bluuebunny/crossref_metadata_2025 huggingface dataset:
Automated sample area definition for high‐throughput microscopy
Authors: Zeder M. , Ellrott A. , Amann R.
Published: March 2011

High‐throughput screening platforms based on epifluorescence microscopy are powerful tools in a variety of scientific fields. Although some applications are based on imaging geometrically defined samples such as microtiter plates, multiwell slides, or spotted gene arrays, others need to cope with inhomogeneously located samples on glass slides. The analysis of microbial communities in aquatic systems by sample filtration on membrane filters followed by multiple fluorescent staining, or the investigation of tissue sections are examples. Therefore, we developed a strategy for flexible and fast definition of sample locations by the acquisition of whole slide overview images and automated sample recognition by image analysis. Our approach was tested on different microscopes and the computer programs are freely available (http://www.technobiology.ch). © 2011 International Society for Advancement of Cytometry
A novel flow cytometry‐based assay to study leukocyte–endothelial cell interactions in vitro
Authors: Kränkel Nicolle , Kuschnerus Kira , Madeddu Paolo , Lüscher Thomas F. , Landmesser Ulf
Published: March 2011

Interactions between peripheral blood mononuclear cells (PBMC) and the endothelium critically determine vascular repair and reparative angiogenesis, but also pathological processes, such as atherosclerosis. Current methodology to study these interactions mostly regards PBMC as a homogenous population or it restricts its focus on individual cell types and therefore does not appreciate the differential behavior of individual PBMC subpopulations, which synergize or antagonize each other to obtain the overall effect. We therefore developed a flow cytometry‐based in vitro assay to assess multiple parameters of interaction between several individual populations of PBMC and an endothelial monolayer. Freshly isolated, unlabelled human PBMC were left to adhere to or transmigrate through a monolayer of fluorescence‐labeled human aortic endothelial cells grown to confluence on the filter membrane of sterile transwell migration inserts. Monocyte chemoattractant protein‐1 (MCP‐1) was applied as a chemoattractant to the lower compartment of the migration chamber. After 6 h, transmigrating PBMC were harvested from the lower compartment, while nonadherent and adhering cell populations were harvested from the upper compartment by sequential washing/detachment. All three cell fractions were then individually stained with fluorescence‐labeled monoclonal antibodies and analyzed by flow cytometry. Quantification was achieved by the usage of counting beads. Endothelial cells were separated from PBMC during the analysis by a multiparametric gating strategy. Using the newly established assay, we observed distinct migration patterns for inflammatory CD14hiCD16neg and resident CD16pos monocytes. These cell types differed in their basal adhesion and transmigration patterns as well as their responses to the CCR2 ligand MCP‐1. This assay allows for the parallel study of interactions between multiple individual leukocyte populations and an endothelial layer. Several readouts can be derived from the same experiment, like the composition of adhering and transmigrating cell fractions or the individual adhesion/migration behavior of several distinct cell types. © 2011 International Society for Advancement of Cytometry
Improved ZAP‐70 assay using two clones, multiple methods of analysis and clinical correlation
Authors: Degheidy Heba A. , Venzon David J. , Farooqui Mohammed Z. H. , Abbasi Fatima , Arthur Diane C. , Wilson Wyndham H. , Wiestner Adrian , Stetler‐Stevenson M. A. , Marti Gerald E.
Published: March 2011

Introduction:

In a companion methodological study, we compared two anti‐ZAP‐70 clones (1E7.2 AF 488 and SBZAP PE) and four selected methods of analysis. Clinical correlations are required for validation.

Methods:

Multicolor flow‐cytometric evaluation of ZAP‐70, CD38, CD69, CD26, CD49d, and CD27 was tested in 45 untreated‐CLL patients. Four methods of ZAP‐70 expression analysis and a scoring system were designed. A correlation analysis between ZAP‐70 score, immunoglobulin heavy chain variable (IGHV) mutational status, fluorescence in situ hybridization, and these biomarkers was undertaken.

Results:

There is a strong correlation between ZAP‐70 expression and IGHV mutational status. The scoring system for a single reagent (P = 0.0006 or 0.0002) favors the use of multiple methods of analysis. The combined score was substantially equivalent (P = 0.0003). There was also a correlation with del 13q14 (P = 0.017) and trisomy12 (P = 0.011). A correlation for CD38 and ZAP‐70 score was seen using both 1E7.2 AF488 and SBZAP PE when ≥20% or ≥7% cutoff was used. A positive correlation was seen for CD49d expression using both reagents. CD26 showed a correlation with ZAP‐70 expression, but it was dependent upon the method of analysis. CD69 and CD27 showed no statistically significant correlation.

Conclusion:

In our study population, ZAP‐70 expression is the better predictor of the IGHV mutational status. The correlation analysis confirms that the use of four methods of analysis with a single reagent or both reagents is superior to the use of a single method of analysis. The routine use of CD38, CD49d, and CD26 will require standardization. Published 2011 Wiley‐Liss, Inc.
Identity of the avian wing digits: Problems resolved and unsolved
Authors: Young Rebecca L. , Bever Gabe S. , Wang Zhe , Wagner Günter P.
Published: March 2011

