bluuebunny/crossref_metadata_2025_split huggingface dataset:
Validation of a multiparameter flow cytometry method for the determination of phosphorylated extracellular‐signal‐regulated kinase and DNA in circulating tumor cells
Authors: Pluim Dick , Devriese Lot A. , Beijnen Jos H. , Schellens Jan H. M. 
Published: April 2012

AbstractA simple, selective, and sensitive multiparameter fluorescence activated cell sorting method utilizing density gradient centrifugation and magnetic antibody cell sorting was developed and validated for the determination of phosphorylated extracellular‐signal‐regulated kinase (pERK) and DNA in circulating tumor cells (CTCs). Cell preparation tubes (CPT) were used for peripheral blood collection and density gradient centrifugation, followed by phosphorylation of ERK with epidermal growth factor (EGF). After fixation with formaldehyde and methanol, magnetic anti epithelial cell adhesion molecule (EpCAM) micro‐beads were used for the selective isolation of CTCs from the background, consisting of peripheral blood mononuclear cells and platelets. Subsequently, samples were stained with Hoechst 33342, and fluorescent antibodies against EpCAM, CD45, and pERK. Flow cytometry was used for identification and enumeration of CTCs and determination of their pERK and DNA content. The validation parameters included specificity, recovery, linearity, precision, sensitivity, and stability. The lower limit of quantification was two CTCs per 8 ml peripheral blood. Samples were stable for 4 months in storage at −80°C. The applicability of the method was demonstrated by successful enumeration of CTCs, and the determination of DNA, and pERK before and after stimulation with EGF in 8 ml peripheral blood samples from patients with metastatic cancer. © 2012 International Society for Advancement of Cytometry
A novel multiparameter flow cytometric assay for inosine triphosphatase expression analysis in leukocytes
Authors: Vroemen Wim H. M. , Munnix Imke C. A. , Bakker Jaap A. , Bierau Jörgen , Huts Mark , Leers Mathie P. G. 
Published: April 2012

AbstractThe aim of this study was to assess inosine triphosphate (ITPase) expression in the different leukocyte populations present in peripheral blood samples of a nonimmune compromised control group. For this purpose, a multiparameter flow cytometric assay was developed and performed to study ITPase expression in peripheral leukocyte subpopulations of healthy volunteers (n = 20). Qualitative ITPase expression was assessed by determining the percentage of ITPase‐positive cells. Quantitative data were obtained by measuring the median fluorescent intensity (MFI). Subcellular localization of ITPase was analyzed using immunocytochemistry. Immunocytochemistry showed that ITPase is present in all leukocytes and localized intracellular. Based on this finding, a multiparameter flow cytometric assay was developed using a Fix & Perm strategy. Qualitative and quantitative ITPase expression remained stable (variation, <10%) for at least 48 h after blood sampling. MFI values showed that activated monocytes contained significantly more ITPase when compared to the total monocyte fraction (P < 0.0001), which subsequently had a higher amount of expression than granulocytes (P < 0.0001). In addition, the phagocyte subpopulations ([activated] monocytes and granulocytes) contained significantly higher levels of ITPase when compared to lymphocytes (P < 0.0001). Within the lymphocyte fraction, it appeared that T‐helper cells contained significantly higher ITPase levels when compared to cytotoxic T cells, B lymphocytes, and natural killer cells (P < 0.0001). Our study is the first which describes a flow cytometry assay to analyze ITPase expression in leukocytes qualitatively as well as quantitatively and visualizes the intracellular localization of ITPase in leukocytes. © 2012 International Society for Advancement of Cytometry
Practical guidelines for the high‐sensitivity detection and monitoring of paroxysmal nocturnal hemoglobinuria clones by flow cytometry
Authors: Sutherland D. Robert , Keeney Michael , Illingworth Andrea 
Published: April 2012

