bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparative studies of the capsid precursor polypeptide P1 and the capsid protein VP1 cDNA vectors for DNA vaccination against foot‐and‐mouth disease virus
Authors: Yang Ning‐Sun , Wang Jeng‐Hwan , Lin Ku‐Feng , Wang Chien‐Yu , Kim Suk‐Am , Yang Yu‐Ling , Jong Ming‐Hwa , Kuo Tsun‐Yung , Lai Shiow‐Suey , Holland Cheng R. , Chan Ming‐Tsair , Liang Shu‐Mei 
Published: February 2005

AbstractBackgroundFoot‐and‐mouth disease virus (FMDV) causes a severe livestock disease, and the virus is an interesting target for virology and vaccine studies.Materials and methodsHere we evaluated comparatively three different viral antigen‐encoding DNA sequences, delivered via two physical means (i.e., gene gun delivery into skin and electroporation delivery into muscle), for naked DNA‐mediated vaccination in a mouse system.ResultsBoth methods gave similar results, demonstrating commonality of the observed DNA vaccine effects. Immunization with a cDNA vector expressing the major viral antigen (VP1) alone routinely failed to induce the production of anti‐VP1 or neutralizing antibodies in test mice. As a second approach, the plasmid L‐VP1 that produces a transgenic membrane‐anchored VP1 protein elicited a strong antibody response, but all test mice failed in the FMDV challenge experiment. In contrast, for mice immunized with the viral capsid precursor protein (P1) cDNA expression vector, both neutralizing antibodies and 80–100% protection in test mice were detected.ConclusionsThis strategy of using the whole capsid precursor protein P1 cDNA for vaccination, intentionally without the use of virus‐specific protease or other encoding genes for safety reasons, may thus be employed as a relevant experimental system for induction or upgrading of effective neutralizing antibody response, and as a convenient surrogate test system for DNA vaccination studies of FMDV and presumably other viral diseases. Copyright © 2005 John Wiley & Sons, Ltd.
Generation of a packaging cell line for prolonged large‐scale production of high‐titer HIV‐1‐based lentiviral vector
Authors: Ni Yajin , Sun Susan , Oparaocha Ibe , Humeau Laurent , Davis Brian , Cohen Reuben , Binder Gwendolyn , Chang Yung‐Nien , Slepushkin Vladimir , Dropulic Boro 
Published: February 2005

AbstractBackgroundAn Erratum has been published for this article in Journal of Gene Medicine 7(6), 2005, 835.A stable packaging cell line facilitates large‐scale lentivirus vector manufacture. However, it has been difficult to produce clinical‐scale HIV‐1‐based lentiviral vectors using a packaging cell line, in part due to toxicity of packaging genes, and gene silencing that occurs during the long culture period necessary for sequential addition of packaging constructs.MethodsTo avoid these problems, we developed a three‐level cascade gene regulation system designed to remove tetracycline transactivator (tTA) from cytomegalovirus immediate early promoter (CMV)‐controlled expression to reduce cytotoxicity from constitutive expression of tTA and leaky expression of packaging genes. We also performed a one‐step integration of the three packaging plasmids to shorten the culture time for clonal selection.ResultsAlthough leaky expression of p24 and vector production still occurred despite the three‐level regulation system, little cytotoxicity was observed and producer cells could be expanded for large‐scale production. Producer cells yielded remarkably stable vector production over a period greater than 11 days with the highest titer 3.5 × 107 transducing units (TU)/ml and p24 300 ng/ml, yielding 2.2 × 1011 TU and 1.8 milligram (mg) p24 from one cell factory. No replication‐competent lentivirus (RCL) was detected. Long‐term analysis demonstrated that, although the cells are genetically stable, partial gene silencing occurs after 2–3 months in culture; however, the one‐step construct integration allowed prolonged vector production before significant gene silencing. Concentrated vector resulted in 90% transduction in CD4+ lymphocytes at 20 TU per cell. CD34+ progenitor cells were transduced at 41–46% efficiency, and long‐term initiating culture (LTC‐IC) was transduced at 45–51%.ConclusionsThese results demonstrate for the first time HIV‐1‐based lentiviral vector production on the large scale using a packaging cell line. Copyright © 2005 John Wiley & Sons, Ltd.
P21‐activated kinase‐1 is necessary for depolarization‐mediated neuronal survival
Authors: Johnson Kyle , D'Mello Santosh R. 
Published: February 2005