Controversy over bird wing digit identity has been a touchstone for various ideas in the phylogeny of birds, homology, and developmental evolution. This review summarizes the past 10 years of progress toward understanding avian digit identity. We conclude that the sum of evidence supports the Frame Shift Hypothesis, indicating that the avian wing digits have changed anatomical location. Briefly, the derivation of birds from theropod dinosaurs and the positional identities of the avian wing digits as 2, 3, and 41 are no longer in question. Additionally, increasing evidence indicates that the developmental programs for identity of the wing digits are of digits I, II, and III. Therefore, the attention moves from whether the digit identity frame shift occurred, to what the mechanisms of the frame shift were, and when in evolution it happened. There is considerable uncertainty about these issues and we identify exciting new research directions to resolve them. Developmental Dynamics 240:1042–1053, 2011. © 2011 Wiley‐Liss, Inc.
Exogenous fibroblast growth factor 8 rescues development of mouse diastemal vestigial tooth ex vivo
Authors: Li Lu , Yuan Guohua , Liu Chao , Zhang Lu , Zhang Yanding , Chen YiPing , Chen Zhi
Published: March 2011

Regression of vestigial tooth buds results in the formation of the toothless diastema, a unique feature of the mouse dentition. Revitalization of the diastemal vestigial tooth bud provides an excellent model for studying tooth regeneration and replacement. It has been previously shown that suppression of fibroblast growth factor (FGF) signaling in the diastema results in vestigial tooth bud regression. In this study, we report that application of exogenous FGF8 to the mouse embryonic diastemal region rescues diastemal tooth development. However, this rescue of diastemal tooth development occurs only in an isolated diastemal regions and not in the mandibular quadrant, which includes the incisor and molar germs. FGF8 promotes cell proliferation and inhibits apoptosis in diastemal tooth epithelium, and revitalizes the tooth developmental program, as evidenced by the expression of genes critical for normal tooth development. Our results also support the idea that the adjacent tooth germs contribute to the suppression of diastemal vestigial tooth buds by means of multiple signals. Developmental Dynamics 240:1344–1353, 2011. © 2011 Wiley‐Liss, Inc.
The upside of an annual survey in light of involvement and use: Evaluating the Advanced Technological Education program
Authors: Toal Stacie A. , Gullickson Arlen R.
Published: March 2011

In 1999, the National Science Foundation (NSF) awarded funds to the Evaluation Center at Western Michigan University to conduct an external evaluation of the Advanced Technological Education (ATE) program. ATE, a federally mandated program designed to increase the number and quality of skilled technicians in the U.S. workforce, has funded over 346 projects and centers across the nation. This case study describes the relationship between project‐level involvement in the ATE program evaluation and the use and influence of the evaluation on project primary investigators and evaluators. Although this large, multisite program evaluation employed numerous evaluative data‐collection and dissemination techniques, project leaders and evaluators associated the program evaluation primarily with an annual Web‐based survey. The NSF's expectation that projects would complete the annual survey contributed to feelings of involvement and, in many cases, promoted use and impact. © Wiley Periodicals, Inc., and the American Evaluation Association.
Documenting the impact of multisite evaluations on the science, technology, engineering, and mathematics field
Authors: Roseland Denise , Greenseid Lija O. , Volkov Boris B. , Lawrenz Frances
Published: March 2011

This chapter discusses the impact that four multisite National Science Foundation (NSF) evaluations had on the broader field of science, technology, engineering, and mathematics education and evaluation. Three approaches were used to investigate the broader impact of these evaluations on the field: (a) a citation analysis, (b) an on‐line survey, and (c) surveys and follow‐up interviews with NSF project principal investigators (PIs). The results indicate that the four evaluations had at least some impact. In addition, they show that the four efforts had differential impact, generally related to the way each evaluation operated. Evaluations in operation for longer time periods produced more products with more types of dissemination. The most‐cited products tended to be instruments and tools. The field influence survey showed that, overall, few NSF PIs had any knowledge of influential evaluations, evaluation publications, or evaluators. © Wiley Periodicals, Inc., and the American Evaluation Association.
Reflection on four multisite evaluation case studies
Authors: Brandon Paul R.
Published: March 2011

What do the findings of four National Science Foundation evaluation case studies suggest to an evaluation scholar who was not part of the research team that created them? This chapter carefully reviews the cases and summarizes their comparative findings. The four Beyond Use case studies add to the literature on levels of evaluation use, with the findings suggesting a level of involvement not surprising for multisite evaluations like these. Other results indicate that the levels of findings use were greater than the levels of stakeholder involvement reported and that a substantial portion of the use reported in the case studies was process use. Interpreting the reported relationships between stakeholder involvement and use must be done cautiously, but it remains true that showing a causal relationship between stakeholder participation in evaluation activities and the use of evaluation findings is as difficult in these case studies as it has been in most of the research on that relationship over the past 30 years. The chapter raises implications for both practice and research on multisite evaluations. © Wiley Periodicals Inc., and the American Evaluation Association.
Toward better research on—and thinking about—evaluation influence, especially in multisite evaluations
Authors: Mark Melvin M.
Published: March 2011

Evaluation is typically carried out with the intention of making a difference in the understandings and actions of stakeholders and decision makers. The author provides a general review of the concepts of evaluation “use,” evaluation “influence,” and “influence pathways,” with connections to multisite evaluations. The study of evaluation influence and influence pathways is briefly discussed. © Wiley Periodicals, Inc., and the American Evaluation Association.
A BACH2‐BCL2L1 fusion gene resulting from a t(6;20)(q15;q11.2) chromosomal translocation in the lymphoma cell line BLUE‐1
Authors: Türkmen Seval , Riehn Mathias , Klopocki Eva , Molkentin Mara , Reinhardt Richard , Burmeister Thomas
Published: March 2011