AbstractBackground:Paroxysmal nocturnal hemoglobinuria (PNH) is a life‐threatening disorder caused by an inability to make glyco‐phosphatidyl‐inositol (GPI) anchors. While flow cytometry is the method of choice to detect the loss of GPI‐linked proteins, the development and validation of sensitive, standardized, methodologies have been hampered by the rarity of this disease and by technical difficulties in the accurate identification of PNH cells.Methods:Guidelines for the diagnosis and monitoring of PNH by flow cytometry were recently published by the International Clinical Cytometry Society (ICCS). However, specific reagent cocktails, and associated detailed analytic strategies were not directly addressed therein. In this supporting document based on the ICCS guidelines, we provide concise practical protocols for the high‐sensitivity detection of PNH RBCs and WBCs (both granulocytes and monocytes).Results:The CD235aFITC/CD59PE assay described was capable of detecting as few as 20 Type III PNH RBCs per million cells. Frequencies of Type III PNH cells in 10 normal samples were in the 0–6 per million RBCs. The high‐resolution granulocyte/neutrophil assays described in this study could detect PNH phenotypes consistently at a level of 0.01% sensitivity. Frequencies of PNH phenotypes in normal individuals were in the 0–10 per million granulocytes/neutrophils range.Conclusions:The careful screening and selection of specific antibody conjugates has allowed the development of reagent cocktails suitable for high‐sensitivity flow cytometric detection of PNH RBCs and PNH WBCs. The reagent cocktails described herein can be used on a variety of clinical flow cytometers equipped with four or more photo multiplier tubes. © 2012 International Clinical Cytometry Society
A roadside unit‐based localization scheme for vehicular ad hoc networks
Authors: Ou Chia‐Ho 
Published: April 2012

SUMMARYVehicular Ad Hoc Networks (VANETs), designed to ensure the safety and comfort of passengers via the exchange of information amongst nearby vehicles or between the vehicles and Roadside Units (RSUs), have attracted particular attention. However, the success of many VANET applications depends on their ability to estimate the vehicle position with a high degree of precision, and thus, many vehicle localization schemes have been proposed. Many of these schemes are based on vehicle‐mounted Global Positioning System (GPS) receivers. However, the GPS signals are easily disturbed or obstructed. Although this problem can be resolved by vehicle‐to‐vehicle communication schemes, such schemes are effective only in VANETs with a high traffic density. Accordingly, this paper presents a VANET localization scheme in which each vehicle estimates its location on the basis of beacon messages broadcast periodically by pairs of RSUs deployed on either side of the road. In addition, three enhancements to the proposed scheme are presented for the RSU deployment, RSU beacon collisions, and RSU failures. Overall, the ns‐2 simulation results show that the localization scheme achieves a lower localization error than existing solutions on the basis of vehicle‐to‐vehicle communications and is robust toward changes in the traffic density and the vehicle speed. Copyright © 2012 John Wiley & Sons, Ltd.
A Gal4/UAS system for conditional transgene expression in rhombomere 4 of the zebrafish hindbrain
Authors: Choe Seong‐Kyu , Nakamura Mako , Ladam Franck , Etheridge Letitiah , Sagerström Charles G. 
Published: April 2012

AbstractBackground: The zebrafish is well established as a model organism for the study of vertebrate embryogenesis, but transgenic lines enabling restricted gene expression are still lacking for many tissues. Results: We first generated the hoxb1a(β−globin):eGFPum8 line that expresses eGFP in hindbrain rhombomere 4 (r4), as well as in facial motor neurons migrating caudally from r4. Second, we generated the hoxb1a(β‐globin) Gal4VP16um60 line to express the exogenous Gal4VP16 transcription factor in r4. Lastly, we prepared the UAS(β‐actin):hoxa3aum61 line where the hoxa3a gene, which is normally expressed in r5 and r6, is under control of Gal4‐regulated UAS elements. Crossing the hoxb1a(β−globin):Gal4VP16um60 line to the UAS(β‐actin):hoxa3aum61 line drives robust hoxa3a expression in r4. We find that transgenic expression of hoxa3a in r4 does not affect hoxb1a expression, but has variable effects on migration of facial motorneurons and formation of Mauthner neurons. While cases of somatic transgene silencing have been reported in zebrafish, we have not observed such silencing to date, possibly because of our efforts to minimize repetitive sequences in the transgenic constructs. Conclusion: We have generated three transgenic lines that will be useful for future studies by permitting the labeling of r4‐derived cells, as well as by enabling r4‐specific expression of various transgenes. Developmental Dynamics 241:1125–1132, 2012. © 2012 Wiley Periodicals, Inc.
NaEu3(GeO4)2(OH)2: A High‐Pressure‐Stable Photoluminescent Lanthanide Germanate
Authors: Zhao Xinguang , Yan Tingting , Wang Kai , Yan Yan , Zou Bo , Yu Jihong , Xu Ruren 
Published: April 2012