AbstractCerebellar granule neurons undergo apoptosis when switched from culture medium containing high potassium (HK) to medium that contains low potassium (LK). HK treatment leads to an activation of p21‐activated kinase‐1 (PAK‐1). Overexpression of a constitutively active form of PAK‐1 protects against apoptosis in LK medium. Overexpression of a dominant‐negative form of PAK‐1 blocks survival in HK. Although PAK‐1 is usually considered to be a downstream effector of Rac and Cdc42, we were unable to detect association between PAK‐1 and either Rac1 or Cdc42 in cerebellar granule neurons. Interaction between PAK‐1 and PDK1 is detected in granule neurons, although there is no change in the extent of interaction in neurons primed to die. Neuronal survival by PAK‐1 overexpression is not inhibited by PD98059 or LY294002, which inhibit the activity of MEK and PI‐3 kinase, respectively. The ability of PAK‐1 to maintain neuronal survival is, however, blocked by ML‐9, a compound known to inhibit Akt. Our results show that that PAK‐1 is necessary for neuronal survival in HK and suggest that its neuroprotective action may be mediated by a GTPase‐independent, but Akt‐dependent, mechanism. © 2005 Wiley‐Liss, Inc.
Psychological success: When the career is acalling
Authors: Hall Douglas T. , Chandler Dawn E. 
Published: February 2005

AbstractThis article has the dual purpose of expanding an understanding of the relationship between subjective and objective careers, and describing one condition under which the subjective career takes on particular salience: when the person feels a sense of calling in his or her career (that is, a sense of purpose, that this is the work one was meant to do.) This sense of calling does not necessarily have to be connected to a set of religious beliefs. We present a model of psychological success based on the career as a calling in order to clarify relationships between the subjective and objective career, and we offer propositions related to the model. Further, we offer a case study to illustrate the notion of the career as a calling, as proposed in the model. Copyright © 2005 John Wiley & Sons, Ltd.
Evaluating methods for the reliability of a large 2‐dimensional rectangular k‐within‐consecutive‐(r, s)‐out‐of‐(m, n):F system
Authors: Yamamoto Hisashi , Akiba Tomoaki 
Published: February 2005

AbstractA 2‐dimensional rectangular k‐within‐consecutive‐(r, s)‐out‐of‐(m, n):F system consists of m × n components, and fails if and only if k or more components fail in an r × s submatrix. This system can be treated as a reliability model for TFT liquid crystal displays, wireless communication networks, etc. Although an effective method has been developed for evaluating the exact system reliability of small or medium‐sized systems, that method needs extremely high computing time and memory capacity when applied to larger systems. Therefore, developing upper and lower bounds and accurate approximations for system reliability is useful for large systems. In this paper, first, we propose new upper and lower bounds for the reliability of a 2‐dimensional rectangular k‐within‐consecutive‐(r, s)‐out‐of‐(m, n):F system. Secondly, we propose two limit theorems for that system. With these theorems we can obtain accurate approximations for system reliabilities when the system is large and component reliabilities are close to one. © 2005 Wiley Periodicals, Inc. Naval Research Logistics, 2005
Shock model in Markovian environment
Authors: Li Gang , Luo Jiaowan 
Published: February 2005

AbstractA Markov modulated shock models is studied in this paper. In this model, both the interarrival time and the magnitude of the shock are determined by a Markov process. The system fails whenever a shock magnitude exceeds a pre‐specified level η. Nonexponential bounds of the reliability are given when the interarrival time has heavy‐tailed distribution. The exponential decay of the reliability function and the asymptotic failure rate are also considered for the light‐tailed case. © 2005 Wiley Periodicals, Inc. Naval Research Logistics, 2005
Regime change and nation building: can donors restore governance in post‐conflict states?
Authors: Rondinelli Dennis A. , Montgomery John D. 
Published: February 2005