Abnormalities of the long arm of chromosome 6 are a common feature in various B‐cell malignancies. In most cases, the genes involved have not yet been clearly identified. We have molecularly characterized the recently established Burkitt lymphoma cell line BLUE‐1 that carries a t(6;20)(q15;q11.2) rearrangement in addition to the typical t(8;14) with MYC‐IGH fusion. To identify the gene loci involved on both chromosomes we applied a sequential BAC clone mapping strategy. By using RT‐PCR we were finally able to detect a chimeric mRNA transcript showing a fusion of the first (non‐coding) exon of BACH2 (BTB and CNC homology 1, basic leucine zipper transcription factor 2) on 6q15 to the second exon of BCL2L1 (BCL‐X) on 20q11. Various fusion transcripts were detected for different BCL2L1 (BCL‐XL) isoforms. The fusion ultimately results in strong expression of the BCL2L1 (BCL‐XL) anti‐apoptosis protein, as demonstrated by immunoblotting. This is the first report that shows the involvement of both BCL2L1 and the transcription factor BACH2 in a chromosomal rearrangement. It points to BACH2 as a possibly important target in lymphomas with 6q aberrations, although other genes on 6q are probably also involved in these cases. Moreover, it suggests that members of the BCL2 anti‐apoptosis gene family other than BCL2 itself might also be involved in lymphoma. © 2011 Wiley‐Liss, Inc.
Genomic aberrations in diffuse low‐grade gliomas
Authors: Dahlback Hanne‐Sofie S. , Gorunova Ludmila , Brandal Petter , Scheie David , Helseth Eirik , Meling Torstein R. , Heim Sverre
Published: March 2011

The current classification of diffuse low‐grade gliomas is based mainly on histopathological criteria, which cannot accurately predict the highly variable clinical course observed in patients with such tumors. In an attempt to increase pathogenetic understanding of these tumors, we investigated 38 WHO Grade II astrocytomas, oligodendrogliomas, and oligoastrocytomas using a combination of G‐band chromosome analysis and high‐resolution comparative genomic hybridization (HR‐CGH). Abnormal karyotypes were found in 41% of tumors. Karyotypes of astrocytomas and oligodendrogliomas were near‐diploid whereas oligoastrocytomas also displayed near‐tetraploid clones. The most common aberrations were losses of chromosomes X, Y, 3, 4, 6, and 11 and gains of chromosomes 8 and 12. The only recurrent structural rearrangement was del(6)(q21). HR‐CGH analysis verified karyotyping findings but also revealed frequent losses at 1p, 17q, and 19q and gains of 7q, 10p, 11q, and 20p. Among the tumors were two gemistocytic astrocytomas, a subgroup of diffuse astrocytomas with a particular predisposition for progression but not studied cytogenetically before; one showed a near‐diploid, complex karyotype with structural aberrations of chromosomes 1, 3, and 11 whereas both displayed simple aberrations including loss of 11p by HR‐CGH. Our findings suggest that within diffuse low‐grade gliomas are genetically distinct entities that do not fit the currently used classification. In addition, tumors with complex chromosomal aberrations had a higher tendency for aggressive tumor behavior (shorter progression‐free survival) than tumors displaying simple aberrations only (P= 0.07). This could help identify genetic subsets of patients with low‐grade glioma who might benefit from early antineoplastic therapy. © 2011 Wiley‐Liss, Inc.
Downregulation of methylthioadenosin phosphorylase by homozygous deletion in gastric carcinoma
Authors: Kim Jin , Kim Min A , Min Sun Young , Jee Chang Do , Lee Hee Eun , Kim Woo Ho
Published: March 2011

The methylthioadenosine phosphorylase (MTAP) gene is located on 9p21 telomeric to the CDKN2A tumor suppressor gene. Loss of MTAP gene is frequently associated with CDKN2A homozygous deletion. Although the homozygous deletion of MTAP has been reported in various human cancers, its function in gastric carcinogenesis is unknown. Here, we determined the status of the MTAP gene by using a combination of array‐based comparative genomic hybridization and oligonucleotide microarray. It was found that MTAP was deleted and downregulated in 2 of 10 gastric cancer cell lines. Of the 494 primary gastric carcinomas examined, MTAP expression at the protein level was reduced in 59 (11.9%). Furthermore, a lack of MTAP expression was found to be associated with poor survival (P = 0.038). The genomic loss of MTAP and CDKN2A in gastric carcinomas was investigated by quantitative real‐time PCR. Among 20 gastric carcinomas, two cases showed deletion of both MTAP and CDKN2A, and three samples showed homozygous deletion of MTAP, but not of CDKN2A. An analysis of gastric carcinomas revealed that reduced MTAP expression correlated significantly with a genomic deletion. Furthermore, functional assays by transfecting the siRNA or the expressional cDNA into gastric cancer cell lines demonstrated that MTAP regulates cell growth and invasion. The present study suggests that MTAP plays an important role in the regulation of gastric carcinogenesis and, in particular, that MTAP loss is implicated in some way with tumor growth via the modulation of cellular properties, which, in turn, suggests that MTAP has therapeutic applications. © 2011 Wiley‐Liss, Inc.
Scale invariance and self‐similarity in kinematic wave overland flow in space and time
Authors: Haltas I. , Kavvas M. L.
Published: March 2011