AbstractThe europium germanate NaEu3(GeO4)2(OH)2 was prepared under mild hydrothermal conditions. In situ synchrotron X‐ray diffraction analysis and photoluminescence spectroscopic measurement in a diamond anvil cell (DAC) under high pressure were performed to study its structural stability as well as the luminescent properties of the Eu3+ ions. The synchrotron X‐ray diffraction analysis revealed that the structure of NaEu3(GeO4)2(OH)2 was stable under pressure that ranged from 1 atm to 20 GPa with only a little distortion, and the unit‐cell compression occurred more readily along the a axis than along the c axis. When the pressure was reduced to ambient pressure, the distortion and the unit cell recovered to the original state. Photoluminescence spectroscopic studies under high pressure showed that the redshift of the 5D0 → 7FJ (J = 0, 1, 2) transitions and the pressure imposed on the sample had a good linear relationship. Also, the intensity of the red emission lines was independent of the pressure in the range from 9 to about 20 GPa.
Subspace system identification of support‐excited structures—part I: theory and black‐box system identification
Authors: Kim Junhee , Lynch Jerome P. 
Published: April 2012

SUMMARYThis paper reviews the theoretical principles of subspace system identification as applied to the problem of estimating black‐box state‐space models of support‐excited structures (e.g., structures exposed to earthquakes). The work distinguishes itself from past studies by providing readers with a powerful geometric interpretation of subspace operations that relates directly to theoretical structural dynamics. To validate the performance of subspace system identification, a series of experiments are conducted on a multistory steel frame structure exposed to moderate seismic ground motions; structural response data is used off‐line to estimate black‐box state‐space models. Ground motions and structural response measurements are used by the subspace system identification method to derive a complete input–output state‐space model of the steel frame system. The modal parameters of the structure are extracted from the estimated input–output state‐space model. With the use of only structural response data, output‐only state‐space models of the system are also estimated by subspace system identification. The paper concludes with a comparison study of the modal parameters extracted from the input–output and output‐only state‐space models in order to quantify the uncertainties present in modal parameters extracted from output‐only models. Copyright © 2012 John Wiley & Sons, Ltd.
Psychopathological and Clinical Features of Remitted Anorexia Nervosa Patients: A Six‐year Follow‐up Study
Authors: Lo Sauro Carolina , Castellini Giovanni , Lelli Lorenzo , Faravelli Carlo , Ricca Valdo 
Published: April 2012

AbstractObjectiveRemission from anorexia nervosa (AN) is a controversial issue, as remitted individuals have been found to show residual anorectic attitudes and concerns about weight and shape. The aims of this study were to evaluate the psychopathological features of remitted AN subjects 6 years after the end of a cognitive behavioural therapy and the predictors of reduction in psychopathology.MethodsThe sample was composed of 134 AN subjects, evaluated at baseline, at the end of treatment, 3 and 6 years after the end of treatment, by means of the Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition and several self‐reported questionnaires.ResultsRemitted patients at 6 years of follow‐up still showed higher eating and shape concerns, compared with healthy controls. Duration of illness, obsessive‐compulsive and depressive symptoms were moderators of change in psychopathology across time.DiscussionPsychopathological features represent an enduring trait for AN patients. General psychopathology showed different effects on symptoms reduction across time. Copyright © 2012 John Wiley & Sons, Ltd and Eating Disorders Association.
The Science Behind Pre‐Columbian Evidence of Syphilis in Europe: Research by Documentary
Authors: Armelagos George J. , Zuckerman Molly K. , Harper Kristin N. 
Published: April 2012

AbstractThis article discusses the presentation of scientific findings by documentary, without the process of peer review. We use, as an example, PBS's “The Syphilis Enigma,” in which researchers presented novel evidence concerning the origin of syphilis that had never been reviewed by other scientists. These “findings” then entered the world of peer‐reviewed literature through citations of the documentary itself or material associated with it. Here, we demonstrate that the case for pre‐Columbian syphilis in Europe that was made in the documentary does not withstand scientific scrutiny. We also situate this example from paleopathology within a larger trend of “science by documentary” or “science by press conference,” in which researchers seek to bypass the peer review process by presenting unvetted findings directly to the public. © 2012 Wiley Periodicals, Inc.
The Role of Terrestriality in Promoting Primate Technology
Authors: Meulman Ellen J.M. , Sanz Crickette M. , Visalberghi Elisabetta , van Schaik Carel P. 
Published: April 2012