AbstractForeign aid agencies and international assistance organisations are now heavily involved in nation building in post‐conflict states. Their record of strengthening democratic governance in countries where civil war or military force replaced unpopular regimes is mixed. Experience suggests that a complex set of conditions must be created quickly in order to rebuild indigenous governance. Ensuring security, providing assistance through a transparent and coherent plan of action, coordinating donors' activities, establishing strong and legitimate national authority, strengthening democratic political processes, transferring responsibility and resources for development to a new government, stabilising the economy and strengthening social capital and human assets must all be done in quick succession. Achieving these goals requires a cadre of civilian and, sometimes, military personnel with expertise in post‐conflict nation building. The frequency with which government aid programmes and international assistance organisations engage in post‐conflict reconstruction also suggests the need for more explicit national and international policies and the creation of specialised nation‐building agencies to undertake these difficult tasks. Copyright © 2005 John Wiley & Sons, Ltd.
Strengthening democratic governance of the security sector in conflict‐affected countries
Authors: Ball Nicole 
Published: February 2005

AbstractSecurity for people, communities and states is essential for sustainable development, democratisation and conflict mitigation. Politicised, badly managed or ineffective security bodies and justice systems often create instability and insecurity, largely due to the lack of effective democratic systems. Strengthening democratic security‐sector governance after conflict presents enormous challenges, particularly: (1) developing and implementing a legal framework consistent with international law and democratic practice; (2) developing effective, well‐functioning civil management and oversight bodies; (3) developing viable, accountable and affordable security forces; (4) ensuring that the institutional culture of the security forces supports the legal framework, international law, good democratic practice and civil management and oversight bodies. Addressing these challenges requires professional security forces, capable civil authorities, rule of law and regional approaches. Reform activities should be guided by local ownership, sensitivity to the politics of reform, local capacity, local context and a comprehensive sector‐wide framework. Local stakeholders must make hard decisions about priorities on the availability of domestic resources available and the costs and benefits of accepting external assistance. Copyright © 2005 John Wiley & Sons, Ltd.
Decentralisation, democracy and development in a post‐conflict society: commune councils in Cambodia
Authors: Blunt Peter , Turner Mark 
Published: February 2005

AbstractThis article explores explanations for the development of a particular form of decentralisation in the post‐conflict state of Cambodia. It looks at the context in which decentralisation has taken place, and analyses critical aspects of the functioning of the main elements of decentralisation: the commune councils. The article demonstrates that decentralisation has faltered due to a lack of fit with Cambodia's socio‐cultural and institutional context. This helps to explain why there has been relatively little devolution of decision‐making power to commune councils in Cambodia. But this is not the complete answer. A more powerful explanation is one in which an unconducive general environment for decentralisation complements a lack of real political enthusiasm for the idea, and a government agenda that is more consistent with pragmatic short‐term political gains (such as the consolidation of political party interests) than it is with the bolder, largely ideologically‐driven interests of donors in the post‐conflict establishment of strong forms of popular participation and political pluralism. Copyright © 2005 John Wiley & Sons, Ltd.
Perspectives in spicing up proteomics with splicing
Authors: Godovac‐Zimmermann Jasminka , Kleiner Oliver , Brown Larry R. , Drukier Andrzej K. 
Published: February 2005

AbstractIn the post‐genomics era there has been an acceleration of understanding of cellular and organismal biology and this acceleration has moved the goalposts for proteomics. Higher eukaryotes use alternative promoters, alternative splicing, RNA editing and post‐translational modification to produce multiple isoforms of proteins from single genes. Switching amongst these isoforms is a major mechanism for control of cellular function. At present fundamental limitations in sensitivity, in absolute quantitation of proteins and in the characterization of protein structure at functionally important levels strongly limit the applicability of proteomics to higher eukaryotes. Recent developments suggest that quantitative, top‐down proteomics analyses of complete proteins at sub‐attomole levels are necessary for physiologically relevant studies of higher eukaryotes. New proteomics technologies which will ensure the future of proteomics as an important technology in medicine and cellular biology of higher eukaryotes are becoming available.
A new application of microwave technology to proteomics
Authors: Juan Hsueh‐Fen , Chang Shing‐Chuan , Huang Hsuan‐Cheng , Chen Shui‐Tein 
Published: February 2005