Fractals are famous for their self‐similar nature at different spatial scales. Similar to fractals, solutions of scale invariant processes are self‐similar at different space–time scales. This unique property of scale‐invariant processes can be utilized to translate the solution of the processes at a much larger or smaller space–time scale (domain) based on the solution calculated on the original space–time scale. This study investigates scale invariance conditions of kinematic wave overland flow process in one‐parameter Lie group of point transformations framework. Scaling (stretching) transformation is one of the one‐parameter Lie group of point transformations and it has a unique importance among the other transformations, as it leads to the scale invariance or scale dependence of a process. Scale invariance of a process yields a self‐similar solution at different space–time scales. However, the conditions for the process to be scale invariant usually dictate various relationships between the scaling coefficients of the dependent and independent variables of the process. Therefore, the scale invariance of a process does not assure a self‐similar solution at any arbitrary space and time scale. The kinematic wave overland flow process is modelled mathematically as initial‐boundary value problem. The conditions to be satisfied by the system of governing equations as well as the initial and boundary conditions of the kinematic wave overland flow process are established in order for the process to be scale invariant. Also, self‐similarity of the solution of the kinematic wave overland flow under the established invariance conditions is demonstrated by various numerical example problems. Copyright © 2011 John Wiley & Sons, Ltd.
Toxicity studies of nonylphenol and octylphenol: hormonal, hematological and biochemical effects inClarias gariepinus
Authors: Senthil Kumaran Satyanarayanan , Kavitha Chokkalingam , Ramesh Mathan , Grummt Tamara
Published: March 2011

Among the numerous chemicals discharged into aquatic ecosystems, nonylphenol (NP) and octylphenol (OP) have been shown to have a potent effect on the endocrine system of fish; this issue has been clearly dealt with in several studies. The objective of this study was to assess and compare the general toxicity of these estrogenic chemicals individually onClarias gariepinus. Fish were exposed to different concentrations of both NP and OP (250, 500, 750 and 1000 μg l−1) under semi‐static conditions for a period of 7 days. The adverse effect was evaluated with use of blood cell counting, hemoglobin (Hb), hematocrit (HCT), hematimetric indices, bilirubin, protein, glucose, serum transaminases, serum phosphatases, lactate dehydrogenase and cortisol. The results showed a clear indication of anemia, increases in leukocyte count and bilirubin content and a reduction in plasma protein levels with higher concentrations of both the toxicants compared with controls. Furthermore, with all the concentrations the inevitable increase in serum cortisol and plasma glucose showed primary and secondary stress responses. Moreover, probable tissue damage gave rise to a series of fluctuations of enzyme levels at lower concentrations, but a decrease with higher concentrations showed the severity of the effect. Depending on the parameters examined, OP had a relatively greater effect than NP. Overall, these two chemicals seemingly affected hematology and the activity of some enzymes, leading to serious impairment of the metabolism and physiology ofC.gariepinus. Copyright © 2011 John Wiley & Sons, Ltd.
Computational chemistry, systems biology and toxicology. Harnessing the chemistry of life: revolutionizing toxicology. A commentary
Authors: Kimber Ian , Humphris Colin , Westmoreland Carl , Alepee Nathalie , Negro Gianni Dal , Manou Irene
Published: March 2011

There is a continuing interest in, and increasing imperatives for, the development of alternative methods for toxicological evaluations that do not require the use of animals. Although a significant investment has resulted in some achievements, progress has been patchy and there remain many challenges. Among the most significant hurdles is developing non‐animal methods that would permit assessment of the potential for a chemical or drug to cause adverse health effects following repeated systemic exposure. Developing approaches to address this challenge has been one of the objectives of the European Partnership for Alternative Approaches to Animal Testing (EPAA). The EPAA is a unique partnership between the European Commission and industry that has interests in all aspects of reducing, refining and replacing the use of animals (the ‘3Rs’). One possible strategy that emerged from a broad scientific debate sponsored by the EPAA was the opportunity for developing entirely new paradigms for toxicity testing based upon harnessing the increasing power of computational chemistry in combination with advanced systems biology. This brief commentary summarizes a workshop organized by the EPAA in 2010, that had the ambitious title of ‘Harnessing the Chemistry of Life: Revolutionizing Toxicology’. At that workshop international experts in chemistry, systems biology and toxicology sought to map out how best developments in these sciences could be exploited to design new strategies for toxicity testing using adverse effects in the liver as an initial focus of attention. Here we describe the workshop design and outputs, the primary purpose being to stimulate debate about the need to align different areas of science with toxicology if new and truly innovative approaches to toxicity testing are to be developed. Copyright © 2011 John Wiley & Sons, Ltd.
Employment of single‐strand conformation polymorphism analysis in screening for α‐1,3 glucosyltransferase gene mutation A333V in Croatian population
Authors: Goreta Sandra Supraha , Dabelic Sanja , Dumic Jerka
Published: March 2011

Congenital disorder of glycosylation type Ic (CDG‐Ic) is caused by mutations in hALG6 gene encoding α‐1,3 glucosyltransferase (NP_037471.2), an enzyme that catalyzes the addition of the first glucose residue to the growing lipid‐linked oligosaccharide precursor in N‐glycosylation process. The most frequent mutation in hALG6 gene causing CDG‐Ic is c.998C>T that results in p.A333V substitution. Up‐to‐date, no CDG‐Ic patient has been detected in Croatia. However, as a part of the comprehensive project undertaken with the aim to estimate the frequencies of the carriers for specific mutations and polymorphisms related to particular CDGs in Croatian population, we screened genomic DNA samples obtained from 600 healthy nonconsanguineous Croatian residents to determine the frequency of the A333V mutation. For that purpose, we established the conditions for polymerase chain reaction‐based single‐strand conformation polymorphism analysis that is suitable for primary screening and in population studies, especially when the initial sample volume is small or DNA quantity is limited. None of the analyzed samples carried this mutation, indicating that the frequency of the patients carrying this homozygous mutation in Croatian population would be <1 in 1.4×106. J. Clin. Lab. Anal. 25:65;–70, 2011. © 2011 Wiley‐Liss, Inc.
Clinical applicability of single‐tube multiplex reverse‐transcriptase PCR in dengue virus diagnosis and serotyping
Authors: Mishra Bajayantimala , Sharma Mirnalini , Pujhari Sujit Kumar , Appannanavar Suma B. , Ratho Radha Kanta
Published: March 2011

