bluuebunny/crossref_metadata_2025_split huggingface dataset:
First construction of infectious clone for newly emerging mutation porcine circovirus type 2 (PCV2) followed by comparison with PCV2a and PCV2b genotypes in biological characteristics in vitro
Authors: Guo Long J , Lu Yue H , Huang Li P , Wei Yan W , Wu Hong L , Liu Chang M 
Published: June 2011

Abstract

Background
Porcine circovirus type 2 (PCV2), the causative agent of postweaning multisystemic wasting syndrome (PMWS), is a serious economic problem in the swine industry. Different genotypes (PCV2a, PCV2b and PCV2d) of the virus are present in the clinical cases in China, and it is necessary to elucidate the pathogenic difference among different genotypes of PCV2. In this study, four strains of different genotypes were isolated, two were ordinary strains and another two were mutation strains, which there are one and two amino acids elongation in the capsid protein (Cap) of PCV2, respectively. Representative strains of different genotypes of the virus were constructed by infectious molecular clone and biological characterization of the rescued viruses were identified in vitro.

Results
Four PCV2 isolates (PCV2a/CL, PCV2b/YJ, PCV2b/JF and PCV2d/BDH) of different genotypes were isolated from the clinical cases of PMWS in China. Four infectious clones of PCV2 were constructed and the rescued viruses were harvested after transfection into PK15 cells. The rescued viruses were verified by nucleotide sequence analysis, morphology of the viruses and immunoperoxidase monolayer assay (IPMA). The rescued viruses propagated stably after consecutive incubation for more than ten passages, and virus propagation reached its peak 72h post infection (PI), and the virus titers were up to 105.7 TCID50/ml. By using neutralizing 1D2 monoclonal antibody (mAb) of PCV2, the antigen capture ELISA showed that only the PCV2a/rCL and PCV2b/rJF strains has immunoreactivity with the 1D2 mAb, however, another two rescued strains (PCV2b/rYJ and PCV2d/rBDH) do not, which indicated the antigenic difference among the rescued viruses of different genotypes. In addition, here is the first report of obtaining the newly emerging PCV2 with mutation in vitro by infectious molecular clone technology.

Conclusions
Conclusions drawn from this study show that PCV2 has prevailing differences in genomic and ORF2 gene length and antigen in swine herds in China. Four representative clones for different genotypes were constructed and rescued, which will facilitate further studies on the pathogenic differences resulting from different subtypes of PCV2.
Effective inhibition of foot-and-mouth disease virus (FMDV) replication in vitro by vector-delivered microRNAs targeting the 3D gene
Authors: Du Junzheng , Gao Shandian , Luo Jihuai , Zhang Guofeng , Cong Guozheng , Shao Junjun , Lin Tong , Cai Xuepeng , Chang Huiyun 
Published: June 2011

Abstract
Background
Foot-and-mouth disease virus (FMDV) causes an economically important and highly contagious disease of cloven-hoofed animals. RNAi triggered by small RNA molecules, including siRNAs and miRNAs, offers a new approach for controlling viral infections. There is no report available for FMDV inhibition by vector-delivered miRNA, although miRNA is believed to have more potential than siRNA. In this study, the inhibitory effects of vector-delivered miRNAs targeting the 3D gene on FMDV replication were examined.

Results
Four pairs of oligonucleotides encoding 3D-specific miRNA of FMDV were designed and selected for construction of miRNA expression plasmids. In the reporter assays, two of four miRNA expression plasmids were able to significantly silence the expression of 3D-GFP fusion proteins from the reporter plasmid, p3D-GFP, which was cotransfected with each miRNA expression plasmid. After detecting the silencing effects of the reporter genes, the inhibitory effects of FMDV replication were determined in the miRNA expression plasmid-transfected and FMDV-infected cells. Virus titration and real-time RT-PCR assays showed that the p3D715-miR and p3D983-miR plasmids were able to potently inhibit the replication of FMDV when BHK-21 cells were infected with FMDV.