AbstractTwo‐dimensional electrophoresis (2‐DE) combined with mass spectrometry has significantly improved the possibilities of large‐scale identification of proteins. However, 2‐DE is limited by its inability to speed up the in‐gel digestion process. We have developed a new approach to speed up the protein identification process utilizing microwave technology. Proteins excised from gels are subjected to in‐gel digestion with endoprotease trypsin by microwave irradiation, which rapidly produces peptide fragments. The peptide fragments were further analyzed by matrix‐assisted laser desorption/ionization technique for protein identification. The efficacy of this technique for protein mapping was demonstrated by the mass spectral analyses of the peptide fragmentation of several proteins, including lysozyme, albumin, conalbumin, and ribonuclease A. The method reduced the required time for in‐gel digestion of proteins from 16 hours to as little as five minutes. This new application of microwave technology to protein identification will be an important advancement in biotechnology and proteome research.
Automatic annotation of matrix‐assisted laser desorption/ionization N‐glycan spectra
Authors: Goldberg David , Sutton‐Smith Mark , Paulson James , Dell Anne 
Published: February 2005

AbstractMatrix‐assisted laser desorption/ionization‐mass spectrometry (MALDI‐MS) is the pre‐eminent technique for mass mapping of glycans. In order to make this technique practical for high‐throughput screening, reliable automatic methods of annotating peaks must be devised. We describe an algorithm called Cartoonist that labels peaks in MALDI spectra of permethylated N‐glycans with cartoons which represent the most plausible glycans consistent with the peak masses and the types of glycans being analyzed. There are three main parts to Cartoonist. (i) It selects annotations from a library of biosynthetically plausible cartoons. The library we currently use has about 2800 cartoons, but was constructed using only about 300 archetype cartoons entered by hand. (ii) It determines the precision and calibration of the machine used to generate the spectrum. It does this automatically based on the spectrum itself. (iii) It assigns a confidence score to each annotation. In particular, rather than making a binary yes/no decision when annotating a peak, it makes all plausible annotations and associates them with scores indicating the probability that they are correct.
Analysis of O‐glycosylation site occupancy in bovine κ‐casein glycoforms separated by two‐dimensional gel electrophoresis
Authors: Holland John W. , Deeth Hilton C. , Alewood Paul F. 
Published: February 2005

AbstractThe ability of two‐dimensional gel electrophoresis (2‐DE) to separate glycoproteins was exploited to separate distinct glycoforms of κ‐casein that differed only in the number of O‐glycans that were attached. To determine where the glycans were attached, the individual glycoforms were digested in‐gel with pepsin and the released glycopeptides were identified from characteristic sugar ions in the tandem mass spectrometry (MS) spectra. The O‐glycosylation sites were identified by tandem MS after replacement of the glycans with ammonia / aminoethanethiol. The results showed that glycans were not randomly distributed among the five potential glycosylation sites in κ‐casein. Rather, glycosylation of the monoglycoform could only be detected at a single site, T152. Similarly the diglycoform appeared to be modified exclusively at T152 and T163, while the triglycoform was modified at T152, T163 and T154. While low levels of glycosylation at other sites cannot be excluded the hierarchy of site occupation between glycoforms was clearly evident and argues for an ordered addition of glycans to the protein. Since all five potential O‐glycosylation sites can be glycosylated in vivo, it would appear that certain sites remain latent until other sites are occupied. The determination of glycosylation site occupancy in individual glycoforms separated by 2‐DE revealed a distinct pattern of in vivo glycosylation that has not been recognized previously.
Isoelectric beads for proteome pre‐fractionation. II: Experimental evaluation in a multicompartment electrolyzer
Authors: Fortis Frédéric , Girot Pierre , Brieau Olivier , Castagna Annalisa , Righetti Pier Giorgio , Boschetti Egisto 
Published: February 2005

AbstractProteome pre‐fractionation in multicompartment electrolyzers is proposed here, with substantial modifications as compared to the standard technique. First of all, the classical isoelectric, buffering membranes, delimiting each compartment and acting, in pairs, as isoelectric traps, have been replaced by isoelectric buffering beads, operating on the same principle, but allowing unhindered migration of proteins (lack of sieving properties, contrary to typical continuous membrane barriers). Secondly, the isoelectric beads are not made in the conventional manner, with ionic acrylamide derivative monomers throughout their central core, but are composed of a hard, ceramic core, coated with an amphoteric buffering polymer. This minimizes mass transfer resistance of proteins that are transiently adsorbed onto the beads. As a result, significantly reduced separation times, of the order of ca. 3 h, are required for developing steady‐state patterns, as compared to the lengthy times (overnight and much longer) in conventional multicompartment electrolyzers operating with isoelectric membranes. Examples of separation of standard marker proteins, as well as entire Escherichia coli lysates and human serum proteins, are given. The obtained fractions are analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, two‐dimensional gel electrophoresis and by surface enhanced laser desorption/ionization mass spectrometry.
A transcriptomic and proteomic analysis of the effect of CpG‐ODN on human THP‐1 monocytic leukemia cells
Authors: Kuo Cheng‐Chin , Kuo Chu‐Wei , Liang Chi‐Ming , Liang Shu‐Mei 
Published: February 2005