This study has evaluated the clinical applicability of a single‐tube multiplex RT‐ PCR as compared with a two‐step nested RT‐PCR for the diagnosis as well as serotyping of dengue virus in patient's samples. Seventy‐six acute phase blood samples collected from clinically suspected dengue patients during the 2008 outbreak were subjected to two‐step nested RT‐PCR and single‐tube multiplex RT‐PCR for dengue diagnosis and serotyping. Of the 76 samples, 17 (22.4%) were positive for dengue viral RNA. Single dengue virus infection was found in 16 cases and 1 had concurrent infection with two serotypes (3&1). Dengue serotype 3 was the predominant serotype (70.5%), followed by serotype 1 (23.5%). Single‐tube multiplex PCR had concordant result with that of two‐step nested RT‐PCR including the one with concomitant infection. This study reveals the predominance of dengue serotype 3 in North India in addition to the co‐circulation of multiple serotypes and concomitant infection. The rapid and accurate diagnostic capability of single‐tube multiplex RT‐PCR used in the study appears to be promising enough to be commonly used for dengue viral detection as well as serotyping. J. Clin. Lab. Anal. 25:76–78, 2011. © 2011 Wiley‐Liss, Inc.
Comparison of RNA extraction methods to augment the sensitivity for the differentiation of vesicular stomatitis virus Indiana1 and New Jersey
Authors: Arshed Muhammad Javed , Magnuson Roberta J. , Triantis Joni , Abubakar Muhammad , Campen Hana Van , Salman Mo
Published: March 2011

Two methods for the extraction of RNA of vesicular stomatitis virus (VSV) Indiana1 and New Jersey and their simultaneous amplification by one‐step polymerase chain reaction using reverse transcriptase were evaluated. A guanidine‐thiocyanate‐based RNA extraction (Qiagen RNeasy Mini Kit, Qiagen, Valencia, CA ) followed by column‐based purification coupled with one‐step RT‐PCR proved to be a simple, safe, practicable, and reliable tool for rapid, highly sensitive, and specific differential diagnosis of both types of VSV in cell lysate and spiked tissue samples as compared with the tri‐phasic extraction method (Tri‐reagent method). When RNA was extracted either from VSV cell culture stock or from VSV spiked bovine lymph nodes by using Qiagen RNeasy Mini Kit, the detection limit in the multiplex RT‐PCR was as low as 0.505 to 2.84 TCID50 for VSV‐IND and VSV‐NJ, respectively. The multiplex RT‐PCR consistently detected VSV‐IND and NJ RNA in as little as 0.1–1.0 fg of total RNA from spiked BHK‐21 cell suspension when Qiagen RNeasy mini kit was used. The multiplex RT‐PCR assay was capable of detecting both types of VSV in a one‐step reaction tube. The minimum sensitivity of this assay in various experiments was 0.1683 TCID50 (IND), 0.0946 TCID50 (NJ), and 0.057 fg (IND and NJ) per 2 µl PCR sample, which is significantly more sensitive than reported previously (0.28–2.8 TCID50/1 µl). So the present study improved the sensitivity of previously reported multiplex RT‐PCR for the detection and differentiation of VSV‐IND and VSV‐NJ in a single assay. J. Clin. Lab. Anal. 25:95–99, 2011. © 2011 Wiley‐Liss, Inc.
Fetal RHD genotyping by analysis of maternal plasma in a mixed population
Authors: Amaral Daphne R.T. , Credidio Débora C. , Pellegrino Jordão , Castilho Lilian
Published: March 2011

Background: Maternal plasma analysis for the determination of the fetal RHD status is an exciting tool for the management of RhD‐negative pregnant women, specially sensitized women. We assessed the accuracy of fetal RHD genotyping by analysis of maternal plasma in a multi‐ethnic population. Methods: We analyzed plasma samples from 88 RhD‐negative pregnant women between 11 and 39 weeks of gestation, median age of 28 years old to determine the fetal RHD genotype. This population was from Southeastern Brazil with high mixed ethnic background. Fourteen patients (16%) had anti‐D alloantibody. We used Taqman primers and probes to detect by real‐time PCR, exons 4, 5, and 10 of RHD. As internal controls we used primers/probes sets to SRY and CCR5. Peripheral or umbilical cord bloods from respective nenonates were collected during delivery and hemagglutination was performed. Results: Fifty‐eight samples (66%) were genotyped as RHD+, 27 samples (31%) showed complete absence of RHD and 3 samples (3 %) presented a D variant (RHDψ). All the results agreed with the neonatal typing, including the three fetuses with the RHDψ, phenotyped as RhD‐negative. Thus, the accuracy of the fetal RHD genotyping in this mixed population was 100%. The earliest pregnancy in which fetal RHD was detected was 11 weeks. Conclusion: Our findings indicate that the accuracy of RHD gene using three regions (exons 4, 5, and 10) can be sufficient for clinical application in a multi‐ethnic population. This knowledge helped us on the development of a feasible protocol for fetal RHD genotyping on DNA from maternal plasma for our population. J. Clin. Lab. Anal. 25:100–104, 2011. © 2011 Wiley‐Liss, Inc.
Analysis of the temperature affects on leukocyte surface antigen expression
Authors: Jämsä Joel , Huotari Virva , Savolainen Eeva‐Riitta , Syrjälä Hannu , Ala‐Kokko Tero
Published: March 2011