Conclusion
Our results indicated that vector-delivered miRNAs targeting the 3D gene efficiently inhibits FMDV replication in vitro. This finding provides evidence that miRNAs could be used as a potential tool against FMDV infection.
A conserved juxtacrine signal regulates synaptic partner recognition in Caenorhabditis elegans
Authors: Park Joori , Knezevich Philip Louis , Wung William , O'Hanlon Shanté Nicole , Goyal Akshi , Benedetti Kelli Leilani , Barsi-Rhyne Benjamin James , Raman Mekala , Mock Natalyn , Bremer Martina , VanHoven Miri Kerensa 
Published: June 2011

Abstract

Background
An essential stage of neural development involves the assembly of neural circuits via formation of inter-neuronal connections. Early steps in neural circuit formation, including cell migration, axon guidance, and the localization of synaptic components, are well described. However, upon reaching their target region, most neurites still contact many potential partners. In order to assemble functional circuits, it is critical that within this group of cells, neurons identify and form connections only with their appropriate partners, a process we call synaptic partner recognition (SPR). To understand how SPR is mediated, we previously developed a genetically encoded fluorescent trans-synaptic marker called NLG-1 GRASP, which labels synaptic contacts between individual neurons of interest in dense cellular environments in the genetic model organism Caenorhabditis elegans.

Results
Here, we describe the first use of NLG-1 GRASP technology, to identify SPR genes that function in this critical process. The NLG-1 GRASP system allows us to assess synaptogenesis between PHB sensory neurons and AVA interneurons instantly in live animals, making genetic analysis feasible. Additionally, we employ a behavioral assay to specifically test PHB sensory circuit function. Utilizing this approach, we reveal a new role for the secreted UNC-6/Netrin ligand and its transmembrane receptor UNC-40/Deleted in colorectal cancer (DCC) in SPR. Synapses between PHB and AVA are severely reduced in unc-6 and unc-40 animals despite normal axon guidance and subcellular localization of synaptic components. Additionally, behavioral defects indicate a complete disruption of PHB circuit function in unc-40 mutants. Our data indicate that UNC-40 and UNC-6 function in PHB and AVA, respectively, to specify SPR. Strikingly, overexpression of UNC-6 in postsynaptic neurons is sufficient to promote increased PHB-AVA synaptogenesis and to potentiate the behavioral response beyond wild-type levels. Furthermore, an artificially membrane-tethered UNC-6 expressed in the postsynaptic neurons promotes SPR, consistent with a short-range signal between adjacent synaptic partners.

Conclusions
These results indicate that the conserved UNC-6/Netrin-UNC-40/DCC ligand-receptor pair has a previously unknown function, acting in a juxtacrine manner to specify recognition of individual postsynaptic neurons. Furthermore, they illustrate the potential of this new approach, combining NLG-1 GRASP and behavioral analysis, in gene discovery and characterization.
High prevalence of HIV‐1 drug resistance among patients on first‐line antiretroviral treatment in Lomé, Togo
Authors: Dagnra Anoumou Y , Vidal Nicole , Mensah Akovi , Patassi Akouda , Aho Komi , Salou Mounerou , Monleau Marjorie , Prince‐David Mireille , Singo Assétina , Pitche Palokinam , Delaporte Eric , Peeters Martine 
Published: June 2011

BackgroundWith widespread use of antiretroviral (ARV) drugs in Africa, one of the major potential challenges is the risk of emergence of ARV drug‐resistant HIV strains. Our objective is to evaluate the virological failure and genotypic drug‐resistance mutations in patients receiving first‐line highly active antiretroviral therapy (HAART) in routine clinics that use the World Health Organization public health approach to monitor antiretroviral treatment (ART) in Togo.MethodsPatients on HAART for one year (10‐14 months) were enrolled between April and October 2008 at three sites in Lomé, the capital city of Togo. Plasma viral load was measured with the NucliSENS EasyQ HIV‐1 assay (Biomérieux, Lyon, France) and/or a Generic viral load assay (Biocentric, Bandol, France). Genotypic drug‐resistance testing was performed with an inhouse assay on plasma samples from patients with viral loads of more than 1000 copies/ml. CD4 cell counts and demographic data were also obtained from medical records.ResultsA total of 188 patients receiving first‐line antiretroviral treatment were enrolled, and 58 (30.8%) of them experienced virologic failure. Drug‐resistance mutations were present in 46 patients, corresponding to 24.5% of all patients enrolled in the study. All 46 patients were resistant to non‐nucleoside reverse‐transcriptase inhibitors (NNRTIs): of these, 12 were resistant only to NNRTIs, 25 to NNRTIs and lamivudine/emtricitabine, and eight to all three drugs of their ARV regimes. Importantly, eight patients were already predicted to be resistant to etravirine, the new NNRTI, and three patients harboured the K65R mutation, inducing major resistance to tenofovir.ConclusionsIn Togo, efforts to provide access to ARV therapy for infected persons have increased since 2003, and scaling up of ART started in 2007. The high number of resistant strains observed in Togo shows clearly that the emergence of HIV drug resistance is of increasing concern in countries where ART is now widely used, and can compromise the long‐term success of first‐ and second‐line ART.
Epithelial to mesenchymal transition markers expressed in circulating tumour cells of early and metastatic breast cancer patients
Authors: Kallergi Galatea , Papadaki Maria A , Politaki Eleni , Mavroudis Dimitris , Georgoulias Vassilis , Agelaki Sophia 
Published: June 2011

