bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comprehensive Analysis of Human Endogenous Retrovirus Transcriptional Activity in Human Tissues with a Retrovirus-Specific Microarray
Authors: Seifarth Wolfgang , Frank Oliver , Zeilfelder Udo , Spiess Birgit , Greenwood Alex D. , Hehlmann Rüdiger , Leib-Mösch Christine 
Published: December 2004

ABSTRACT

Retrovirus-like
sequences account for 8 to 9% of the human genome. Among these
sequences, about 8,000
pol
-containing proviral elements have
been identified to date. As part of our ongoing search for active and
possibly disease-relevant human endogenous retroviruses (HERVs), we
have recently developed an oligonucleotide-based microarray. The assay
allows for both the detection and the identification of most known
retroviral reverse transcriptase (RT)-related nucleic acids in
biological samples. In the present study, we have investigated the
transcriptional activity of representative members of 20 HERV families
in 19 different normal human tissues. Qualitative evaluation of chip
hybridization signals and quantitative analysis by real-time RT-PCR
revealed distinct HERV activity in the human tissues under
investigation, suggesting that HERV elements are active in human cells
in a tissue-specific manner. Most active members of HERV families were
found in mRNA prepared from skin, thyroid gland, placenta, and tissues
of reproductive organs. In contrast, only few active HERVs were
detectable in muscle cells. Human tissues that lack HERV transcription
could not be found, confirming that human endogenous retroviruses are
permanent components of the human transcriptome. Distinct activity
patterns may reflect the characteristics of the regulatory machinery in
these cells, e.g., cell type-dependent occurrence of transcriptional
regulatory
factors.
Kissing-Loop Interaction in the 3′ End of the Hepatitis C Virus Genome Essential for RNA Replication
Authors: Friebe Peter , Boudet Julien , Simorre Jean-Pierre , Bartenschlager Ralf 
Published: December 2004

ABSTRACT

The hepatitis C virus (HCV) is a positive-strand RNA virus belonging to the
Flaviviridae
. Its genome carries at either end highly conserved nontranslated regions (NTRs) containing
cis
-acting RNA elements that are crucial for replication. In this study, we identified a novel RNA element within the NS5B coding sequence that is indispensable for replication. By using secondary structure prediction and nuclear magnetic resonance spectroscopy, we found that this RNA element, designated 5BSL3.2 by analogy to a recent report (S. You, D. D. Stump, A. D. Branch, and C. M. Rice, J. Virol.
78:
1352-1366, 2004), consists of an 8-bp lower and a 6-bp upper stem, an 8-nucleotide-long bulge, and a 12-nucleotide-long upper loop. Mutational disruption of 5BSL3.2 structure blocked RNA replication, which could be restored when an intact copy of this RNA element was inserted into the 3′ NTR. By using this replicon design, we mapped the elements in 5BSL3.2 that are critical for RNA replication. Most importantly, we discovered a nucleotide sequence complementarity between the upper loop of this RNA element and the loop region of stem-loop 2 in the 3′ NTR. Mismatches introduced into the loops inhibited RNA replication, which could be rescued when complementarity was restored. These data provide strong evidence for a pseudoknot structure at the 3′ end of the HCV genome that is essential for replication.
Focal Distribution of Baculovirus IE1 Triggered by Its Binding to the
hr
DNA Elements
Authors: Nagamine Toshihiro , Kawasaki Yu , Iizuka Tetsutaro , Matsumoto Shogo 
Published: December 2004

ABSTRACT

In BmN cells infected with the baculovirus
Bombyx mori
nucleopolyhedrovirus (BmNPV), IE1, a principal transcriptional activator, localizes to sites of viral DNA replication. IE1 initially displays focal distribution in BmNPV-infected cells prior to DNA synthesis, whereas the protein expressed by transfection with the
ie1
gene is distributed throughout the nucleoplasm instead of localized to discrete subnuclear structures. To identify the inducer of focus formation for IE1, we conducted transfection experiments with an
IE1-GFP
construct and found that cotransfection with genomic DNA fragments bearing the homologous region (
hr
) sequences caused the formation of IE1-green fluorescent protein (GFP) foci. The transfection of insect cells with a single plasmid containing exclusively the
hr
3 sequence and the
IE1-GFP
gene was sufficient to form IE1-GFP foci. These results suggest that
hr
elements are a primary determinant of the focal distribution of IE1. An analysis of a series of
hr
3 deletion mutants showed that a single copy of the direct repeat could induce the formation of IE1 foci. Targeted mutagenesis within the
hr
-binding domain of IE1-GFP caused impairment of the
hr
-dependent IE1 localization, suggesting that binding of IE1 to the
hr
elements is essential for the onset of IE1 focus formation. The observation of BmNPV IE1 foci in non-BmNPV-susceptible cells suggests that no species-specific factors are required for
hr
-dependent IE1 focus formation.
CD8+T-Cell-Mediated Cross-Clade Protection in the Genital Tract following Intranasal Immunization with Inactivated Human Immunodeficiency Virus Antigen Plus CpG Oligodeoxynucleotides
Authors: Jiang Janina Q. , Patrick Amy , Moss Ronald B. , Rosenthal Kenneth L. 
Published: December 2004