Flow cytometric analysis of leukocyte surface antigens has been used to characterize infectious and septic processes in patients. We wanted to investigate how the sampling and processing temperature, the anticoagulant used, and the storage of the sample influence leukocyte immunophenotyping. Four blood samples, two using acid citrate dextrose and two using heparin as an anticoagulant, were taken from five intensive‐care unit patients with severe sepsis and five healthy volunteers. The samples were collected, stored, and processed either at +4°C or at room temperature (RT). The samples were processed for flow cytometric analysis within 1 hr of collection or after 6 or 24 hr storage. The surface antigens of interest were neutrophilic CD11b and CD64, monocytic CD11b, CD14, CD40, CD64, CD80 and HLA‐DR, and lymphocytic CD69 (separately in CD4+ and CD8+ T cells, B cells, and natural killer cells). The fluorescence intensities were higher at RT than at +4°C. During storage the intensities increased at RT, but at +4°C there were only minor changes. The effects were similar with both anticoagulants studied. According to our results, flow cytometric analysis of leukocyte surface antigen expressions should be performed using +4°C temperature throughout the process and within 6 hr. J. Clin. Lab. Anal. 25:118–125, 2011. © 2011 Wiley‐Liss, Inc.
Analytical quality goals for 25‐vitamin D based on biological variation
Authors: Viljoen Adie , Singh Dhruv K. , Farrington Ken , Twomey Patrick J.
Published: March 2011

Background: Measurement of 25‐hydroxyvitamin D, (25D) is central in the investigation of pathologies of bone and mineral ion metabolism and in determining a patient's vitamin D status. More recently much research interest has lead to investigating the role it can play in decreasing the risk of many chronic illnesses, including common cancers, autoimmune diseases, infectious diseases, and cardiovascular disease. Knowledge of the biological variation of an analyte forms an essential part of evaluating a new analyte enabling the objective assessment of the changes in serial results, the utility of reference intervals as well as establishing laboratory quality specifications. Methods: This study determined the biological variation of 25D in 20 healthy individuals that was calculated according to the familiar methods outlined by Fraser and Harris. Results: The within‐subject variation was 12.1% and the between subject variation was 40.3%. The critical difference for sequential values significant at P<0.05 was calculated as 38.4%. The within‐subject variation forms a relatively small part of the reference interval shown by the low index of individuality of 0.3. Objective analytical quality goals have also been established which have shown achievable minimum performance for imprecision of ∼6%. The desirable analytical bias goal was ∼10%. Conclusion: This study has objectively shown that the analytical precision of current instruments is being achieved contrary to the known problems surrounding the analytical bias for 25D assays. The limitations of using reference intervals for 25D, both in diagnoses and monitoring are shown. J. Clin. Lab. Anal. 25:130–133, 2011. © 2011 Wiley‐Liss, Inc.
Evaluation of saliva specimens as an alternative sampling method to detect hepatitis B surface antigen
Authors: Cruz Helena Medina , da Silva Elisangela Ferreira , Villela‐Nogueira Cristiane A. , Nabuco Letícia C. , Rodrigues do Ó Kycia Maria , Lewis‐Ximenez Lia Laura , Tachibana Yoshida Clara Fumiko , Lampe Elisabeth , Villar Livia Melo
Published: March 2011

In this study, a modified enzyme immunoassay (EIA) was evaluated for the Hepatitis B surface antigen (HBsAg) among whole saliva and oral fluid samples. Specimens were collected from 115 individuals who gave serum and oral fluid using Salivette (Sarstedt, Nümbrecht, Germany) and whole saliva. Saliva specimens were tested following a modified ELISA, and the results were compared with paired serum specimens that were tested according to the supplier's instructions. Transport buffer for the oral fluids, sample volume for assay, incubation period of sample with conjugate as well as cut‐off values were evaluated to optimize the assay. The highest sensitivity and specificity were obtained by increasing the incubation of sample and conjugate to 16 hr and using the area under the receiver operating characteristic curve to calculate cut‐off values. HBsAg was detected in 40 oral fluids and 44 whole saliva samples out of 47 paired positive serum specimens and not detected in 64 oral fluids and 63 whole saliva samples out of 68 matched negative sera samples by the ELISA assay. There was excellent agreement between the results for the serum and saliva specimens kappa value (κ): 0.80 for oral fluid and κ: 0.87 for whole saliva and there was excellent reproducibility. Using an optimized protocol, the sensitivities of whole saliva and oral fluid were 93.6 and 85.1%, respectively, whereas specificities of whole saliva and oral fluid were 92.6 and 94.1%, respectively. Our data showed a significant promise for the use of whole saliva and oral fluid together with the modified commercial EIA for Hepatitis B virus infection surveillance. J. Clin. Lab. Anal. 25:134–141, 2011. © 2011 Wiley‐Liss, Inc.
An efficient synthesis of pyrrolo[1,2‐a]quinazolin‐5(1H)‐one derivatives with the aid of low‐valent titanium reagent
Authors: Zhao Xuan , Shi Da‐Qing
Published: March 2011