Abstract“Complex technology” has often been considered a hallmark of human evolution. However, recent findings show that wild monkeys are also capable of habitual tool use. Here we suggest that terrestriality may have been of crucial importance for the innovation, acquisition, and maintenance of “complex” technological skills in primates. Here we define complex technological skills as tool‐use variants that include at least two tool elements (for example, hammer and anvil), flexibility in manufacture or use (that is, tool properties are adjusted to the task at hand), and that skills are acquired in part by social learning. Four lines of evidence provide support for the terrestriality effect. First, the only monkey populations exhibiting habitual tool use seem to be particularly terrestrial. Second, semi‐terrestrial chimpanzees have more complex tool variants in their repertoire than does their arboreal Asian relative, the orangutan. Third, tool variants of chimpanzees used in a terrestrial setting tend to be more complex than those used exclusively in arboreal contexts. Fourth, the higher frequency in tool use among captive versus wild primates of the same species may be attributed in part to a terrestriality effect. We conclude that whereas extractive foraging, intelligence, and social tolerance are necessary for the emergence of habitual tool use, terrestriality seems to be crucial for acquiring and maintaining complex tool variants, particularly expressions of cumulative technology, within a population. Hence, comparative evidence among primates supports the hypothesis that the terrestriality premium may have been a major pacemaker of hominin technological evolution. © 2012 Wiley Periodicals, Inc.
Cytotoxicity and cell imaging potentials of submicron color‐tunable yttria particles
Authors: Atabaev Timur Sh. , Lee Jong Ho , Han Dong‐Wook , Hwang Yoon‐Hwae , Kim Hyung‐Kook 
Published: April 2012

AbstractIncreased demand of environment protection encouraged scientists to design products and processes that minimize the use and generation of hazardous substances. This work presents comprehensive result of large‐scale fabrication and investigation of red‐to‐green tunable submicron spherical yttria particles codoped with low concentrations of Eu+3 and Tb+3. The color emission of synthesized particles can be precisely tuned from red to green by simple variation of Tb/Eu ratio and excitation wavelength. The Tb/Eu‐codoped Y2O3 particles did not adversely affect the viability of L‐929 fibroblastic cells at concentrations less than 62.5 ppm. Through internalization and wide distribution inside the cells, Tb/Eu codoped Y2O3 particles with intense bright green or red fluorescence rendered cell imaging to be possible. The high brightness, excellent stability, low‐toxicity, and imaging capability along with fine color‐tunability of synthesized particles enable to find promising application in various areas. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 100A: 2287–2294, 2012.
The role of macrocyclic ligands in the peroxo/superoxo nature of Ni–O2 biomimetic complexes
Authors: Zapata‐Rivera Jhon , Caballol Rosa , Calzado Carmen J. 
Published: April 2012

AbstractThe impact of the macrocyclic ligand on the electronic structure of two LNiO2 biomimetic adducts, [Ni(12‐TMC)O2]+ (12‐TMC = 1,4,7,10‐tetramethyl‐1,4,7,10‐tetraazacyclododecane) and [Ni(14‐TMC)O2]+ (14‐TMC = 1,4,8,11‐tetramethyl‐1,4,8,11‐tetraazacyclotetradecane), has been inspected by means of difference‐dedicated configuration interaction calculations and a valence bond reading of the wavefunction. The system containing the 12‐membered macrocyclic ligand has been experimentally described as a side‐on nickel(III)‐peroxo complex, whereas the 14‐membered one has been characterized as an end‐on nickel(II)‐superoxide. Our results put in evidence the relationship between the steric effect of the macrocyclic ligand, the O2 coordination mode and the charge transfer extent between the Ni center and the O2 molecule. The 12‐membered macrocyclic ligand favors a side‐on coordination, a most efficient overlap between Ni 3d and O2 π* orbitals and, consequently, a larger charge transfer from LNi fragment to O2 molecule. The analysis of the ground‐state electronic structure shows an enhancement of the peroxide nature of the NiO2 interaction for [Ni(12‐TMC)O2]+, although a dominant superoxide character is found for both systems. © 2012 Wiley Periodicals, Inc.
DCMB that combines divide‐and‐conquer and mixed‐basis set methods for accurate geometry optimizations, total energies, and vibrational frequencies of large molecules
Authors: Wu Anan , Xu Xin 
Published: April 2012

