 interaction with β‐catenin and Ca2+/calmodulin‐dependent protein kinase II (both established CPEB1 targets), indicating an overlap in RNA binding specificity between CPEB1 and CPEB2. Furthermore, we identified ephrin receptor A4 as a putative novel target of CPEB2. In conclusion, our study identifies CPEB2 splice variants to be differentially expressed among individual cells and across cell types of the mouse hippocampus, and reveals overlapping binding specificity between CPEB2 and CPEB1. © 2014 Wiley Periodicals, Inc.
Flow paths and phosphorus transfer pathways in two agricultural streams with contrasting flow controls
Authors: Mellander Per‐Erik , Jordan Phil , Shore Mairead , Melland Alice R. , Shortle Ger
Published: December 2014

In this paper, we analyse 4 years of data from simultaneous high‐frequency monitoring of streamflow and phosphorus (P) concentration. This was carried out to investigate hydrological flow paths and P transfer pathways from diffuse sources in two intensively farmed river catchments (~10 km2) with contrasting flow controls and dominating flow paths. Catchment scale P loss was viewed on an annual and event flow basis and related to hydrological flow paths. A grassland catchment with mostly poorly drained soils, and a higher Q10:Q90 ratio (60 compared with 24), had three times higher annual P loss than an arable catchment with mostly well‐drained soils (1.04 compared with 0.34 kg TP ha−1) despite the arable catchment having larger areas with high soil P status and more discharge. Neither of the catchments indicated supply limitations. The magnitude of the P losses from the two catchments was not defined by land use, source pressure or discharge volume but rather by more basic rainfall‐to‐runoff partitioning influences that determine proportions of quickflow and slowflow. There were larger differences between the years than between the catchments, and the P loss of the arable catchment appeared more sensitive to climate. The results confirmed the need to manage the quickflow components of runoff to moderate P transfers. Therefore, in order to further reduce diffuse pollution it may be necessary to account for the contrast in hydrological function before or in addition to any of the other factors known to influence P losses from catchments (such as soil P and land use). Schemes designed to attenuate diffuse P after mobilization from soil surfaces can then be targeted (and resourced) more effectively. Copyright © 2014 John Wiley & Sons, Ltd.
HMGB1 secretion during cervical carcinogenesis promotes the acquisition of a tolerogenic functionality by plasmacytoid dendritic cells
Authors: Demoulin Stéphanie , Herfs Michael , Somja Joan , Roncarati Patrick , Delvenne Philippe , Hubert Pascale
Published: December 2014

Acquisition of an impaired functionality by plasmacytoid dendritic cells (pDCs) contributing to cancer progression has been documented in different types of cancers. In the present study, we postulate that molecules secreted by (pre)neoplastic epithelial cells of the genital tract (cervix/vulva) might attract pDCs but also modify their proper functionality, allowing these cells to initiate a tolerogenic response interfering with antitumor immunity. We demonstrated that pDCs are recruited during the cervical metaplasia‐dysplasia‐cancer sequence, through the action of their chemoattractant, chemerin. We showed that stimulated‐pDCs exposed to cervical/vulvar tumor microenvironment display an altered phenotype. We also demonstrated that cervical/vulvar neoplastic keratinocytes inhibit the proper function of pDCs by decreasing their IFNα secretion in response to CpG oligonucleotides. In parallel, we observed that (pre)neoplastic areas of the cervix are infiltrated by FoxP3+ Treg cells which colocalize with pDCs. Accordingly, pDCs cocultured with cervical/vulvar neoplastic keratinocytes have the capacity to induce a Treg cell differentiation from naïve CD4+ T cells, which is in agreement with the development of a tolerogenic response. We identified HMGB1 as a soluble factor produced by neoplastic keratinocytes from the genital tract involved in pDCs functional alteration. Indeed, this molecule inhibited pDC maturation, decreased IFNα secretion following TLR9 stimulation and forced these cells to become tolerogenic. In contrast, inhibition of HMGB1 restored pDC phenotype. Our findings indicate that the use of inhibitory molecules notably directed against HMGB1 in cervical/vulvar (pre)neoplastic lesions might prevent alterations of pDCs functionality and represent an attractive therapeutic strategy to overcome immune tolerance in cancers.
Quick analysis of optical spectra to quantify epidermal melanin and papillary dermal blood content of skin
Authors: Jacques Steven L.
Published: December 2014