A short and facile synthesis of 2,3,3a,4‐tetrahydropyrrolo[1,2‐a]quinazolin‐5(1H)‐ones and 2,3,3a,4‐tetrahydropyrrolo[1,2‐a]quinazoline‐1,5‐diones in good yields via the novel reductive cyclization of 2‐nitrobenzamides with haloketones or 4‐oxopentanoic acid promoted by low‐valent titanium reagent is described. J. Heterocyclic Chem., (2011).
Influence of internal fixator flexibility on murine fracture healing as characterized by mechanical testing and microCT imaging
Authors: Steck Roland , Ueno Masaki , Gregory Laura , Rijken Noortje. , Wullschleger Martin E. , Itoman Moritoshi , Schuetz Michael A.
Published: March 2011

Mechanically well‐defined stabilization systems have only recently become available, providing standardized conditions for studying the role of the mechanical environment on mouse bone fracture healing. The aim of this study was to characterize the time course of strength recovery and callus development of mouse femoral osteotomies stabilized with either low or high flexibility (in bending and torsion) internal fixation plates. Animals were euthanized and femora excised at 14, 21, and 28 days post‐osteotomy for microCT analysis and torsional strength testing. While a larger mineralized callus was observed in osteotomies under more flexible conditions at all time points, the earlier bridging of the mineralized callus under less flexible conditions by 1 week resulted in an earlier recovery of torsional strength in mice stabilized with low flexibility fixation. Ultimate torque values for these bones were significantly higher at 14 and 21 days post‐osteotomy compared to bones with the more flexible stabilization. Our study confirms the high reproducibility of the results that are achieved with this new implant system, therefore making it ideal for studying the influence of the mechanical environment on murine fracture healing under highly standardized conditions. © 2011 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 29: 1245–1250, 2011
Integrin‐associated tyrosine kinase FAK affects Cbfa1 expression
Authors: Schneider Galen B. , Zaharias Rebecca , Seabold Denise , Stanford Clark
Published: March 2011

Following cell adhesion, focal adhesion kinase (FAK) autophosphorylates on tyrosine and regulates intracellular signaling cascades that regulate cell growth and differentiation. The hypothesis of this study was FAK mediates osteoblast differentiation dependent Cbfa1 expression. Slowly mineralizing UI and rapidly mineralizing UMR‐106‐01 BSP osteoblasts formed focal adhesions; however, the level of FAK in UI focal adhesions was less than that seen in BSP cells. UI cultures had less FAK expression (p < 0.05) along with elevated levels of FAK phosphotyrosine in comparison to rapidly mineralizing BSP cultures. Mineralization decreased in a dose‐dependent manner in response to Herbimycin A, a tyrosine kinase inhibitor. Overexpression of FAK in UI cells led to a fourfold increase in Cbfa1 gene expression (p < 0.02), and an increase in Cbfa1 protein expression. These results suggest that the integrin‐associated tyrosine kinase FAK contributes to the regulation of the osteoblast differentiation in part through the regulation of Cbfa1 expression. © 2011 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 29: 1443–1447, 2011
Enhanced inactivation of bacteria by metal‐oxide nanoparticles combined with visible light irradiation
Authors: Lipovsky Anat , Gedanken Aharon , Nitzan Yeshayahu , Lubart Rachel
Published: March 2011

Background

In recent years nano‐metaloxides which easily penetrate into the cells with special interest due to their higher chemical reactivity as compared to that of similar materials in the bulk form. Of particular interest are nano‐TiO2 and ZnO, which have been widely used for their bactericidal and anticancerous properties.

Purpose

The aim of the present study was to examine the bactericidal properties of nano‐TiO2 and ZnO combined with visible light on S. aureus and S. epidermitis, known for their high prevalence in infected wounds.

Study

Using the technique of electron‐spin resonance (ESR) coupled with spin trapping, we examined the ability of TiO2 and ZnO nanoparticle suspensions in water to produce reactive oxygen species (ROS) with and without visible light irradiation. The possibility of exciting these nanoparticles with visible light in order to enhance their antimicrobial activity was also tested.

Results

Electron‐spin resonance measurements revealed that ZnO and TiO2 nanoparticles are able to produce ROS in water suspension. A remarkable enhancement of ROS production was found following illumination with blue light. In addition, illumination significantly enhanced the antibacterial activity of the nanoparticles.

Conclusion

The results suggest that nanoparticles combined with visible light can be used for sterilization purposes and may be effective for treating infected wounds. Lasers Surg. Med. 43:236–240, 2011. © 2011 Wiley‐Liss, Inc.
Global existence and blow‐up of solutions for nonlinear viscoelastic wave equation with degenerate damping and source
Authors: Han Xiaosen , Wang Mingxin
Published: March 2011

In this paper we investigate the global existence and finite time blow‐up of solutions to the nonlinear viscoelastic equation

associated with initial and Dirichlet boundary conditions. Here ∂j denote the sub‐differential of j. Under suitable assumptions on g(·), j(·) and the parameters in the equation, we obtain the global existence of generalized solutions, weak solutions for the equation. The finite time blow‐up of weak solutions for the equation is also established provided the initial energy is negative and the exponent p is greater than the critical value k + m. © 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim
Distance geometry in quasihypermetric spaces. III
Authors: Nickolas Peter , Wolf Reinhard
Published: March 2011