Abstract
Introduction
Epithelial to mesenchymal transition (EMT) is considered an essential process in the metastatic cascade. EMT is characterised by upregulation of vimentin, Twist, Snail, Slug and Sip1 among others. Metastasis is also associated with the presence of circulating tumour cells (CTCs) and disseminated tumour cells in the blood and bone marrow, respectively, of breast cancer patients, but the expression of EMT markers in these cells has not been reported so far.

Methods
The expression of Twist and vimentin in CTCs of 25 metastatic and 25 early breast cancer patients was investigated by using double-immunofluorescence experiments in isolated peripheral blood mononuclear cell cytospins using anti-cytokeratin (anti-CK) anti-mouse (A45-B/B3) and anti-Twist or anti-vimentin anti-rabbit antibodies.

Results
Among early breast cancer patients, vimentin-and Twist-expressing CK+ CTCs were identified in 77% and 73% of the patients, respectively, and in 100% of the patients with metastatic breast cancer for both markers (P = 0.004 and P = 0.037, respectively). Among patients with early disease, 56% and 53% of the CK+ CTCs were double-stained with vimentin and Twist, and the corresponding values for metastatic patients were 74% and 97%, respectively (P = 0.005 and P = 0.0001, respectively). The median expression of CK+vimentin+ and CK+Twist+ cells per patient in metastatic patients was 98% and 100%, and in an adjuvant chemotherapy setting the corresponding numbers were 56% and 40.6%, respectively. Triple-staining experiments revealed that all CK+Twist+ or CK+vimentin+ cells were also CD45-, confirming their epithelial origin. Immunomagnetic separation of CTCs and triple-immunofluorescence with anti-CK/anti-Twist/anti-vimentin antibodies demonstrated that both mesenchymal markers could be coexpressed in the same CK+ cell, since 64% of the total identified CTCs were triple-stained. There was a significant correlation (P = 0.005) between the number of CTCs expressing Twist and vimentin within the same setting.

Conclusions
CTCs expressing Twist and vimentin, suggestive of EMT, are identified in patients with breast cancer. The high incidence of these cells in patients with metastatic disease compared to early stage breast cancer strongly supports the notion that EMT is involved in the metastatic potential of CTCs.
Smoking Cessation Practices of Dental Care Providers in Eastern Province of Saudi Arabia
Authors: Subaie Ahmad E. , Nasser Muhsen M. , Qussair Ali A. , Ali Aiman A. , Jandan Badr A. Al 
Published: June 2011

AbstractBackground:Awareness programs are highly recommended to educate smokers and help them to quit. The aim of this study is to evaluate dental professionals' awareness and utilisation of the 5As in the Eastern Province of Saudi Arabia. Material andMethods:A survey was conducted in Saudi Arabia on 102 dental care providers (DCPs) in 2010 to assess awareness of 5As. Many factors were included in the survey such as demographic characteristics, working place, years of experiences, level of education and country of graduation.Results:Only 23 (22.5%) of our sample know about the 5As (Ask, Advise, Assess, Assist and Arrange), and less than 7% of DCPs assist their patients to quit smoking. In addition, 19 (18.6%) of DCPs were smokers, and of them 6 (31.6%) smoked in front of their patients.Conclusion:Most of the dental practitioners were not aware of the 5As for smoking cessation. Most of them they did not pay attention to the willingness of their patients to quit smoking.
Client Satisfaction With English Stop Smoking Services
Authors: May Sylvia , McEwen Andy 
Published: June 2011