ABSTRACTHuman immunodeficiency virus (HIV) is a mucosally transmitted infection that rapidly targets and depletes CD4+T cells in mucosal tissues and establishes a major reservoir for viral persistence in gut-associated lymphoid tissues. Therefore, vaccines designed to prevent HIV infections must induce potent and durable mucosal immune responses, especially in the genital tract. Here we investigated whether intranasal (i.n.) immunization with inactivated gp120-depleted HIV-1 antigen (Ag) plus CpG oligodeoxynucleotide (ODN) as an adjuvant induced local immune responses in the genital tract and cross-clade protection against intravaginal (IVAG) challenge. Lymphocytes isolated from the iliac lymph nodes (ILNs) and genital tracts of female mice i.n. immunized with HIV-1 Ag plus CpG showed significant HIV-specific proliferation and produced significantly higher levels of gamma interferon (IFN-γ) and β-chemokines than mice immunized with HIV-1 Ag alone or mixed with non-CpG ODN. CD8+lymphocytes were dramatically increased in the genital tracts of mice immunized with HIV-1 Ag plus CpG, and protection following IVAG challenge with recombinant vaccinia viruses (rVVs) expressing HIV-1 gag was shown to be CD8 dependent. Finally, cross-clade protection was observed between clades A, C, and G but not B following IVAG challenge with rVVs expressing HIV-1gagfrom different clades. These studies provide evidence that mucosal (i.n.) immunization induced strong local T-cell-mediated immune responses in the genital tract and cross-clade protection against IVAG challenge.
Defective Interfering RNA Hinders the Activity of a Tombusvirus-Encoded Posttranscriptional Gene Silencing Suppressor
Authors: Havelda Zoltán , Hornyik Csaba , Válóczi Anna , Burgyán József 
Published: December 2004

ABSTRACT
Defective interfering (DI) RNAs are subviral replicons originating from the viral genome and are associated with many plant RNA viruses and nearly all animal RNA viruses. The presence of DI RNAs in tombusvirus-infected plants reduces the accumulation of helper virus RNA and results in the development of attenuated symptoms similar to those caused by tombusviruses defective in p19, the posttranscriptional gene silencing (PTGS) suppressor. In situ analysis of infected plants containing DI RNAs revealed that the extent of virus infection was spatially restricted as was found for p19-defective tombusvirus. Previously, p19 was shown to suppress PTGS by sequestering the small interfering RNAs (siRNAs), which act as the specificity determinant for PTGS. Our results demonstrate that DI RNAs dramatically elevate the level of virus-specific siRNAs in viral infections, resulting in the saturation of p19 and the accumulation of unbound siRNAs. Moreover, we showed that, at low temperature, where PTGS is inhibited, DI RNAs are not able to efficiently interfere with virus accumulation and protect the plants. These data show that the activation of PTGS plays a pivotal role in DI RNA-mediated interference. Our data also support a role for 21-nucleotide siRNAs in PTGS signaling.
Conditional Cytomegalovirus Replication In Vitro and In Vivo
Authors: Rupp Brigitte , Ruzsics Zsolt , Sacher Torsten , Koszinowski Ulrich H. 
Published: December 2004

ABSTRACT

We have established a conditional gene expression system for cytomegalovirus which allows regulation of genes independently from the viral replication program. Due to the combination of all elements required for regulated expression in the same viral genome, conditional viruses can be studied in different cell lines in vitro and in the natural host in vivo. The combination of a self-sufficient tetracycline-regulated expression cassette and Flp recombinase-mediated insertion into the viral genome allowed fast construction of recombinant murine cytomegaloviruses carrying different conditional genes. The regulation of two reporter genes, the essential viral M50 gene and a dominant-negative mutant gene (
m48.2
) encoding the small capsid protein, was analyzed in more detail. In vitro, viral growth was regulated by the conditional expression of M50 by 3 orders of magnitude and up to a millionfold when the dominant-negative small capsid protein mutant was used. In vivo, viral growth of the dominant-negative mutant was reduced to detection limits in response to the presence of doxycycline in the organs of mice. We believe that this conditional expression system is applicable to genetic studies of large DNA viruses in general.
The Poxviral RING Protein p28 Is a Ubiquitin Ligase That Targets Ubiquitin to Viral Replication Factories
Authors: Nerenberg Bianca T. Hovey , Taylor John , Bartee Eric , Gouveia Kristine , Barry Michele , Früh Klaus 
Published: December 2004