This paper presents a practical approach for assessing the melanin and blood content of the skin from total diffuse reflectance spectra, R(λ), where λ is wavelength. A quick spectral analysis using just three wavelengths (585 nm, 700 nm and 800 nm) is presented, based on the 1985 work of Kollias and Baquer who documented epidermal melanin of skin using the slope of optical density (OD) between 620 nm and 720 nm. The paper describes the non‐rectilinear character of such a quick analysis, and shows that almost any choice of two wavelengths in the 600–900 range can achieve the characterization of melanin. The extrapolation of the melanin slope to 585 nm serves as a baseline for subtraction from the OD (585 nm) to yield a blood perfusion score. Monte Carlo simulations created spectral data for a skin model with epidermis, papillary dermis and reticular dermis to illustrate the analysis.

magnified image
Macrophages Promote Osteoblastic Differentiation In Vivo: Implications in Fracture Repair and Bone Homeostasis
Authors: Vi Linda , Baht Gurpreet S , Whetstone Heather , Ng Adeline , Wei Qingxia , Poon Raymond , Mylvaganam Sivakami , Grynpas Marc , Alman Benjamin A
Published: December 2014

Macrophages are activated in inflammation and during early phases of repair processes. Interestingly, they are also present in bone during development, but their function during this process is unclear. Here, we explore the function of macrophages in bone development, growth, and repair using transgenic mice to constitutively or conditionally deplete macrophages. Depletion of macrophages led to early skeletal growth retardation and progressive osteoporosis. By 3 months of age, macrophage-deficient mice displayed a 25% reduction in bone mineral density and a 70% reduction in the number of trabecular bone compared to control littermates. Despite depletion of macrophages, functional osteoclasts were still present in bones, lining trabecular bone and the endosteal surface of the cortical bone. Furthermore, ablation of macrophages led to a 60% reduction in the number of bone marrow mesenchymal progenitor cells and a decrease in the ability of these cells to differentiate to osteoblasts. When macrophages were depleted during fracture repair, bone union was impaired. Calluses from macrophage-deficient animals were smaller, and contained less bone and more fibrotic tissue deposition. Taken together, this shows that macrophages are crucial for maintaining bone homeostasis and promoting fracture repair by enhancing the differentiation of mesenchymal progenitors. © 2014 American Society for Bone and Mineral Research.
n‐Octyl Gallate as Inhibitor of Pyruvate Carboxylation and Lactate Gluconeogenesis
Authors: Eler Gabrielle Jacklin , Santos Israel Souza , de Moraes Amarilis Giaretta , Comar Jurandir Fernando , Peralta Rosane Marina , Bracht Adelar
Published: December 2014

The alkyl gallates are found in several natural and industrial products. In the latter products, these compounds are added mainly for preventing oxidation. In the present work, the potencies of methyl gallate, n‐propyl gallate, n‐pentyl gallate, and n‐octyl gallate as inhibitors of pyruvate carboxylation and lactate gluconeogenesis were evaluated. Experiments were done with isolated mitochondria and the isolated perfused rat liver. The potency of the gallic acid esters as inhibitors of pyruvate carboxylation in isolated mitochondria obeyed the following decreasing sequence: n‐octyl gallate > n‐pentyl gallate > n‐propyl gallate > methyl gallate. A similar sequence of decreasing potency for lactate gluconeogenesis inhibition in the perfused liver was found in terms of the portal venous concentration. Both actions correlate with the lipophilicity of the compounds. The effects are harmful at high concentrations. At appropriate concentrations, however, octyl gallate should act therapeutically because its inhibitory action on gluconeogenesis will contribute further to its proposed antihyperglycemic effects.
Automatic algorithms for completeness‐optimization of Gaussian basis sets
Authors: Lehtola Susi
Published: December 2014