AbstractThe CpG motif of bacterial DNA (CpG‐DNA) is a potent immunostimulating agent whose mechanism of action is not yet clear. Here, we used both DNA microarray and proteomic approaches to investigate the effects of oligodeoxynucleotides containing the CpG motif (CpG‐ODN) on gene transcription and protein expression profiles of CpG‐ODN responsive THP‐1 cells. Microarray analysis revealed that 2 h stimulation with CpG‐ODN up‐regulated 50 genes and down‐regulated five genes. These genes were identified as being associated with inflammation, antimicrobial defense, transcriptional regulation, signal transduction, tumor progression, cell differentiation, proteolysis and metabolism. Longer stimulation (8 h) with CpG‐ODN enhanced transcriptional expression of 58 genes. Among these 58 genes, none except one, namely WNTI inducible signaling pathway protein 2, was the same as those induced after 2 h stimulation. Proteomic analysis by two‐dimensional gel electrophoresis, followed by mass spectrometry identified several proteins up‐regulated by CpG‐ODN. These proteins included heat shock proteins, modulators of inflammation, metabolic proteins and energy pathway proteins. Comparison of microarray and proteomic expression profiles showed poor correlation. Use of more reliable and sensitive analyses, such as reverse transcriptase polymerase chain reaction, Western blotting and functional assays, on several genes and proteins, nonetheless, confirmed that there is indeed good correlation between mRNA and protein expression after CpG‐ODN treatment. This study also revealed that several anti‐apoptotic and neuroprotective related proteins, not previously reported, are activated by CpG‐DNA. These findings have extended our knowledge on the activation of cells by CpG‐DNA and may contribute to further understanding of mechanisms that link innate immunity with acquired immune response(s).
Towards royal jelly proteome
Authors: Scarselli Roberta , Donadio Elena , Giuffrida Maria Gabriella , Fortunato Donatella , Conti Amedeo , Balestreri Ettore , Felicioli Romano , Pinzauti Mauro , Sabatini Anna Gloria , Felicioli Antonio 
Published: February 2005

AbstractThe recent availability of the honey‐bee Apis mellifera genome and trascriptome of both the female castes, has stimulated new efforts in investigating the protein composition of royal jelly (RJ), its role in caste differentiation and its quality and typicality by a proteomic approach. This study is aimed both to separate and identify proteins of royal jelly and to detect some of them in honey‐bee pollen‐bread by using two‐dimensional gel electrophoresis, mass spectrometry and by de novo sequencing. All the identified proteins belonged to the Apis mellifera genome. Apalbumin 1 was also confirmed to be present in honey‐bee pollen‐bread where the presence of apalbumin 2 was also found. In addition several fragments of apalbumin 1 and apalbumin 3 were also found in RJ. These could be the result of protease activity other than that of serine‐protease. This study is a contribution to the description of royal jelly proteome.
Negative‐working photoresist based on a first‐generation dendrimer consisting of 4,4‐diphenylpentyloxy units
Authors: Kamimura Yasushi , Haba Osamu , Endo Takeshi , Ueda Mitsuru 
Published: February 2005

AbstractA first‐generation dendrimer (2), containing phenol groups in the exterior for solubilization in aqueous alkaline solutions, was evaluated as a new negative‐working, alkaline‐developable photoresist material. A negative‐working photoresist based on 2, 4,4′‐methylenebis[2,6‐bis(hydroxymethyl)phenol] as a crosslinker, and diphenyliodonium 9,10‐dimethoxyanthracene‐2‐sulfonate as a photoacid generator was demonstrated. This resist gave a clear negative pattern through postbaking at 90 °C after exposure to UV light, which was followed by development with a 2.38% aqueous tetramethyl ammonium hydroxide solution at room temperature. © 2005 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 43: 1210–1215, 2005