AbstractThis study reports client satisfaction in five UK National Health Service (NHS) Stop Smoking Services (SSS) and examines the impact of satisfaction on outcome four weeks postquit. Six services were selected on the basis of perceived ability to run the study from a pool of 11 who responded to a mailshot to volunteer. Advisors made one of two satisfaction surveys available to their clients two weeks after quitting smoking. The two surveys were a ‘brief’ three-item survey and a ‘full’ survey containing those items plus 19 others about specific aspects of the service. There were two key items: how satisfied respondents were with the support they received to stop smoking and would they recommend the service to another smoker. Surveys were completed anonymously but had unique identification numbers so they could be linked to individual demographic and outcome data. Overall 10% (554/5520) of treated smokers in the services completed a survey, 7% (390/5520) completed the ‘full’ survey. Responses were a strong endorsement of the participating services, with 93% (505/542) satisfied or very satisfied with the service and 99% (548/552) prepared to recommend the service. The sample was unrepresentative of the population of treated smokers in terms of eligibility for free prescriptions, ethnicity, age, type of treatment experienced, the type of advisor seen, medication use and end of treatment outcome. Satisfaction at week two was not related to abstinence at four weeks postquit. Limitations of the study and suggestions for service delivery are discussed.
Dual polar spaces of arbitrary rank
Authors: Huggenberger Simon 
Published: June 2011

Abstract
In 1982 P. Cameron gave a characterisation of dual polar spaces of finite rank viewed as point-line spaces. This characterisation makes essential use of the fact that dual polar spaces of finite rank have finite diameter. Our goal is to give a characterisation which includes dual polar spaces of infinite rank. Since dual polar spaces of infinite rank are disconnected, we introduce a point-relation that denotes pairs of points at “maximal distance”, and we call this an opposition relation. This approach is in the spirit of the theory of twin buildings.
Isolation of dipeptidyl peptidase IV (DP 4) isoforms from porcine kidney by preparative isoelectric focusing to improve crystallization
Authors: Wagner Leona , Wermann Michael , Rosche Fred , Rahfeld Jens-Ulrich , Hoffmann Torsten , Demuth Hans-Ulrich 
Published: June 2011

AbstractIn the present studies we resolved the post-translational microheterogeneity of purified porcine dipeptidyl peptidase IV (DP 4) from kidney cortex. Applying SDS-homogeneous DP 4 onto an analytical agarose isoelectric focusing (IEF) gel, pH 4–6, activity staining resulted in at least 17 isoforms between pH 4.8–6.0. These could be separated into fractions with only two to six isoforms by means of preparative liquid-phase IEF, using a Rotofor cell. Starting off with three parallel Rotofor runs under the same conditions at pH 5–6, the fractions were pooled according to the specific activity of DP 4, pH and analytical IEF profile, and further refractionated without any additional ampholytes. Since excessive dilution of ampholytes and proteins was kept to the minimum, a second refractionation step could be introduced, resulting in pH gradients between 0.022 and 0.028 pH increments per fraction. By performing two consecutive refractionation steps, the high resolution necessary for the separation of DP 4 isoforms could be achieved. This represents an alternative method if isolation of isoforms with similar pI's results in precipitation and denaturation in presence of a narrow pH range. Furthermore, it demonstrates that preparative IEF is a powerful tool to resolve post-translational microheterogeneity of a purified protein required for crystallization processing.
Bio based fuels and fuel additives from lignocellulose feedstock via the production of levulinic acid and furfural
Authors: Dautzenberg Geertje , Gerhardt Mirko , Kamm Birgit 
Published: June 2011