We present the generic, object‐oriented C++ implementation of the completeness‐optimization approach (Manninen and Vaara, J. Comput. Chem. 2006, 27, 434) in the freely available ERKALE program, and recommend the addition of basis set stability scans to the completeness‐optimization procedure. The design of the algorithms is independent of the studied property, the used level of theory, as well as of the role of the optimized basis set: the procedure can be used to form auxiliary basis sets in a similar fashion. This implementation can easily be interfaced with various computer programs for the actual calculation of molecular properties for the optimization, and the calculations can be trivially parallelized. Routines for general and segmented contraction of the generated basis sets are also included. The algorithms are demonstrated for two properties of the argon atom—the total energy and the nuclear magnetic shielding constant—and they will be used in upcoming work for generation of cost‐efficient basis sets for various properties. © 2014 Wiley Periodicals, Inc.
pmx: Automated protein structure and topology generation for alchemical perturbations
Authors: Gapsys Vytautas , Michielssens Servaas , Seeliger Daniel , de Groot Bert L.
Published: December 2014

Computational protein design requires methods to accurately estimate free energy changes in protein stability or binding upon an amino acid mutation. From the different approaches available, molecular dynamics‐based alchemical free energy calculations are unique in their accuracy and solid theoretical basis. The challenge in using these methods lies in the need to generate hybrid structures and topologies representing two physical states of a system. A custom made hybrid topology may prove useful for a particular mutation of interest, however, a high throughput mutation analysis calls for a more general approach. In this work, we present an automated procedure to generate hybrid structures and topologies for the amino acid mutations in all commonly used force fields. The described software is compatible with the Gromacs simulation package. The mutation libraries are readily supported for five force fields, namely Amber99SB, Amber99SB*‐ILDN, OPLS‐AA/L, Charmm22*, and Charmm36. © 2014 The Authors Journal of Computational Chemistry Published by Wiley Periodicals, Inc.
Local response dispersion method in periodic systems: Implementation and assessment
Authors: Ikabata Yasuhiro , Tsukamoto Yusuke , Imamura Yutaka , Nakai Hiromi
Published: December 2014

We report the implementation of the local response dispersion (LRD) method in an electronic structure program package aimed at periodic systems and an assessment combined with the Perdew–Burke–Ernzerhof (PBE) functional and its revised version (revPBE). The real‐space numerical integration was implemented and performed exploiting the electron distribution given by the plane‐wave basis set. The dispersion‐corrected density functionals revPBE+LRD was found to be suitable for reproducing energetics, structures, and electron distributions in simple substances, molecular crystals, and physical adsorptions. © 2014 Wiley Periodicals, Inc.
Transcription Factor MEF2C Suppresses Endothelial Cell Inflammation via Regulation of NF‐κB and KLF2
Authors: Xu Zhenhua , Yoshida Takeshi , Wu Lijuan , Maiti Debasish , Cebotaru Liudmila , Duh Elia J.
Published: December 2014

Endothelial cells play a major role in the initiation and perpetuation of the inflammatory process in health and disease, including their pivotal role in leukocyte recruitment. The role of pro‐inflammatory transcription factors in this process has been well‐described, including NF‐κB. However, much less is known regarding transcription factors that play an anti‐inflammatory role in endothelial cells. Myocyte enhancer factor 2 C (MEF2C) is a transcription factor known to regulate angiogenesis in endothelial cells. Here, we report that MEF2C plays a critical function as an inhibitor of endothelial cell inflammation. Tumor necrosis factor (TNF)‐α inhibited MEF2C expression in endothelial cells. Knockdown of MEF2C in endothelial cells resulted in the upregulation of pro‐inflammatory molecules and stimulated leukocyte adhesion to endothelial cells. MEF2C knockdown also resulted in NF‐κB activation in endothelial cells. Conversely, MEF2C overexpression by adenovirus significantly repressed TNF‐α induction of pro‐inflammatory molecules, activation of NF‐κB, and leukocyte adhesion to endothelial cells. This inhibition of leukocyte adhesion by MEF2C was partially mediated by induction of KLF2. In mice, lipopolysaccharide (LPS)‐induced leukocyte adhesion to the retinal vasculature was significantly increased by endothelial cell‐specific ablation of MEF2C. Taken together, these results demonstrate that MEF2C is a novel negative regulator of inflammation in endothelial cells and may represent a therapeutic target for vascular inflammation. J. Cell. Physiol. 230: 1310–1320, 2015. © 2014 Wiley Periodicals, Inc., A Wiley Company
Optimization of bioleaching conditions for metal removal from spent zinc‐manganese batteries using response surface methodology
Authors: Niu Zhirui , Huang Qifei , Xin Baoping , Qi Chenhao , Hu JunFei , Chen Shi , Li Yuping
Published: December 2014

BACKGROUND

As a special residue containing zinc and manganese, spent Zn–Mn batteries cause a serious concern due to their toxicity, abundance and permanence in the environment, and biotechnological recovery of Zn and Mn is one method of recycling this waste. In this study identification of the optimum values of the effective parameters in biotechnological processes was investigated using response surface methodology.