ABSTRACT
The poxviral RING protein p28 is a virulence factor whose molecular function is unknown. Many cellular RING-containing proteins act as ubiquitin ligases (RING-E3s) connecting selected substrate proteins to the ubiquitination machinery. Here we demonstrate that vaccinia virus p28 and its homologue in myxoma virus, M143R, can mediate the formation of polyubiquitin conjugates, while RING mutants of both p28 and M143R cannot. Furthermore, p28 is ubiquitinated in vivo and ubiquitin colocalizes with p28 to virus factories independently of an intact RING domain. These results implicate the ubiquitin system in poxviral virulence.
Long-Term Productive Human Immunodeficiency Virus Infection of CD1a-Sorted Myeloid Dendritic Cells
Authors: Popov Sergei , Chenine Agnès-Laurence , Gruber Andreas , Li Pei-Lin , Ruprecht Ruth M. 
Published: December 2004

ABSTRACTMyeloid, CD1a-sorted dendritic cells (MDC) productively replicated human immunodeficiency virus strains encoding envelope genes of either primary X4R5 or R5 strains for up to 45 days. Cell-free supernatant collected from long-term infected MDC, which had been exposed to an X4R5 virus 45 days earlier, was still infectious when placed over activated T cells. These data imply that DC can act as a persistent reservoir of infectious virus.
Hepatocyte Growth Factor Receptor Is a Coreceptor for Adeno-Associated Virus Type 2 Infection
Authors: Kashiwakura Yuji , Tamayose Kenji , Iwabuchi Kazuhisa , Hirai Yukihiko , Shimada Takashi , Matsumoto Kunio , Nakamura Toshikazu , Watanabe Masami , Oshimi Kazuo , Daida Hiroyuki 
Published: December 2004

ABSTRACT
After the first attachment of virus to the cell surface through a primary receptor, efficient entry of virus requires the presence of a coreceptor. For adeno-associated virus type 2 (AAV2) infection, heparan sulfate proteoglycan is supposed as the primary receptor, and αvβ5 integrin and FGFR1 are reported to act as coreceptors. In this study, we were able to demonstrate that hepatocyte growth factor receptor, c-Met, is also a coreceptor for AAV2 infection. AAV2-mediated transgene analyses revealed that c-Met expression significantly up-regulated transgene expression without increasing AAV2 cell binding. Moreover, a viral overlay assay elucidated the physical interaction between AAV2 and the β subunit of c-Met. These data suggest that c-Met plays the role of coreceptor for AAV2 infection by facilitating AAV2 internalization into the cytoplasm.
Comparative Immunogenicity of Human Immunodeficiency Virus Particles and Corresponding Polypeptides in a DNA Vaccine
Authors: Akahata Wataru , Yang Zhi-yong , Nabel Gary J. 
Published: December 2004

ABSTRACT
The immunogenicity of a plasmid DNA expression vector encoding both Gag and envelope (Env), which produced human immunodeficiency virus (HIV) type 1 virus-like particles (VLP), was compared to vectors expressing Gag and Env individually, which presented the same gene products as polypeptides. Vaccination with plasmids that generated VLP showed cellular immunity comparable to that of Gag and cell-mediated or humoral responses similar to those of Env as immunization with separate vectors. These data suggest that DNA vaccines encoding separated HIV polypeptides generate immune responses similar to those generated by viral particles.
Implementation of team training in medical education in Denmark
Authors: Østergaard H T , Østergaard D , Lippert A 
Published: December 2004

Abstract
In the field of medicine, team training aiming at improving team skills such as leadership, communication, co-operation, and followership at the individual and the team level seems to reduce risk of serious events and therefore increase patient safety. The preferred educational method for this type of training is simulation. Team training is not, however, used routinely in the hospital. In this paper, we describe a framework for the development of a team training course based on need assessment, learning objectives, educational methods including full-scale simulation and evaluations strategies. The use of this framework is illustrated by the present multiprofessional team training in advanced cardiac life support, trauma team training and neonatal resuscitation in Denmark. The challenges of addressing all aspects of team skills, the education of the facilitators, and establishment of evaluation strategies to document the effect of the different types of training on patient safety are discussed.
Alternative service delivery — responding to global pressures
Authors: Fyfe Toby 
Published: December 2004