Let (X, d) be a compact metric space and let \documentclass{article}\usepackage{amssymb}\pagestyle{empty}\begin{document}$\mathcal {M}(X)$\end{document} denote the space of all finite signed Borel measures on X. Define \documentclass{article}\usepackage{amssymb}\pagestyle{empty}\begin{document}$I :\mathcal {M}(X)\longrightarrow \mathbb {R}$\end{document} by

and set $M(X) = \sup I(\mu ),$ where μ ranges over the collection of signed measures in \documentclass{article}\usepackage{amssymb}\pagestyle{empty}\begin{document}$\mathcal {M}(X)$\end{document} of total mass 1. This paper, with two earlier papers [Peter Nickolas and Reinhard Wolf, Distance geometry in quasihypermetric spaces. I and II], investigates the geometric constant M(X) and its relationship to the metric properties of X and the functional‐analytic properties of a certain subspace of \documentclass{article}\usepackage{amssymb}\pagestyle{empty}\begin{document}$\mathcal {M}(X)$\end{document} when equipped with a natural semi‐inner product. Specifically, this paper explores links between the properties of M(X) and metric embeddings of X, and the properties of M(X) when X is a finite metric space. © 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim © 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim
Understanding workplace deviant behavior in nonprofit organizations
Authors: Nair Nisha , Bhatnagar Deepti
Published: March 2011

Workplace deviant behavior has traditionally been studied with respect to business or for‐profit organizations. In this article, we argue that nonprofit organizations also experience deviance, and due to their unique characteristics, they deserve special attention for extending the understanding of workplace deviant behavior to other types of organizations. Based on a review of the literature on deviance, we develop a general model of workplace deviance that we apply to nonprofit organizations. Based on the integrative conceptual framework, we advance relevant propositions for understanding and explaining deviance in nonprofit organizations.
Knowledge, attitudes, and behaviors of nursing professionals and students in Beijing toward cardiovascular disease risk reduction
Authors: Wu Ying , Deng Ying , Zhang Ying
Published: March 2011

The rising incidence of cardiovascular disease (CVD) in China intensifies the need for effective health education for CVD risk reduction. The purpose of this study was to develop a description of nurses' knowledge about, attitudes toward, practice behaviors, and personal lifestyle behaviors related to CVD risk reduction. We surveyed 273 staff nurses, 35 nursing faculty, and 139 nursing students in Beijing. Most nurses could identify common risk factors for CVD and had positive attitudes toward CVD risk reduction. However, less than 58% of the respondents could correctly answer questions about evidence‐based recommendations for CVD risk reduction. This sample of Chinese nursing professionals and students lacked knowledge critical to providing guidance to individuals with or at risk for CVD. More intensive and creative approaches to the education of nursing professionals regarding CVD risk reduction are recommended. © 2011 Wiley Periodicals, Inc. Res Nurs Health 34:228–240, 2011
Formation of higher cyclic oligomers in polyamide‐4,6
Authors: Rulkens R. , Peters R.
Published: March 2011

An example of morphology‐steered chemistry of cyclic oligomers with the example of polyamide‐4,6 (PA46) is discussed in this article. The cyclic oligoamide profiles of solid‐state post condensed PA46 show an unusual maximum for intermediate cycles consisting of 10 and 12 amide groups, which are formed in the initial stage of solid state post condensation. This is explained from the short chain length at the start of the post condensation, the preorganization of linear oligomers in the lamellar crystal structure, and the occurrence of amide units in the PA46 lamellar folds. Further the development of the cyclic oligomers up to the cyclic hexadecamide is followed on heating the base polymer in the melt at 310 °C, where it showed to react back to the theoretical Jacobsen and Stockmayer distribution. © 2011 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem, 2011
Deposition of Titania‐containing Diamond‐like Carbon Nanocomposite Films by Sputtering‐assisted Chemical Vapor Deposition
Authors: Chen Kuo‐Cheng , Hong Franklin Chau‐Nan , Jeng Yeau‐Ren
Published: March 2011

Titania‐containing diamond‐like carbon (DLC) nanocomposite films were prepared by sputtering‐assisted plasma chemical vapor deposition. With titanium‐oxygen species sputtered from titania (TiO2) target by argon using a radio‐frequency (RF) power, DLC films were simultaneously grown on the negatively‐biased substrate by plasma chemical vapor deposition of acetylene gas using a pulsed direct‐current (DC) power. By adjusting the Ar/acetylene gas concentration, both TiO2 and TiC nanoparticles could be incorporated in the DLC films. TiO2 nanoparticles were mainly formed in the DLC matrix with TiO bond being the dominant bonding for Ti in the DLC films. Furthermore, the amount of TiO2 and TiC nanoparticles embedded in the DLC film increased with the increase of Ar concentration in the gas. The TiO2‐DLC nanocomposite films deposited at 80% Ar exhibited a high hardness of around 13 GPa at a relatively low stress and, particularly, a fast rate of turning super‐hydrophilic by reaching zero degree of water contact angle under 40 min of ultraviolet irradiation.

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Carbon Nanoparticle/Hydrogenated Amorphous Carbon Composite Thin Films Formed in ECR Plasma
Authors: Calafat Maria , Yuryev Pavel , Drenik Aleksander , Slim Aref , Clergereaux Richard
Published: March 2011

Composite thin films are deposited from acetylene in a microwave multipolar plasma excited at distributed electron cyclotron resonance in one single process. The composite consists of carbon‐based nanoparticles embedded in a hydrogenated amorphous matrix. Effects of plasma duration and microwave power on the composite thin film are described along with the powder growth mechanisms. Indeed, two types of nanoparticles are formed: pure graphite‐like and graphite‐like shell with a metallic core. The first type grows from recombinations in the plasma phase, while core‐shelled starts from a metallic core generated from the reactor walls. These metallic clusters can be heated while immersed in the plasma which leads to catalytic growth of powders.

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