Abstract
The demand for biomass-derived fuels and fuel additives, particularly in the transportation sector, has stimulated intense research efforts in the chemistry of levulinic acid and levulinic acid secondary products over the past decade. Additionally, recent technological progress in lignocellulosic feedstock (LCF) chemistry has also increased attention in this regard. As a result, several oxygenating fuel additives with potential applications in both gasoline and diesel fuels have been identified. Some of the chemicals, such as ethyl valerate, appear to be viable alternatives to the currently used branched, short-chain ethers that are derived from side products of petrol refining. Cost-effective applications of lignocellulosic biomass are a crucial aspect of its feasibility. In consideration of the LCF biorefinery concept, the feasibility must also include the chemical pulping of LCF and the comprehensive utilisation of its main constituents cellulose, hemicelluloses, and lignin. The present study focuses on cellulose and hemicelluloses as viable sources for the production of biofuels and biofuel additives. Multifunctional catalysis, including hydrogenation and acid catalysis are the primary instruments used for the conversion of the monomeric carbohydrate building blocks, i.e., mainly C5 sugars, such as xylose and arabinose, and C6 sugars in the form of glucose and their respective secondary products, furfural and levulinic acid. Lignin utilisation is not addressed in this paper.
Lignin oxidation mechanisms under oxygen delignification conditions. Part 1. Results from direct analyses
Authors: Kalliola Anna , Kuitunen Susanna , Liitiä Tiina , Rovio Stella , Ohra-aho Taina , Vuorinen Tapani , Tamminen Tarja 
Published: June 2011

AbstractOxidation of softwood and hardwood kraft lignins was observed under conditions of oxygen delignification (90°C and 110°C; 0.6 and 0.9 MPa) as a function of time by means of a number of analysis techniques and quantitative information was obtained on the degradation and formation of various compounds and structures. The decrease in reactor pressure was monitored during a 4-h reaction period. During the first 60 min, lignin reactivity was high, while a very intense stage took place during the first 20 min. The reactions decelerated after the first 60 min and after 120 min the reactions did not significantly advance. The oxygen consumption after 4-h reaction was 1.3–1.5 mole O2per 1 mole lignin depending on the conditions. In the first 20 min, 50%–60% of the oxygen was consumed and the consumption increased only slightly after 60 min. At 90°C, the changes in all observed quantities were smaller throughout the whole 4-h reaction period than at 110°C. Under the studied conditions, increasing the reaction temperature, rather than the pressure, had primary significance in the increasing rate of lignin degradation. Hardwood kraft lignin was more reactive than softwood kraft lignin. The results obtained in this study are the basis for the development of a mechanistic model for the oxygen delignification process of pulps to be published in subsequent papers.
Effects of loading adjustment on the reliability degradation of composites
Authors: Chen Chung-Ling , Wang Kuo-Shong 
Published: June 2011

AbstractThis paper defines two parameters to describe the effects of fatigue load adjustment on the reliability degradation of composites. The transition period, n2a, indicates the free failure time period from the instant of high-low adjustment until the residual strength distribution meets the low-level stress. The reliability drop, ∣ΔR∣, is shown at the instant of low-high adjustment. Based on the strength-life equal rank assumption, a typical Yang model of the residual strength is considered to derive these two items by retrieving the residual strength distributions to the corresponding initials. The predicted parameters are verified to be in good agreement with the Monte Carlo simulation of fatigue failure. It is found that n2a increases dramatically as the reliability Ra at which the stress level changes, in the range of 1>Ra>0.95. The relationship between ∣ΔR∣ and Ra exhibits a parabolic-like curve with a peak near the mean cycles to failure of the process at low-level stress.
Strengthening of reinforced concrete corbels with GFRP overlays
Authors: Ozden Sevket , Atalay Hilal Meydanli 
Published: June 2011

AbstractThe strength and post-peak performance of reinforced concrete corbels, strengthened with epoxy bonded glass fiber reinforced polymer (GFRP) overlays, were experimentally investigated. The test variables were the corbel shear span to depth ratio, corbel main reinforcement ratio, and the number and orientation of the GFRP fibers. In total, 24 normal strength concrete, one-third scale, corbel specimens, without hoop reinforcement, were tested to failure under quasi-static gravity loading. Test results revealed that GFRP overlays can easily be used for the enhancement of corbel load bearing capacity, depending on the fiber orientation. The main reinforcement ratio and the number of GFRP plies were found to be the two main variables affecting the level of strength gain in the corbel specimens.