The new integrated global environment has resulted in a paradigm shift from the nation-state to the international community as a driver of change, an unprecedented degree and speed of change, the replacement of single policy issues with cross-cutting ones and a demand by citizens for improved service delivery. In response, governments are creating alternative service delivery (ASD) arrangements. Jurisdictions using ASD can be placed on a continuum of organizational change. There are slow starters in which all service delivery is state-owned, operated and designed around government needs. There are structural changers that pursue a formulaic approach to organizational change. Finally, there are transformational changers that take a principles-based case-by-case approach to organizational renewal, ensuring that each initiative responds to the particular challenge of the nation and its entities. In Canada, the Policy on Alternative Service Delivery provides guidance for jurisdictions contemplating organizational change with the Public Interest Test. This article will examine the role that organizational change plays in countries that are implementing major governmental change initiatives. It will review the influence of an increasingly integrated global environment on government change and examine how ASD can act as an organizational response to this environment. It will look at the governance issues that countries face when implementing ASD and propose a typology for classifying countries as change agents. Finally, it will reference a principles-based approach for implementing organizational change through ASD that is being implemented in Canada.
Comparative contamination study with cyclophosphamide, fluorouracil and ifosfamide: standard technique versus a proprietary closed-handling system
Authors: Tans Birgit , Willems Ludo 
Published: December 2004

Introduction. In 2003 more than 55 000 preparations of injectable hazardous drugs were made in the central admixture service of the University Hospital in Leuven, Belgium. According to the literature, surface contamination occurs, despite the use of special equipment and procedures. The objective of this study was to compare the surface and glove contamination with cyclophosphamide, ifosfamide and fluorouracil, following preparation using either a standard admixture technique or the fully enclosed PhaSeal® system. Method. During a period of 4 months cyclophosphamide and fluorouracil were prepared with the PhaSeal® system. Following a period of 2 months using the classical technique, cyclophosphamide and ifosfamide were prepared for an 18-month period with the PhaSeal® system. Samples were taken before the start of the 4-month period, in the middle and at the end of the PhaSeal® period, 2 months after stopping PhaSeal® and again after 18 months of working with PhaSeal®. The total number of samples was 104. Results. For fluorouracil only one of the 14 surface samples was detectable, probably because the detection limit is too high. No differences between the results of the surface samples for cyclophosphamide and ifosfamide were detected. However, a rather big spill due to incorrect use of the system could have influenced these results. Differences in contamination on the gloves were more important. Conclusion. Good compliance with standard preparation and cleaning techniques results in equal levels of contamination on surfaces compared with the PhaSeal® system. However, the PhaSeal® system is an important improvement in reducing contamination on gloves.
Pd(II)‐Schiff Base Complexes Heterogenised on MCM‐41 and Delaminated Zeolites as Efficient and Recyclable Catalysts for the Heck Reaction
Authors: González‐Arellano C. , Corma A. , Iglesias M. , Sánchez F. 
Published: December 2004

AbstractWe present a widely applicable approach for the preparation of Pd‐complexes supported on silica‐based mesoporous and laminar inorganic solids (silica gel, purely siliceous and acidic MCM‐41, and delaminated zeolites ITQ‐2 and ITQ‐6) which involves the use of the Schiff bases 2‐tert‐butyl‐4‐methyl‐6{(E)‐[(2S)‐1‐(1‐arylmethyl)pyrrolidinyl]imino}methylphenol (aryl=phenyl, 1‐naphthyl, 2‐naphthyl) as anchored ligands. As an extension of our research in “recyclable catalytic systems” we report results demonstrating the reactivity and recyclability of these systems for the Heck reaction (in a biphasic mode using ethylene glycol and toluene) and Suzuki coupling under phosphine‐free conditions. The complexes are insensitive to oxygen or moisture, and no change of their activity was observed when exposed to an open system during the usual operation. No palladium black was observed after an extended reaction time and no residual palladium was detected from the filtrate at the end of the reaction.
A Novel Thiourea Ligand Applied in the Pd‐Catalyzed Heck, Suzuki and Suzuki Carbonylative Reactions
Authors: Mingji Dai , Liang Bo , Wang Cuihua , You Zejin , Xiang Jing , Dong Guangbin , Chen Jiahua , Yang Zhen 
Published: December 2004