RESULTS

The released dose of Zn and Mn was highly dependent on many parameters. Non‐linear equations were formulated to describe the relationship between bioleaching efficiency of Zn and Mn and four important parameters. The optimum parameter values were determined as follows: dose of mixed energy substrates, exogenous‐acid pH adjustment, incubating temperature and pulp density of 28 g L−1, 1.9, 33 °C, 9.7%, respectively, for Zn release and 29 g L−1, 1.8, 36.7 °C, 8%, respectively, for Mn mobilization. The maximum predictive extraction efficiency of Zn and Mn was 52.5% and 52.4%, respectively.

CONCLUSION

The maximum efficiency of extraction of both Zn and Mn reached 50% under optimum conditions after 9 days of bioleaching. Additionally, XRD analyses suggested that Zn and Mn existed mainly as Hetaerolite (ZnMn2O4) in the spent Zn–Mn batteries, which gradually disappeared and released Zn2+ and Mn2+ into solution during bioleaching process. © 2014 Society of Chemical Industry
Measurement of impulsive choice in rats: Same‐ and alternate‐form test–retest reliability and temporal tracking
Authors: Peterson Jennifer R. , Hill Catherine C. , Kirkpatrick Kimberly
Published: December 2014

Impulsive choice is typically measured by presenting smaller–sooner (SS) versus larger–later (LL) rewards, with biases towards the SS indicating impulsivity. The current study tested rats on different impulsive choice procedures with LL delay manipulations to assess same‐form and alternate‐form test–retest reliability. In the systematic‐GE procedure (Green & Estle, 2003), the LL delay increased after several sessions of training; in the systematic‐ER procedure (Evenden & Ryan, 1996), the delay increased within each session; and in the adjusting‐M procedure (Mazur, 1987), the delay changed after each block of trials within a session based on each rat's choices in the previous block. In addition to measuring choice behavior, we also assessed temporal tracking of the LL delays using the median times of responding during LL trials. The two systematic procedures yielded similar results in both choice and temporal tracking measures following extensive training, whereas the adjusting procedure resulted in relatively more impulsive choices and poorer temporal tracking. Overall, the three procedures produced acceptable same form test–retest reliability over time, but the adjusting procedure did not show significant alternate form test–retest reliability with the other two procedures. The results suggest that systematic procedures may supply better measurements of impulsive choice in rats.
Central hepatectomy with pancreatoduodenectomy for diffusely spread bile duct cancer
Authors: Mizuno Takashi , Kanemoto Hideyuki , Sugiura Teiichi , Okamura Yukiyasu , Uesaka Katsuhiko
Published: December 2014

Background

Major hepatectomy with concomitant pancreatoduodenectomy (major‐HPD) is the only procedure that provides a curative resection for diffusely spread extrahepatic bile duct cancer (DSEBDC). We sometimes encounter patients who cannot undergo major‐HPD because of a poor functional hepatic reserve. The aim of the present study was to assess the feasibility of central hepatectomy with pancreatoduodenectomy (central‐HPD) for patients with DSEBDC as an alternative to major‐HPD.

Methods

Between 2002 and 2010, six patients with DSEBDC underwent central‐HPD. The hepatectomy procedures for central‐HPD included central bisectionectomy with S1 resection (S1r) and right anterior sectionectomy with S1r.

Results

The estimated resection liver volume was decreased from 77.5% to 46.6% by the application of central‐HPD. The median operative duration was 929 min, and the median blood loss was 2568 ml. Postoperative complications were observed in five patients. The proximal ductal stump was histologically positive with non‐invasive cancer in three patients and positive with invasive cancer in one. The overall survival of the six patients was 62.5% at 5 years after surgery.

Conclusions

While central‐HPD was a technically complicated procedure associated with a high morbidity rate, it offered a favorable overall survival and might be an alternative option for DSEBDC patients with a poor hepatic functional reserve.