AbstractWe present a method, named DCMB, for the calculations of large molecules. It is a combination of a parallel divide‐and‐conquer (DC) method and a mixed‐basis (MB) set scheme. In this approach, atomic forces, total energy and vibrational frequencies are obtained from a series of MB calculations, which are derived from the target system utilizing the DC concept. Unlike the fragmentation based methods, all DCMB calculations are performed over the whole target system and no artificial caps are introduced so that it is particularly useful for charged and/or delocalized systems. By comparing the DCMB results with those from the conventional method, we demonstrate that DCMB is capable of providing accurate prediction of molecular geometries, total energies, and vibrational frequencies of molecules of general interest. We also demonstrate that the high efficiency of the parallel DCMB code holds the promise for a routine geometry optimization of large complex systems. © 2012 Wiley Periodicals, Inc.
Imaging of prion diseases
Authors: Letourneau‐Guillon Laurent , Wada Ryan , Kucharczyk Walter 
Published: April 2012

AbstractPrion diseases are caused by self‐replicating proteins that induce lethal neurodegenerative disorders. In the last decade, the understanding of the different clinical, pathological, and neuroimaging phenotypes of this group of disorders has evolved paralleling the advances in prion molecular biology. From an imaging standpoint, the implementation of diffusion‐weighted imaging in routine practice has markedly facilitated the detection of prion diseases, especially Creutzfeldt‐Jakob. Less frequent prion‐related disorders, including genetic diseases, may also benefit from progresses in the field of quantitative diffusion‐weighted imaging, MR spectroscopy or molecular imaging. Herein, we present a review of the neuroimaging features of the prion disorders known to affect humans emphasizing the important contribution of MRI in the diagnosis of this group of disorders. J. Magn. Reson. Imaging 2012;35:998‐1012. © 2012 Wiley Periodicals, Inc.
Genotyping of 11 human papillomaviruses by multiplex PCR with a GeXP analyzer
Authors: Yang Meng‐Jie , Luo Le , Nie Kai , Wang Miao , Zhang Chen , Li Jin , Ma Xue‐jun 
Published: April 2012

AbstractA new, rapid, and high‐throughput method was developed for simultaneous detection of 11 human papillomavirus (HPV) genotypes including nine high‐risk types (HPV16, 18, 31, 33, 35, 39, 52, 58, and 66) and two low‐risk types (HPV6 and 11) in a single tube by multiplex PCR based on a GenomeLab Gene Expression Profiler (GeXP) analyzer (GeXP‐PCR). Eleven sets of chimeric primers were used to initiate the PCR, and one pair of universal primers was used for the subsequent cycles of the PCR. The specificity of GeXP‐PCR for each HPV type was examined with clinical samples of single type HPV infection tested previously. The sensitivity of GeXP‐PCR was evaluated by performing the assay on serial 10‐fold dilutions of cloned PCR products. The GeXP‐PCR achieved a sensitivity of 100 copies when all of the 11 pre‐mixed plasmids containing HPV targets were present. Analyses of 124 clinical specimens using the GeXP‐PCR demonstrated that the GeXP‐PCR assay had comparable sensitivity and specificity to those of reported multiple PCR assay and an increased detection of HPV 11 in samples with mixed infections. In conclusion, the GeXP‐PCR is a fast, sensitive, and high throughput method for the detection of multiple HPV infections. J. Med. Virol. 84:957–963, 2012. © 2012 Wiley Periodicals, Inc.
Hepatitis B virus X protein blocks filamentous actin bundles by interaction with eukaryotic translation elongat ion factor 1 alpha 1
Authors: Lin Wan‐Song , Jiao Bo‐Yan , Wu Yun‐Li , Chen Wan‐Nan , Lin Xu 
Published: April 2012

AbstractHepatitis B virus (HBV)‐encoded X protein (HBx protein) is a multi‐functional regulatory protein. It functions by protein–protein interaction and plays a pivotal role in the pathogenesis of HBV‐related diseases. However, the partners in hepatocytes interacting with HBx protein are far from understood fully. In this study, immunoprecipitation was employed to screen for binding partners for the HBx protein from huh‐7 hepatoma cells infected with recombinant adenovirus expressing HBx protein, and five cellular proteins including eukaryotic translation elongation factor 1 alpha 1 (eEF1A1), were identified. The interaction between HBx protein and eEF1A1 was confirmed further using a GST pull‐down assay and co‐immunoprecipitation, respectively. In Huh‐7 hepatoma cells, the HBx protein inhibits dimer formation of eEF1A1, hence blocks filamentous actin bundling. These findings provide new insights into the molecular mechanisms involved in the functions of the HBx protein. J. Med. Virol. 84:871–877, 2012. © 2012 Wiley Periodicals, Inc.