AbstractA novel C2 symmetrical and sterically bulky thiourea ligand 1 has been successfully applied to Heck, Suzuki and Suzuki‐type carbonylative coupling reactions under aerobic conditions. Since the metal‐sulfur bond in the thiourea complexes is stronger than the metal‐phosphorus bond of typical phosphine complexes, thiourea ligands generally do not easily dissociate from the metal center under catalytic conditions, which establishes the thiourea 1‐based palladium complexes as effective catalysts for the palladium‐catalyzed cross‐coupling reactions.
Arylation of Carbonyl Compounds Catalyzed by Rhodium and Iridium 1,3‐R2‐Tetrahydropyrimidin‐2‐ylidenes: Structure‐Reactivity Correlations
Authors: Imlinger Nicolas , Mayr Monika , Wang Dongren , Wurst Klaus , Buchmeiser Michael R. 
Published: December 2004

AbstractSix different well‐defined rhodium and iridium N‐heterocyclic carbene complexes, i.e., RhCl(1,3‐dimesityltetrahydropyrimidin‐2‐ylidene)(COD) (1), RhBr(1,3‐dimesityltetrahydropyrimidin‐2‐ylidene)(COD) (2), RhCl[1,3‐di(2‐propyl)tetrahydropyrimidin‐2‐ylidene](COD) (3), IrCl(1,3‐dimesityltetrahydropyrimidin‐2‐ylidene)(COD) (4), Rh(CF3COO)(1,3‐dimesityltetrahydropyrimidin‐2‐ylidene)(COD) (5), and IrBr[1,3‐di(2‐propyl)tetrahydropyrimidin‐2‐ylidene](COD) (6) (COD=1,5‐cyclooctadiene, mesityl=2,4,6‐trimethylphenyl) have been used as catalysts for the arylation of aldehydes and α,β‐unsaturated ketones using different arylboronic acids. Compounds 1–4 and 6 were prepared by reaction of [RhCl(COD)]2 and [IrCl(COD)]2, respectively, with a base and the corresponding 1,3‐R2‐tetrahydropyrimidinium salt. Compound 5 was prepared by reaction of 1.0 equivalents of CF3COOAg with 1. The use of an excess of CF3COOAg resulted in the replacement of Rh(I) by Ag(I) and yielded Ag(1,3‐dimesityltetrahydropyrimidin‐2‐ylidene)+Rh2(CF3COO)3(COD)− (8). Compounds 4 and 8 were characterized by X‐ray analysis. The activity of the rhodium complexes increased in the order 5>3>1>2, indicating the necessity of strongly electron‐withdrawing groups at the metal centers, thus increasing their nucleophilicity. In due consequence, the “softer” iridium complexes 4 and 6 exhibited significantly reduced catalytic activity albeit with enhanced selectivity. The syntheses of the metal complexes as well as a detailed study on their reactivity in the arylation of carbonyl compounds using equimolar amounts of arylboronic acid and carbonyl compound in the presence of 0.08–1 mol % catalyst are presented.
Aromatic CH Borylation Catalyzed by Iridium/2,6‐Diisopropyl‐N‐(2‐pyridylmethylene)aniline Complex
Authors: Tagata Tsuyosi , Nishida Mayumi 
Published: December 2004

AbstractThe CH borylation of aromatic and heteroaromatic compounds, such as benzene, 1,3‐dichlorobenzene, 1,3‐bis(trifluoromethyl)benzene, 2,6‐dichloropyridine, indole, benzothiophene and benzofuran, by bis(pinacolato)diboron or pinacolborane was catalyzed by a 1/2 [IrCl(COD)]2‐2,6‐diisopropyl‐N‐(2‐pyridylmethylene)aniline complex. The isopropyl groups of the ligand are essential for obtaining the products in high yield. Octane was a suitable solvent for the reactions of the above compounds except for indole. In the case of indole, DME was better than octane and the yield tended to improve with a smaller amount of the catalyst.
Novel Nickel‐Catalyzed Coupling Reaction of Allyl Ethers with Chlorosilanes, Alkyl Tosylates, or Alkyl Halides Promoted by Vinyl‐Grignard Reagent Leading to Allylsilanes or Alkenes
Authors: Terao Jun , Watabe Hiroyasu , Watanabe Hiroyuki , Kambe Nobuaki 
Published: December 2004

AbstractA new method for a carbon‐silicon or carbon‐carbon bond forming reaction between allyl ethers and chlorosilanes, alkyl tosylates, or alkyl halides giving rise to allylsilanes or alkenes has been developed. This reaction proceeds efficiently at ambient temperature by the combined use of nickel catalysts and a vinyl‐Grignard reagent. A possible reaction pathway involving the formation of allyl‐Grignard reagents via transmetallation of π‐allylnickel complexes with the vinyl‐Grignard reagent and subsequent trapping of the thus formed allyl‐Grignard reagents with electrophiles is proposed.